Pub Date : 2024-12-01DOI: 10.19540/j.cnki.cjcmm.20240902.705
Hui-Yu Zhang, Gao-Jie Xin, Yuan-Yuan Chen, Ce Cao, Xiao-Shan Cui, Jia-Ming Gao, Hao Guo, Jian-Hua Fu
This article explored the specific mechanism by which ginsenoside Rg_1 regulates cellular autophagy to attenuate hypoxia/reoxygenation(H/R) injury in HL-1 cardiomyocytes through the microRNA155(miR-155)/neurogenic gene Notch homologous protein 1(Notch1)/hairy and enhancer of split 1(Hes1) pathway. An HL-1 cell model with H/R injury was constructed, and ginsenoside Rg_1 and/or Notch1 inhibitor DAPT and miR-155 mimics were used to treat cells. Cell counting kit(CCK)-8 was used to detect the relative viability of HL-1 cells with H/R injury. The lactate dehydrogenase(LDH) content in cell culture medium supernatant was detected by using an LDH assay kit, and autophagosome in cells was observed by transmission electron microscopy. The level of autophagy in cells was detected through the mono-dansyl-cadaverine(MDC) detection method. Fluorescence quantitative polymerase chain reaction was used to detect the mRNA levels of miR-155, Notch1, Hes1, and microtubule-associated protein1 light chain 3(LC3), and Western blot was used to detect the protein expression levels of Notch1, Hes1, LC3Ⅰ, and LC3Ⅱ. The results show that after H/R injury, the activity of HL-1 cells decreases, and LDH leakage increases. Besides, the number of intracellular autophagosomes increases, and the mRNA level of LC3 and the LC3Ⅱ/LC3Ⅰ ratio are elevated. In addition, ginsenoside Rg_1 can increase cell activity, decrease LDH leakage and the number of intracellular autophagosomes, and reduce the mRNA level of LC3 and the LC3Ⅱ/LC3Ⅰ ratio. Therefore, it plays a cardioprotective role by inhibiting autophagy, and Notch1 inhibitor or miR-155 overexpression can inhibit the effect of ginsenoside Rg_1, promote autophagy, and aggravate H/R injury in HL-1 cells. Ginsenoside Rg_1 can inhibit the reduction of Notch1 and Hes1 mRNA levels and protein expressions and the increase in miR-155 mRNA levels caused by H/R injury, while Notch1 inhibitors or miR-155 overexpression show the opposite effect. In summary, ginsenoside Rg_1 can regulate autophagy through the miR-155/Notch1/Hes1 pathway to alleviate H/R injury in HL-1 cardiomyocytes.
本文探讨了人参皂苷Rg_1通过microRNA155(miR-155)/神经源性基因Notch同源蛋白1(Notch1)/hairy and enhancer of split 1(Hes1)通路调控细胞自噬减轻HL-1心肌细胞缺氧/再氧化(H/R)损伤的具体机制。构建H/R损伤的HL-1细胞模型,用人参皂苷Rg_1和/或Notch1抑制剂DAPT和miR-155模拟物处理细胞。采用细胞计数试剂盒(CCK)-8检测H/R损伤后HL-1细胞的相对活力。采用乳酸脱氢酶(LDH)检测试剂盒检测细胞培养基上清中乳酸脱氢酶(LDH)的含量,透射电镜观察细胞自噬体的变化。采用单丹酰尸胺(MDC)检测法检测细胞自噬水平。采用荧光定量聚合酶链式反应检测miR-155、Notch1、Hes1、微管相关蛋白1轻链3(LC3) mRNA表达水平,采用Western blot检测Notch1、Hes1、LC3Ⅰ、LC3Ⅱ蛋白表达水平。结果表明,H/R损伤后,HL-1细胞活性降低,LDH渗漏增加。细胞内自噬体数量增加,LC3 mRNA水平升高,LC3Ⅱ/LC3Ⅰ比值升高。人参皂苷Rg_1可以提高细胞活性,减少LDH渗漏和细胞内自噬体数量,降低LC3 mRNA水平和LC3Ⅱ/LC3Ⅰ比值。因此,它通过抑制自噬发挥心脏保护作用,Notch1抑制剂或miR-155过表达可抑制人参皂苷Rg_1的作用,促进自噬,加重HL-1细胞的H/R损伤。人参皂苷Rg_1可以抑制H/R损伤引起的Notch1和Hes1 mRNA水平和蛋白表达的降低以及miR-155 mRNA水平的升高,而Notch1抑制剂或miR-155过表达则相反。综上所述,人参皂苷Rg_1可以通过miR-155/Notch1/Hes1通路调节自噬,减轻HL-1心肌细胞H/R损伤。
{"title":"[Mechanism of ginsenoside Rg_1 in regulating autophagy through miR-155/Notch1/Hes1 pathway to attenuate hypoxia/reoxygenation injury in HL-1 cells].","authors":"Hui-Yu Zhang, Gao-Jie Xin, Yuan-Yuan Chen, Ce Cao, Xiao-Shan Cui, Jia-Ming Gao, Hao Guo, Jian-Hua Fu","doi":"10.19540/j.cnki.cjcmm.20240902.705","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240902.705","url":null,"abstract":"<p><p>This article explored the specific mechanism by which ginsenoside Rg_1 regulates cellular autophagy to attenuate hypoxia/reoxygenation(H/R) injury in HL-1 cardiomyocytes through the microRNA155(miR-155)/neurogenic gene Notch homologous protein 1(Notch1)/hairy and enhancer of split 1(Hes1) pathway. An HL-1 cell model with H/R injury was constructed, and ginsenoside Rg_1 and/or Notch1 inhibitor DAPT and miR-155 mimics were used to treat cells. Cell counting kit(CCK)-8 was used to detect the relative viability of HL-1 cells with H/R injury. The lactate dehydrogenase(LDH) content in cell culture medium supernatant was detected by using an LDH assay kit, and autophagosome in cells was observed by transmission electron microscopy. The level of autophagy in cells was detected through the mono-dansyl-cadaverine(MDC) detection method. Fluorescence quantitative polymerase chain reaction was used to detect the mRNA levels of miR-155, Notch1, Hes1, and microtubule-associated protein1 light chain 3(LC3), and Western blot was used to detect the protein expression levels of Notch1, Hes1, LC3Ⅰ, and LC3Ⅱ. The results show that after H/R injury, the activity of HL-1 cells decreases, and LDH leakage increases. Besides, the number of intracellular autophagosomes increases, and the mRNA level of LC3 and the LC3Ⅱ/LC3Ⅰ ratio are elevated. In addition, ginsenoside Rg_1 can increase cell activity, decrease LDH leakage and the number of intracellular autophagosomes, and reduce the mRNA level of LC3 and the LC3Ⅱ/LC3Ⅰ ratio. Therefore, it plays a cardioprotective role by inhibiting autophagy, and Notch1 inhibitor or miR-155 overexpression can inhibit the effect of ginsenoside Rg_1, promote autophagy, and aggravate H/R injury in HL-1 cells. Ginsenoside Rg_1 can inhibit the reduction of Notch1 and Hes1 mRNA levels and protein expressions and the increase in miR-155 mRNA levels caused by H/R injury, while Notch1 inhibitors or miR-155 overexpression show the opposite effect. In summary, ginsenoside Rg_1 can regulate autophagy through the miR-155/Notch1/Hes1 pathway to alleviate H/R injury in HL-1 cardiomyocytes.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6450-6458"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to rapidly identify the chemical components in Dracocephalum moldavica, and UPLC was employed to determine the content of its main components. MS analysis was performed using an electrospray ionization(ESI) source and data were collected in the negative ion mode. By comparing the retention time and mass spectra of reference compounds, and using a self-built compound database and the PubChem database, 68 compounds were identified from D. moldavica, including 36 flavonoids, 22 phenylpropanoids, 4 phenols, and 6 other compounds. On this basis, a UPLC quantitative method was established to simultaneously determine 8 main components, i.e., luteolin-7-O-glucuronide, apigenin-7-O-glucuronide, rosmarinic acid, diosmetin-7-O-glucuronide, tilianin, acacetin-7-O-glucuronide, acacetin-7-O-(6″-O-malonyl)-glucoside, and acacetin. A Waters ACQUITY BEH C_(18) column(2.1 mm × 100 mm, 1.7 μm) was used, with acetonitrile and a water solution containing 0.1% formic acid and 0.1% phosphoric acid as the mobile phase for gradient elution. The detection wavelength was set at 330 nm, with a flow rate of 0.4 mL·min~(-1), and the column temperature was maintained at 35 ℃. The 8 components demonstrated good linearity(r≥0.999 9) over a wide mass concentration range(50 or 100 times). The average recovery rate ranged from 97.5% to 105.1%, and the relative standard deviations(RSDs) were 0.90% to 3.4%(n= 6), indicating that the method was simple, accurate, and reliable. In 17 batches of D. moldavica samples, the content of these 8 components ranged from 0.405 to 2.10, 0.063 to 0.342, 0.446 to 2.43, 0.415 to 1.47, 1.57 to 4.34, 0.173 to 0.386, 1.00 to 5.40, and 0.069 to 0.207 mg·g~(-1), respectively. These results indicate significant differences in the internal quality of the samples, highlighting the need for strict quality control to ensure their pharmacodynamic efficacy. This study provides a scientific basis for the rapid discovery of pharmacodynamic substances, comprehensive quality control, and the formulation or revision of quality standards for D. moldavica.
采用超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC- q - tof -MS/MS)对龙头草中的化学成分进行快速鉴定,并采用超高效液相色谱法测定其主要成分的含量。质谱分析采用电喷雾电离(ESI)源,在负离子模式下采集数据。通过比较参比化合物的保留时间和质谱,并结合自建的化合物数据库和PubChem数据库,从moldavica中鉴定出68种化合物,其中黄酮类化合物36种,苯丙类化合物22种,酚类化合物4种,其他化合物6种。在此基础上,建立了UPLC定量方法,同时测定木犀草素-7- o -葡糖苷、芹菜素-7- o -葡糖苷、迭香香酸、薯蓣皂苷-7- o -葡糖苷、天青素、棘球葡糖苷-7- o -葡糖苷(6″- o -丙二醇基)、棘球葡糖苷等8种主要成分。采用Waters ACQUITY BEH C_(18)色谱柱(2.1 mm × 100 mm, 1.7 μm),以乙腈和0.1%甲酸、0.1%磷酸水溶液为流动相梯度洗脱。检测波长为330 nm,流速为0.4 mL·min~(-1),柱温为35℃。在较宽的质量浓度范围内(50倍或100倍),8种成分均表现出良好的线性关系(r≥0.999 9)。平均加样回收率为97.5% ~ 105.1%,相对标准偏差(rsd)为0.90% ~ 3.4%(n= 6),方法简便、准确、可靠。17批moldavica样品中8种成分的含量分别为0.405 ~ 2.10、0.063 ~ 0.342、0.446 ~ 2.43、0.415 ~ 1.47、1.57 ~ 4.34、0.173 ~ 0.386、1.00 ~ 5.40和0.069 ~ 0.207 mg·g~(-1)。这些结果表明,样品的内部质量存在显著差异,强调需要严格的质量控制,以确保其药效学效果。本研究为快速发现药效学物质、进行全面的质量控制以及制定或修订石竹药材质量标准提供了科学依据。
{"title":"[Qualitative and quantitative analysis of chemical components of Dracocephalum moldavica based on UPLC-Q-TOF-MS/MS and UPLC].","authors":"Ming-Lei Xu, Hui-Min Gao, Yong-Xin Zhang, Zhi-Jian Li, Yang Ding, Qing-Rong Wang, Shi-Xia Huo, Wei-Hong Feng, Yu-Tong Kang, Liang-Mian Chen, Zhi-Min Wang","doi":"10.19540/j.cnki.cjcmm.20240910.301","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240910.301","url":null,"abstract":"<p><p>Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to rapidly identify the chemical components in Dracocephalum moldavica, and UPLC was employed to determine the content of its main components. MS analysis was performed using an electrospray ionization(ESI) source and data were collected in the negative ion mode. By comparing the retention time and mass spectra of reference compounds, and using a self-built compound database and the PubChem database, 68 compounds were identified from D. moldavica, including 36 flavonoids, 22 phenylpropanoids, 4 phenols, and 6 other compounds. On this basis, a UPLC quantitative method was established to simultaneously determine 8 main components, i.e., luteolin-7-O-glucuronide, apigenin-7-O-glucuronide, rosmarinic acid, diosmetin-7-O-glucuronide, tilianin, acacetin-7-O-glucuronide, acacetin-7-O-(6″-O-malonyl)-glucoside, and acacetin. A Waters ACQUITY BEH C_(18) column(2.1 mm × 100 mm, 1.7 μm) was used, with acetonitrile and a water solution containing 0.1% formic acid and 0.1% phosphoric acid as the mobile phase for gradient elution. The detection wavelength was set at 330 nm, with a flow rate of 0.4 mL·min~(-1), and the column temperature was maintained at 35 ℃. The 8 components demonstrated good linearity(r≥0.999 9) over a wide mass concentration range(50 or 100 times). The average recovery rate ranged from 97.5% to 105.1%, and the relative standard deviations(RSDs) were 0.90% to 3.4%(n= 6), indicating that the method was simple, accurate, and reliable. In 17 batches of D. moldavica samples, the content of these 8 components ranged from 0.405 to 2.10, 0.063 to 0.342, 0.446 to 2.43, 0.415 to 1.47, 1.57 to 4.34, 0.173 to 0.386, 1.00 to 5.40, and 0.069 to 0.207 mg·g~(-1), respectively. These results indicate significant differences in the internal quality of the samples, highlighting the need for strict quality control to ensure their pharmacodynamic efficacy. This study provides a scientific basis for the rapid discovery of pharmacodynamic substances, comprehensive quality control, and the formulation or revision of quality standards for D. moldavica.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6352-6367"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Identification of critical material attributes(CMAs) is a key issue in the quality control of large-scale TCM products like Jianwei Xiaoshi Tablets. This study focuses on the granules of Jianwei Xiaoshi Tablets, using tablet tensile strength as the primary quality attribute. A method for identifying the CMAs and a design space for the granules were established, along with a predictive model for the granule CMAs based on Fourier transform near-infrared spectroscopy(FT-NIR). First, granules of Jianwei Xiaoshi Tablets with different properties were prepared using a partial factorial design method from the design of experiments(DOE). The powder properties of the granules were measured. An orthogonal partial least squares(OPLS) model was established to correlate the powder properties with tensile strength. Based on the characteristics of the comprehensive variables extracted by OPLS, the independent variables with the greatest explanatory power for tensile strength were identified. FT-NIR technology was then employed to establish a predictive model for the granule CMAs. The final CMAs identified were hygroscopicity, moisture content, D_(50), collapse angle, mass flow rate, and tapped density. The coefficients of determination of the prediction set(R■) and relative percentage deviation(RPD) of the prediction set for flowability, D_(50), and moisture content were 0.891, 0.994, and 0.998; and 2.97, 12.4, and 20.7, respectively. The established OPLS model clearly identified the impact of various factors on tensile strength, demonstrating good fit results. The model exhibited high prediction accuracy and can be used for the rapid and accurate determination of CMAs in granules of Jianwei Xiaoshi Tablets.
{"title":"[Identification of CMAs of Jianwei Xiaoshi Tablet granules based on QbD concept and construction of their predictive model].","authors":"Xin-Hao Wan, Zhi-Jian Zhong, Qing Tao, Zi-Qian Wang, Jia-Li Liao, Dong-Yin Yang, Ming Yang, Xiao-Rong Luo, Zhen-Feng Wu","doi":"10.19540/j.cnki.cjcmm.20241022.301","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20241022.301","url":null,"abstract":"<p><p>Identification of critical material attributes(CMAs) is a key issue in the quality control of large-scale TCM products like Jianwei Xiaoshi Tablets. This study focuses on the granules of Jianwei Xiaoshi Tablets, using tablet tensile strength as the primary quality attribute. A method for identifying the CMAs and a design space for the granules were established, along with a predictive model for the granule CMAs based on Fourier transform near-infrared spectroscopy(FT-NIR). First, granules of Jianwei Xiaoshi Tablets with different properties were prepared using a partial factorial design method from the design of experiments(DOE). The powder properties of the granules were measured. An orthogonal partial least squares(OPLS) model was established to correlate the powder properties with tensile strength. Based on the characteristics of the comprehensive variables extracted by OPLS, the independent variables with the greatest explanatory power for tensile strength were identified. FT-NIR technology was then employed to establish a predictive model for the granule CMAs. The final CMAs identified were hygroscopicity, moisture content, D_(50), collapse angle, mass flow rate, and tapped density. The coefficients of determination of the prediction set(R■) and relative percentage deviation(RPD) of the prediction set for flowability, D_(50), and moisture content were 0.891, 0.994, and 0.998; and 2.97, 12.4, and 20.7, respectively. The established OPLS model clearly identified the impact of various factors on tensile strength, demonstrating good fit results. The model exhibited high prediction accuracy and can be used for the rapid and accurate determination of CMAs in granules of Jianwei Xiaoshi Tablets.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 24","pages":"6565-6573"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-01DOI: 10.19540/j.cnki.cjcmm.20240904.402
Feng Chen, Nai-Fan Duan, Xing Zhang, Wei Zhang
To investigate the therapeutic effect of Fuzheng Tongluo Granules on idiopathic pulmonary fibrosis(IPF) and its mechanism. Seventy-two SD rats were randomly divided into the control group, model group, pirfenidone group(162 mg·kg~(-1)), and low-, medium-and high-dose of Fuzheng Tongluo Granules groups(2.63, 5.25, 10.5 g·kg~(-1)). Rat model of IPF was induced by a single non-invasive tracheal intubation drip of bleomycin(BLM). The corresponding drugs were given daily by gavage after the 2nd day of modeling, and body mass was recorded. On the 28th day, the samples were collected and weighed, and the lung coefficients were calculated. The pathological changes in the lung tissue were observed by HE and Masson staining, and the hydroxyproline(HYP) content of the lung tissue was detected by alkaline hydrolysis. The contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-18(IL-18) of the lung tissue were determined by ELISA. The expression of collagen type Ⅰ(collagen Ⅰ) and α-smooth muscle actin(α-SMA) was observed by immunohistochemistry. The expression levels of NOD-, LRR-and pyrin domain-containing 3(NLRP3), cysteine-requiring aspartate protease type 1(caspase-1), gasdermin D-N(GSDMD-N), and apoptosis-associated speck-like protein containing a CARD(ASC) in the lung tissue were detected by Western blot. Immunofluorescence co-localization was used to observe the expression of GSDMD and CD68. The results show that compared with the control group, the model group showed increased lung coefficient, Ashcroft score, Szapiel score, HYP, TNF-α, IL-1β, and IL-18 content in the lung tissue and elevated protein expression levels of NLRP3, caspase-1, GSDMD-N, and ASC. The expression levels of GSDMD and CD68 were increased, and there was a high degree of co-localization between GSDMD and CD68. Compared with those in the model group, the lung coefficient, Ashcroft score, and Szapiel score decreased in all drug administration groups, and the content of HYP, TNF-α, IL-1β, and IL-18 decreased. The protein expression levels of NLRP3, caspase-1, GSDMD-N, and ASC decreased, and the expression levels of GSDMD and CD68 were reduced. There was a high degree of co-localization between GSDMD and CD68. In summary, Fuzheng Tongluo Granules can effectively reduce pulmonary fibrosis and inflammation levels in rats with IPF, and the mechanism may be related to the down-regulation of the NLRP3/caspase-1/GSDMD pathway to inhibit macrophage pyroptosis.
{"title":"[Effect of Fuzheng Tongluo Granules on macrophage pyroptosis in rat model with pulmonary fibrosis based on NLRP3/caspase-1/GSDMD pathway].","authors":"Feng Chen, Nai-Fan Duan, Xing Zhang, Wei Zhang","doi":"10.19540/j.cnki.cjcmm.20240904.402","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240904.402","url":null,"abstract":"<p><p>To investigate the therapeutic effect of Fuzheng Tongluo Granules on idiopathic pulmonary fibrosis(IPF) and its mechanism. Seventy-two SD rats were randomly divided into the control group, model group, pirfenidone group(162 mg·kg~(-1)), and low-, medium-and high-dose of Fuzheng Tongluo Granules groups(2.63, 5.25, 10.5 g·kg~(-1)). Rat model of IPF was induced by a single non-invasive tracheal intubation drip of bleomycin(BLM). The corresponding drugs were given daily by gavage after the 2nd day of modeling, and body mass was recorded. On the 28th day, the samples were collected and weighed, and the lung coefficients were calculated. The pathological changes in the lung tissue were observed by HE and Masson staining, and the hydroxyproline(HYP) content of the lung tissue was detected by alkaline hydrolysis. The contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-18(IL-18) of the lung tissue were determined by ELISA. The expression of collagen type Ⅰ(collagen Ⅰ) and α-smooth muscle actin(α-SMA) was observed by immunohistochemistry. The expression levels of NOD-, LRR-and pyrin domain-containing 3(NLRP3), cysteine-requiring aspartate protease type 1(caspase-1), gasdermin D-N(GSDMD-N), and apoptosis-associated speck-like protein containing a CARD(ASC) in the lung tissue were detected by Western blot. Immunofluorescence co-localization was used to observe the expression of GSDMD and CD68. The results show that compared with the control group, the model group showed increased lung coefficient, Ashcroft score, Szapiel score, HYP, TNF-α, IL-1β, and IL-18 content in the lung tissue and elevated protein expression levels of NLRP3, caspase-1, GSDMD-N, and ASC. The expression levels of GSDMD and CD68 were increased, and there was a high degree of co-localization between GSDMD and CD68. Compared with those in the model group, the lung coefficient, Ashcroft score, and Szapiel score decreased in all drug administration groups, and the content of HYP, TNF-α, IL-1β, and IL-18 decreased. The protein expression levels of NLRP3, caspase-1, GSDMD-N, and ASC decreased, and the expression levels of GSDMD and CD68 were reduced. There was a high degree of co-localization between GSDMD and CD68. In summary, Fuzheng Tongluo Granules can effectively reduce pulmonary fibrosis and inflammation levels in rats with IPF, and the mechanism may be related to the down-regulation of the NLRP3/caspase-1/GSDMD pathway to inhibit macrophage pyroptosis.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6399-6406"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143056140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study systematically reviewed the literature on the prevention and treatment of colorectal cancer(CRC) with traditional Chinese medicine(TCM), aiming to present a more intuitive and concise overview of existing evidence. Four major Chinese databases, including CNKI, Wanfang, VIP, and SinoMed, were searched for randomized controlled trial(RCT) on TCM treatment of CRC. The retrieval period was from database inception to August 1, 2023. The evidence was presented using a combination of text and charts. A total of 1 778 RCTs were included, and the overall publication volume showed an upward trend. The quality of the RCT was generally low, with sample sizes concentrated between 60 and 100 cases. The intervention durations were mainly 4, 8, and 12 weeks. Keywords primarily focused on advanced CRC, postoperative CRC, immune function, and gastrointestinal function. Clinical complications were often caused by surgery or chemotherapy, including intestinal obstruction, peripheral neuropathy, diarrhea, and anxiety and depression. There were various intervention measures, including TCM decoctions, TCM injections, Chinese medicine nursing, Chinese patent medicines, and acupuncture. Among them, TCM decoctions(excluding self-made prescriptions) included Shenling Baizhu Powder(32 articles, 1.80%) and Sijunzi Decoction(22 articles, 1.24%). TCM injections included Fufang Kushen Injection(54 articles, 3.04%) and Aidi Injection(46 articles, 2.59%). Chinese patent medicines included Cinobufacin Capsules(16 articles, 0.90%) and Fufang Banmao Capsules(10 articles, 0.56%). The outcome indicators were divided into 13 domains, including recent efficacy, quality of life, safety events, and TCM syndrome/symptom scores. The existing outcome indicators mostly followed the western medicine evaluation system, with complex types and no unified standards, lacking outcome indicators or scales with TCM characteristics, and relatively insufficient attention to long-term efficacy, anxiety, and depression. Future research should optimize clinical study designs, build a core index set and clinical evaluation system with TCM characteristics, and produce more high-level evidence to support the safety and effectiveness of TCM in preventing and treating CRC.
{"title":"[Evidence map for randomized controlled trials of traditional Chinese medicine in prevention and treatment of colorectal cancer].","authors":"Dong Zhang, Ming-Xin Ni, Xiao-Man Wei, Xue-Chen Geng, Liu Li, Hai-Bo Cheng","doi":"10.19540/j.cnki.cjcmm.20240812.501","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240812.501","url":null,"abstract":"<p><p>This study systematically reviewed the literature on the prevention and treatment of colorectal cancer(CRC) with traditional Chinese medicine(TCM), aiming to present a more intuitive and concise overview of existing evidence. Four major Chinese databases, including CNKI, Wanfang, VIP, and SinoMed, were searched for randomized controlled trial(RCT) on TCM treatment of CRC. The retrieval period was from database inception to August 1, 2023. The evidence was presented using a combination of text and charts. A total of 1 778 RCTs were included, and the overall publication volume showed an upward trend. The quality of the RCT was generally low, with sample sizes concentrated between 60 and 100 cases. The intervention durations were mainly 4, 8, and 12 weeks. Keywords primarily focused on advanced CRC, postoperative CRC, immune function, and gastrointestinal function. Clinical complications were often caused by surgery or chemotherapy, including intestinal obstruction, peripheral neuropathy, diarrhea, and anxiety and depression. There were various intervention measures, including TCM decoctions, TCM injections, Chinese medicine nursing, Chinese patent medicines, and acupuncture. Among them, TCM decoctions(excluding self-made prescriptions) included Shenling Baizhu Powder(32 articles, 1.80%) and Sijunzi Decoction(22 articles, 1.24%). TCM injections included Fufang Kushen Injection(54 articles, 3.04%) and Aidi Injection(46 articles, 2.59%). Chinese patent medicines included Cinobufacin Capsules(16 articles, 0.90%) and Fufang Banmao Capsules(10 articles, 0.56%). The outcome indicators were divided into 13 domains, including recent efficacy, quality of life, safety events, and TCM syndrome/symptom scores. The existing outcome indicators mostly followed the western medicine evaluation system, with complex types and no unified standards, lacking outcome indicators or scales with TCM characteristics, and relatively insufficient attention to long-term efficacy, anxiety, and depression. Future research should optimize clinical study designs, build a core index set and clinical evaluation system with TCM characteristics, and produce more high-level evidence to support the safety and effectiveness of TCM in preventing and treating CRC.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6512-6520"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-01DOI: 10.19540/j.cnki.cjcmm.20240912.702
Chun-Lin Wu, Ya-Nan Hu, Yi-Qiang Liu, Hui Li, Quan Wen
<p><p>The aim of this study was to investigate the underlying mechanism of chrysophanol(Chr) in reducing inflammation and foam cell formation induced by oxidized low-density lipoprotein(ox-LDL) and to investigate the targets and pathways related to effects of Chr on coronary atherosclerosis, providing a theoretical basis for the development of new clinical drugs. RAW264.7 macrophages were cultured in vitro, and after determining the appropriate concentrations of Chr and ox-LDL for treating RAW264.7 macrophages using a cell counting kit-8(CCK-8), the macrophages were treated with different concentrations of Chr(10, 15 μmol·L~(-1)) and ox-LDL(with or without 80 mg·mL~(-1)) for 24 h. RAW264.7 macrophages were divided into four groups: control group, model group(80 mg·mL~(-1) ox-LDL), treatment group(80 mg·mL~(-1) ox-LDL+10 μmol·L~(-1) Chr), and treatment group(80 mg·mL~(-1) ox-LDL+15 μmol·L~(-1) Chr). Lipid accumulation in each group was detected by oil red O staining. CD36 expression was analyzed by flow cytometry. Western blot was used to detect the expression of scavenger receptor class A1(SR-A1), scavenger receptor class B type Ⅰ(SR-B1), autophagy-related protein 5(Atg5), Beclin-1, autophagy adaptor protein p62(P62), the ratio of microtubule-associated protein light chain 3(LC3)Ⅱ to LC3Ⅰ(LC3Ⅱ/LC3Ⅰ), nuclear factor kappa B P65(NF-κB P65), inhibitor of κB kinase β(IKKβ), nuclear factor of κB inhibitor(IκB), high mobility group box protein 1(HMGB1), phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), and phosphorylated mammalian target of rapamycin(mTOR). Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression levels of ATP-binding cassette transporter A1(ABCA1), ATP-binding cassette transporter G1(ABCG1), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), HMGB1, inducible nitric oxide synthase(iNOS), arginase 1(Arg1), macrophage galactose-type lectin-1(Mgl-1), and NF-κB P65. Immunofluorescence analysis was performed to determine the localization of HMGB1 in RAW264.7 cells in each group. The autophagy inhibitor 3-methyladenine(3-MA) was added as a control for reverse validation, and the RAW264.7 macrophages were divided into four groups again: control group, model group(80 mg·mL~(-1) ox-LDL), treatment group(80 mg·mL~(-1) ox-LDL + 15 μmol·L~(-1) Chr), and inhibitor group(80 mg·mL~(-1) ox-LDL+15 μmol·L~(-1) Chr+3-MA). The results showed that Chr effectively reduced foam cell formation by regulating the expression levels of SR-A1, ABCA1, ABCG1, the LC3Ⅱ/LC3Ⅰ ratio, Atg5, Beclin-1, and p62, and inhibited the NF-κB/HMGB1-PI3K/Akt/mTOR signaling pathway. Moreover, the inhibitory effects of Chr on autophagy and the NF-κB/HMGB1-PI3K/Akt/mTOR pathway were reversed by the autophagy inhibitor 3-MA. In conclusion, Chr exhibits therapeutic potential for the treatment of atherosclerosis by inducing autophagy and modulating the NF-κB/HMGB1 and PI3K/Akt/mTOR pathways to inhibit the formation of macrophage infla
{"title":"[Mechanism of chrysophanol in inhibiting ox-LDL-induced macrophage foaminess through NF-κB/HMGB1-PI3K/Akt/mTOR pathway].","authors":"Chun-Lin Wu, Ya-Nan Hu, Yi-Qiang Liu, Hui Li, Quan Wen","doi":"10.19540/j.cnki.cjcmm.20240912.702","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240912.702","url":null,"abstract":"<p><p>The aim of this study was to investigate the underlying mechanism of chrysophanol(Chr) in reducing inflammation and foam cell formation induced by oxidized low-density lipoprotein(ox-LDL) and to investigate the targets and pathways related to effects of Chr on coronary atherosclerosis, providing a theoretical basis for the development of new clinical drugs. RAW264.7 macrophages were cultured in vitro, and after determining the appropriate concentrations of Chr and ox-LDL for treating RAW264.7 macrophages using a cell counting kit-8(CCK-8), the macrophages were treated with different concentrations of Chr(10, 15 μmol·L~(-1)) and ox-LDL(with or without 80 mg·mL~(-1)) for 24 h. RAW264.7 macrophages were divided into four groups: control group, model group(80 mg·mL~(-1) ox-LDL), treatment group(80 mg·mL~(-1) ox-LDL+10 μmol·L~(-1) Chr), and treatment group(80 mg·mL~(-1) ox-LDL+15 μmol·L~(-1) Chr). Lipid accumulation in each group was detected by oil red O staining. CD36 expression was analyzed by flow cytometry. Western blot was used to detect the expression of scavenger receptor class A1(SR-A1), scavenger receptor class B type Ⅰ(SR-B1), autophagy-related protein 5(Atg5), Beclin-1, autophagy adaptor protein p62(P62), the ratio of microtubule-associated protein light chain 3(LC3)Ⅱ to LC3Ⅰ(LC3Ⅱ/LC3Ⅰ), nuclear factor kappa B P65(NF-κB P65), inhibitor of κB kinase β(IKKβ), nuclear factor of κB inhibitor(IκB), high mobility group box protein 1(HMGB1), phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), and phosphorylated mammalian target of rapamycin(mTOR). Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression levels of ATP-binding cassette transporter A1(ABCA1), ATP-binding cassette transporter G1(ABCG1), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), HMGB1, inducible nitric oxide synthase(iNOS), arginase 1(Arg1), macrophage galactose-type lectin-1(Mgl-1), and NF-κB P65. Immunofluorescence analysis was performed to determine the localization of HMGB1 in RAW264.7 cells in each group. The autophagy inhibitor 3-methyladenine(3-MA) was added as a control for reverse validation, and the RAW264.7 macrophages were divided into four groups again: control group, model group(80 mg·mL~(-1) ox-LDL), treatment group(80 mg·mL~(-1) ox-LDL + 15 μmol·L~(-1) Chr), and inhibitor group(80 mg·mL~(-1) ox-LDL+15 μmol·L~(-1) Chr+3-MA). The results showed that Chr effectively reduced foam cell formation by regulating the expression levels of SR-A1, ABCA1, ABCG1, the LC3Ⅱ/LC3Ⅰ ratio, Atg5, Beclin-1, and p62, and inhibited the NF-κB/HMGB1-PI3K/Akt/mTOR signaling pathway. Moreover, the inhibitory effects of Chr on autophagy and the NF-κB/HMGB1-PI3K/Akt/mTOR pathway were reversed by the autophagy inhibitor 3-MA. In conclusion, Chr exhibits therapeutic potential for the treatment of atherosclerosis by inducing autophagy and modulating the NF-κB/HMGB1 and PI3K/Akt/mTOR pathways to inhibit the formation of macrophage infla","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6439-6449"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p><p>This study aims to investigate the mechanism of tanshinone Ⅱ_A(Tan Ⅱ_A) in protecting mice from diethylinitrosamine(DEN)/carbon tetrachloride(CCl_4)/ethanol(C_2H_5OH)-induced hepatocellular carcinoma(HCC) and HepG2 cells from hydrogen peroxide(H_2O_2)-induced oxidative damage via the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt) and nuclear factor E2-related factor 2(Nrf2)/heme oxygenase 1(HO-1) signaling pathways. Sixty male C57BL/6J mice were grouped as follows: control, model, low, medium, and high-dose(10, 20, 40 mg·kg~(-1), respectively) Tan Ⅱ_A, and colchicine(0.2 mg·kg~(-1)), with 10 mice in each group. The HCC model was established with DEN/CCl_4/C_2H_5OH for 20 weeks, and the mice were then euthanized for collection of blood and liver specimens. A protein-protein interaction(PPI) network of the targets of Tan Ⅱ_A in the prevention of HCC was constructed. HepG2 cells were treated with 150 μmol·L~(-1) H_2O_2 for the modeling of oxidative stress. The cell counting kit-8(CCK-8) was used to assess the effects of different concentrations(1, 2, 4 μmol·L~(-1)) of Tan Ⅱ_A on the relative viability of cells. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in the liver tissue, and an automatic biochemical analyzer was used to measure the levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the mice serum. Relevant assay kits were used to measure the levels of reactive oxygen species(ROS), malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and glutathione peroxidase(GSH-Px) in the liver and cells. Immunohistochemistry, immunofluorescence assay, and Western blot were employed to determine the expression of phosphorylated PI3K(p-PI3K), PI3K, phosphorylated Akt(p-Akt), Akt, Nrf2, and HO-1. Compared with the control group, the model group exhibited typical pathological manifestations of liver cancer in the liver tissue, with elevated levels of ALT and AST in the serum, risen levels of ROS and MDA in the liver, and lowered levels of SOD and GSH in the liver. Compared with the control group, the HepG2 cells treated with H_2O_2 showed significantly decreased activities of SOD and GSH-Px and increased ROS and MDA levels. In the liver tissue and HepG2 cells, the modeling up-regulated the expression of p-PI3K and p-Akt while down-regulating the expression of Nrf2 and HO-1. Compared with the model group, different doses of Tan Ⅱ_A reduced the levels of ALT and AST in the serum, elevated the levels of SOD, GSH, and GSH-Px in the liver tissue and cells, and lowered the ROS and MDA levels, which indicated significant alleviation of oxidative stress. The PPI network showed that Akt was a core target of the PI3K/Akt and Nrf2/HO-1 signaling pathways. Western blot, immunohistochemistry, and immunofluorescence assay results indicated that Tan Ⅱ_A promoted the expression of Nrf2 and HO-1 while inhibiting the phosphorylation of PI3K and Akt. In conclusion, Tan Ⅱ_A may delay the progressi
{"title":"[Tanshinone Ⅱ_A exerts anti-hepatocellular carcinoma effects by inhibiting oxidative stress via PI3K/Akt and Nrf2/HO-1 signaling pathway].","authors":"Meng Fang, Dong-Rui Huang, Jin-Wei Zhang, Wen-Jing Liao, Fang Wu, Yu-Wei Liu","doi":"10.19540/j.cnki.cjcmm.20240711.401","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240711.401","url":null,"abstract":"<p><p>This study aims to investigate the mechanism of tanshinone Ⅱ_A(Tan Ⅱ_A) in protecting mice from diethylinitrosamine(DEN)/carbon tetrachloride(CCl_4)/ethanol(C_2H_5OH)-induced hepatocellular carcinoma(HCC) and HepG2 cells from hydrogen peroxide(H_2O_2)-induced oxidative damage via the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt) and nuclear factor E2-related factor 2(Nrf2)/heme oxygenase 1(HO-1) signaling pathways. Sixty male C57BL/6J mice were grouped as follows: control, model, low, medium, and high-dose(10, 20, 40 mg·kg~(-1), respectively) Tan Ⅱ_A, and colchicine(0.2 mg·kg~(-1)), with 10 mice in each group. The HCC model was established with DEN/CCl_4/C_2H_5OH for 20 weeks, and the mice were then euthanized for collection of blood and liver specimens. A protein-protein interaction(PPI) network of the targets of Tan Ⅱ_A in the prevention of HCC was constructed. HepG2 cells were treated with 150 μmol·L~(-1) H_2O_2 for the modeling of oxidative stress. The cell counting kit-8(CCK-8) was used to assess the effects of different concentrations(1, 2, 4 μmol·L~(-1)) of Tan Ⅱ_A on the relative viability of cells. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in the liver tissue, and an automatic biochemical analyzer was used to measure the levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the mice serum. Relevant assay kits were used to measure the levels of reactive oxygen species(ROS), malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and glutathione peroxidase(GSH-Px) in the liver and cells. Immunohistochemistry, immunofluorescence assay, and Western blot were employed to determine the expression of phosphorylated PI3K(p-PI3K), PI3K, phosphorylated Akt(p-Akt), Akt, Nrf2, and HO-1. Compared with the control group, the model group exhibited typical pathological manifestations of liver cancer in the liver tissue, with elevated levels of ALT and AST in the serum, risen levels of ROS and MDA in the liver, and lowered levels of SOD and GSH in the liver. Compared with the control group, the HepG2 cells treated with H_2O_2 showed significantly decreased activities of SOD and GSH-Px and increased ROS and MDA levels. In the liver tissue and HepG2 cells, the modeling up-regulated the expression of p-PI3K and p-Akt while down-regulating the expression of Nrf2 and HO-1. Compared with the model group, different doses of Tan Ⅱ_A reduced the levels of ALT and AST in the serum, elevated the levels of SOD, GSH, and GSH-Px in the liver tissue and cells, and lowered the ROS and MDA levels, which indicated significant alleviation of oxidative stress. The PPI network showed that Akt was a core target of the PI3K/Akt and Nrf2/HO-1 signaling pathways. Western blot, immunohistochemistry, and immunofluorescence assay results indicated that Tan Ⅱ_A promoted the expression of Nrf2 and HO-1 while inhibiting the phosphorylation of PI3K and Akt. In conclusion, Tan Ⅱ_A may delay the progressi","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 24","pages":"6724-6734"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In this study, the chemical components of Panacis Japonici Rhizoma extract and absorbed components in rats were identified by ultra-high performance liquid chromatography-quadrupole exactive orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS). The separation was performed by gradient elution on Waters UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of water and acetonitrile containing 0.1% formic acid. High resolution multistage mass spectrometry data were collected by electrospray ionization in positive and negative ion modes. The chemical components of Panacis Japonici Rhizoma extract were identified by comparing with the retention time, high resolution precise molecular weight, and secondary fragment ions of reference substances and related literature. After intragastric administration of Panacis Japonici Rhizoma extract, blood was collected from the abdominal aorta of rats for separation of the serum, and the absorbed components were scanned and identified. The results showed that 43 chemical components were detected in the Panacis Japonici Rhizoma extract, including 22 saponins, 9 amino acids, 5 polysaccharides, 2 volatile oils, and 5 nucleosides. In the serum, 18 components were detected, including 10 prototype components, 6 metabolites, and 2 unknown components. This study analyzed the chemical components and absorbed components of Panacis Japonici Rhizoma extract, providing clues for clarifying the pharmacological basis of Panacis Japonici Rhizoma.
{"title":"[Serum pharmacochemistry of Panacis Japonici Rhizoma extract based on UPLC-Q-Exactive Orbitrap-MS].","authors":"Yuan Wen, Chang Liu, Qi Yuan, Rui Wang, Xue-Qing Li, Ding Yuan, Cheng-Fu Yuan, Yu-Min He","doi":"10.19540/j.cnki.cjcmm.20240910.203","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240910.203","url":null,"abstract":"<p><p>In this study, the chemical components of Panacis Japonici Rhizoma extract and absorbed components in rats were identified by ultra-high performance liquid chromatography-quadrupole exactive orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS). The separation was performed by gradient elution on Waters UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of water and acetonitrile containing 0.1% formic acid. High resolution multistage mass spectrometry data were collected by electrospray ionization in positive and negative ion modes. The chemical components of Panacis Japonici Rhizoma extract were identified by comparing with the retention time, high resolution precise molecular weight, and secondary fragment ions of reference substances and related literature. After intragastric administration of Panacis Japonici Rhizoma extract, blood was collected from the abdominal aorta of rats for separation of the serum, and the absorbed components were scanned and identified. The results showed that 43 chemical components were detected in the Panacis Japonici Rhizoma extract, including 22 saponins, 9 amino acids, 5 polysaccharides, 2 volatile oils, and 5 nucleosides. In the serum, 18 components were detected, including 10 prototype components, 6 metabolites, and 2 unknown components. This study analyzed the chemical components and absorbed components of Panacis Japonici Rhizoma extract, providing clues for clarifying the pharmacological basis of Panacis Japonici Rhizoma.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6490-6499"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The chemical constituents from the stems and leaves of Mycetia hainanensis were isolated by silica gel, ODS gel, and Sephadex LH-20 gel column chromatography and preparative HPLC. The chemical structures of all the isolated compounds were identified on the basis of their physicochemical properties, spectroscopic data, as well as the comparison of their physicochemical and spectroscopic data with the reported data in literature. Twelve compounds were isolated from the 85% ethanol extract of the stems and leaves of M. hainanensis, and they were identified as mycehainanic acid(1), 11-methoxyviburtinal(2), isovaltrate acetoxyhydrin(3), jatamanvaltrate K(4), jatamanin C(5), sarmentol F(6), 4,5-dihydroblumenol A(7), petasitolone(8), xylaguaianol D(9), aristolan-9-en-1-one(10), 3α-hydroxycostic acid(11), and ilicic acid(12). Among them, compound 1 was a new nor-sesquiterpene, and compounds 2-12 were isolated from Mycetia for the first time. In addition, the MTS method was employed to assess the anti-rheumatoid arthritis activities of compounds 1-12 based on their anti-proliferative effects on synoviocytes in vitro. The results showed that compounds 1 and 6-12 exhibited notable anti-rheumatoid arthritis activities, showcasing inhibitory effects on the proliferation of MH7A synovial fibroblast cells with the IC_(50) ranging from(5.27±0.07) to(172.68±0.32)μmol·L~(-1), which were comparable to that of the positive control methotrexate [IC_(50) of(132.39±0.21)μmol·L~(-1)].
采用硅胶柱层析、ODS凝胶柱层析、Sephadex LH-20凝胶柱层析和制备高效液相色谱法对海南支原体茎叶的化学成分进行分离。根据化合物的理化性质、光谱学数据以及与文献报道的理化、光谱学数据的比较,确定了化合物的化学结构。从85个化合物中分离得到12个化合物% ethanol extract of the stems and leaves of M. hainanensis, and they were identified as mycehainanic acid(1), 11-methoxyviburtinal(2), isovaltrate acetoxyhydrin(3), jatamanvaltrate K(4), jatamanin C(5), sarmentol F(6), 4,5-dihydroblumenol A(7), petasitolone(8), xylaguaianol D(9), aristolan-9-en-1-one(10), 3α-hydroxycostic acid(11), and ilicic acid(12). Among them, compound 1 was a new nor-sesquiterpene, and compounds 2-12 were isolated from Mycetia for the first time. In addition, the MTS method was employed to assess the anti-rheumatoid arthritis activities of compounds 1-12 based on their anti-proliferative effects on synoviocytes in vitro. The results showed that compounds 1 and 6-12 exhibited notable anti-rheumatoid arthritis activities, showcasing inhibitory effects on the proliferation of MH7A synovial fibroblast cells with the IC_(50) ranging from(5.27±0.07) to(172.68±0.32)μmol·L~(-1), which were comparable to that of the positive control methotrexate [IC_(50) of(132.39±0.21)μmol·L~(-1)].
{"title":"[Chemical constituents from stems and leaves of Mycetia hainanensis and their anti-rheumatoid arthritis activities].","authors":"Xiao-Ya Fu, Jia-Yi Hu, Jing-Su Yu, Yan-Zhi Liu, Ke-Jin Jiang, Bing-Yan Sun, Yan-Ping Liu, Yan-Hui Fu","doi":"10.19540/j.cnki.cjcmm.20240926.201","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240926.201","url":null,"abstract":"<p><p>The chemical constituents from the stems and leaves of Mycetia hainanensis were isolated by silica gel, ODS gel, and Sephadex LH-20 gel column chromatography and preparative HPLC. The chemical structures of all the isolated compounds were identified on the basis of their physicochemical properties, spectroscopic data, as well as the comparison of their physicochemical and spectroscopic data with the reported data in literature. Twelve compounds were isolated from the 85% ethanol extract of the stems and leaves of M. hainanensis, and they were identified as mycehainanic acid(1), 11-methoxyviburtinal(2), isovaltrate acetoxyhydrin(3), jatamanvaltrate K(4), jatamanin C(5), sarmentol F(6), 4,5-dihydroblumenol A(7), petasitolone(8), xylaguaianol D(9), aristolan-9-en-1-one(10), 3α-hydroxycostic acid(11), and ilicic acid(12). Among them, compound 1 was a new nor-sesquiterpene, and compounds 2-12 were isolated from Mycetia for the first time. In addition, the MTS method was employed to assess the anti-rheumatoid arthritis activities of compounds 1-12 based on their anti-proliferative effects on synoviocytes in vitro. The results showed that compounds 1 and 6-12 exhibited notable anti-rheumatoid arthritis activities, showcasing inhibitory effects on the proliferation of MH7A synovial fibroblast cells with the IC_(50) ranging from(5.27±0.07) to(172.68±0.32)μmol·L~(-1), which were comparable to that of the positive control methotrexate [IC_(50) of(132.39±0.21)μmol·L~(-1)].</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 24","pages":"6692-6698"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study primarily investigated the mechanism of Astragalus polysaccharides(APS), a Chinese medicinal material, in regulating the Nrf2/SLC7A11/GPX4 signaling pathway to induce ferroptosis in ovarian cancer cells(Caov-3 and SKOV3 cells). Caov-3 and SKOV3 cells were divided into control(Vehicle) group, APS group, glutathione peroxidase 4 inhibitor(RSL3) group, and APS+RSL3 group. After 48 h of intervention, the activity and morphology of the cells in each group were observed. The cell counting kit-8(CCK-8) method was used to determine the half-maximal inhibitory concentration(IC_(50)), while colony formation and EdU assays were conducted to assess cell proliferation. Biochemical reagents were used to detect lipid reactive oxygen species(L-ROS), malondialdehyde(MDA), divalent iron ions(Fe~(2+)), and glutathione(GSH) in Caov-3 cells. Transmission electron microscopy was employed to observe the morphological changes of mitochondria in Caov-3 cells. Bioinformatics analysis were used to screen potential target genes of APS in ovarian cancer cells. Western blot and RT-PCR were applied to measure the protein and mRNA expression of Nrf2, SLC7A11, and GPX4. The results revealed that APS effectively inhibited the activity and proliferation of ovarian cancer cells, significantly increased the expression levels of L-ROS, MDA, and Fe~(2+)(P<0.001), and significantly reduced the expression level of GSH(P<0.001). Under electron microscopy, the mitochondria of Caov-3 cells appeared significantly smaller, with a marked increase in the density of the bilayer membrane, disappearance of mitochondrial cristae, and rupture of the outer mitochondrial membrane. These effects were more pronounced when APS was combined with RSL3. Bioinformatics screening identified Nrf2, SLC7A11, and GPX4 as potential target genes for APS in ovarian cancer cells. APS was shown to reduce the protein and mRNA expression of Nrf2, SLC7A11, and GPX4(P<0.01), with the APS+RSL3 showing even more significant effects(P<0.001). In conclusion, APS can induce ferroptosis in ovarian cancer cells, and its mechanism may be related to the regulation of the Nrf2/SLC7A11/GPX4 signaling pathway, providing an experimental basis for the use of APS injections in the treatment of ovarian cancer.
{"title":"[Astragalus polysaccharides induces ferroptosis in ovarian adenocarcinoma cells through Nrf2/SLC7A11/GPX4 signaling pathway].","authors":"Yong-Gen Zhang, Xiao-Fei Yan, Feng Liu, Wen-Zhe Hao, Yue Cai, Ying Liu, Lan-Lin Liu, Xue-Jun Li","doi":"10.19540/j.cnki.cjcmm.20240919.501","DOIUrl":"https://doi.org/10.19540/j.cnki.cjcmm.20240919.501","url":null,"abstract":"<p><p>This study primarily investigated the mechanism of Astragalus polysaccharides(APS), a Chinese medicinal material, in regulating the Nrf2/SLC7A11/GPX4 signaling pathway to induce ferroptosis in ovarian cancer cells(Caov-3 and SKOV3 cells). Caov-3 and SKOV3 cells were divided into control(Vehicle) group, APS group, glutathione peroxidase 4 inhibitor(RSL3) group, and APS+RSL3 group. After 48 h of intervention, the activity and morphology of the cells in each group were observed. The cell counting kit-8(CCK-8) method was used to determine the half-maximal inhibitory concentration(IC_(50)), while colony formation and EdU assays were conducted to assess cell proliferation. Biochemical reagents were used to detect lipid reactive oxygen species(L-ROS), malondialdehyde(MDA), divalent iron ions(Fe~(2+)), and glutathione(GSH) in Caov-3 cells. Transmission electron microscopy was employed to observe the morphological changes of mitochondria in Caov-3 cells. Bioinformatics analysis were used to screen potential target genes of APS in ovarian cancer cells. Western blot and RT-PCR were applied to measure the protein and mRNA expression of Nrf2, SLC7A11, and GPX4. The results revealed that APS effectively inhibited the activity and proliferation of ovarian cancer cells, significantly increased the expression levels of L-ROS, MDA, and Fe~(2+)(P<0.001), and significantly reduced the expression level of GSH(P<0.001). Under electron microscopy, the mitochondria of Caov-3 cells appeared significantly smaller, with a marked increase in the density of the bilayer membrane, disappearance of mitochondrial cristae, and rupture of the outer mitochondrial membrane. These effects were more pronounced when APS was combined with RSL3. Bioinformatics screening identified Nrf2, SLC7A11, and GPX4 as potential target genes for APS in ovarian cancer cells. APS was shown to reduce the protein and mRNA expression of Nrf2, SLC7A11, and GPX4(P<0.01), with the APS+RSL3 showing even more significant effects(P<0.001). In conclusion, APS can induce ferroptosis in ovarian cancer cells, and its mechanism may be related to the regulation of the Nrf2/SLC7A11/GPX4 signaling pathway, providing an experimental basis for the use of APS injections in the treatment of ovarian cancer.</p>","PeriodicalId":52437,"journal":{"name":"Zhongguo Zhongyao Zazhi","volume":"49 23","pages":"6459-6467"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}