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Intervention effect of Cigu Xiaozhi prescription on ceramide lipoapoptosis in non-alcoholic fatty liver disease. 慈姑小柴胡汤对非酒精性脂肪肝神经酰胺脂质凋亡的干预作用
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231215.002
Yang Shaojun, M A Yanhua, Bai Zhouxia, Y U Ye, Fang Buwu, Zhang Li, Wang Li

Objective: To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription (, CGXZ) in the treatment of the non-alcoholic fatty liver disease (NAFLD) by detoxification and phlegm-reducing, the effect of CGXZ prescription on ceramide-mediated lipid apoptosis in Hep G2 cells with NAFLD.

Methods: The experiment was randomly divided into 6 groups: normal control group, model group, CGXZ prescription medicated serum high, medium, and low dose groups, and pioglitazone positive control group. Using 500 μmol/L free fatty acid (FFA) mixture to induce Hep G2 cells to establish NAFLD cell model, respectively, with 2%, 4%, and 6% concentration of CGXZ prescription medicated serum intervention for 24 h. The changes in organelles and lipid droplet accumulation were observed under the electron microscope. Furthermore, TdT-mediated dUTP Nick-End Labeling method was used to assay hepatocyte apoptosis; Biochemical determination of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase, triglycerides, and FFA levels in Hep G2 cells; the content of ceramide was determined by high-performance thin-layer chromatography. Finally, Western Blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to determine the protein and gene expression levels, such as inducible nitric oxide synthase (iNOS), nuclear factor κB (NF-κB), B cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax).

Results: Under the electron microscope, the cells in the model group showed moderate-to-severe steatosis, and apoptotic bodies could be seen. The model group had greater improvements in the apoptosis rate (P < 0.01), and the levels of ceramide C2 and FFA in the cytoplasm (P < 0.01) than the normal control group. The protein expressions of NF-κB, iNOS, and Bax were significantly up-regulated (P < 0.05), while the Bcl-2 had no significant change (P > 0.05). Compared with the model group, the levels of ceramide C2 and FFA (P < 0.01), the protein expressions of NF-κB, iNOS, and Bax (P < 0.05) in the CGXZ prescription treatment group and pioglitazone positive control group were significantly decreased; Only the Bcl-2 protein was significantly up-regulated in the high-dose Chinese medicine group (P < 0.05). The down-regulation of Bax mRNA expression in each Chinese medicine treatment group was significantly better than in the pioglitazone positive control group (P < 0.01).

Conclusions: The CGXZ prescription, formulated with the method of detoxification and phlegm, can inhibit lipoapoptosis in the NAFLD cell model by down-regulating the levels of ceramide C2 and FFA, which may be achieved by regulating ceramide/iNOS/NF-κB signaling pathway.

研究目的探讨中药慈姑小柴胡汤(CGXZ)解毒化痰治疗非酒精性脂肪肝的机理,以及CGXZ方剂对非酒精性脂肪肝Hep G2细胞神经酰胺介导的脂质凋亡的影响:实验随机分为6组:正常对照组,模型组,CGXZ处方药物血清高、中、低剂量组,吡格列酮阳性对照组。用500 μmol/L游离脂肪酸(FFA)混合液诱导Hep G2细胞建立非酒精性脂肪肝细胞模型,分别用2%、4%和6%浓度的CGXZ处方药物血清干预24 h,电镜下观察细胞器的变化和脂滴的聚集。此外,还采用TdT介导的dUTP镍末端标记法测定肝细胞凋亡;生化法测定Hep G2细胞谷丙转氨酶、谷草转氨酶、甘油三酯和FFA的水平;采用高效薄层色谱法测定神经酰胺的含量。最后,采用 Western Blot 和实时定量聚合酶链反应(qRT-PCR)检测诱导型一氧化氮合酶(iNOS)、核因子κB(NF-κB)、B 细胞淋巴瘤 2(Bcl-2)和 Bcl-2 相关 X(Bax)等蛋白质和基因的表达水平:在电子显微镜下,模型组细胞出现中度至重度脂肪变性,并可见凋亡体。与正常对照组相比,模型组细胞凋亡率(P 0.01)、细胞质中神经酰胺 C2 和 FFA 的水平(P 0.01)均有较大改善。NF-κB、iNOS和Bax的蛋白表达明显上调(P 0.05),而Bcl-2无明显变化(P > 0.05)。与模型组相比,CGXZ处方治疗组和吡格列酮阳性对照组的神经酰胺C2和FFA水平(P 0.01)、NF-κB、iNOS和Bax蛋白表达量(P 0.05)均明显下降;只有大剂量中药组的Bcl-2蛋白明显上调(P 0.05)。各中药治疗组Bax mRNA表达下调情况明显优于吡格列酮阳性对照组(P 0.01):以解毒化痰法配伍的CGXZ方剂可通过下调神经酰胺C2和FFA的水平抑制非酒精性脂肪肝细胞模型的脂肪凋亡,而这可能是通过调节神经酰胺/iNOS/NF-κB信号通路实现的。
{"title":"Intervention effect of Cigu Xiaozhi prescription on ceramide lipoapoptosis in non-alcoholic fatty liver disease.","authors":"Yang Shaojun, M A Yanhua, Bai Zhouxia, Y U Ye, Fang Buwu, Zhang Li, Wang Li","doi":"10.19852/j.cnki.jtcm.20231215.002","DOIUrl":"10.19852/j.cnki.jtcm.20231215.002","url":null,"abstract":"<p><strong>Objective: </strong>To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription (, CGXZ) in the treatment of the non-alcoholic fatty liver disease (NAFLD) by detoxification and phlegm-reducing, the effect of CGXZ prescription on ceramide-mediated lipid apoptosis in Hep G2 cells with NAFLD.</p><p><strong>Methods: </strong>The experiment was randomly divided into 6 groups: normal control group, model group, CGXZ prescription medicated serum high, medium, and low dose groups, and pioglitazone positive control group. Using 500 μmol/L free fatty acid (FFA) mixture to induce Hep G2 cells to establish NAFLD cell model, respectively, with 2%, 4%, and 6% concentration of CGXZ prescription medicated serum intervention for 24 h. The changes in organelles and lipid droplet accumulation were observed under the electron microscope. Furthermore, TdT-mediated dUTP Nick-End Labeling method was used to assay hepatocyte apoptosis; Biochemical determination of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase, triglycerides, and FFA levels in Hep G2 cells; the content of ceramide was determined by high-performance thin-layer chromatography. Finally, Western Blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to determine the protein and gene expression levels, such as inducible nitric oxide synthase (iNOS), nuclear factor κB (NF-κB), B cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax).</p><p><strong>Results: </strong>Under the electron microscope, the cells in the model group showed moderate-to-severe steatosis, and apoptotic bodies could be seen. The model group had greater improvements in the apoptosis rate (<i>P <</i> 0.01), and the levels of ceramide C2 and FFA in the cytoplasm (<i>P <</i> 0.01) than the normal control group. The protein expressions of NF-κB, iNOS, and Bax were significantly up-regulated (<i>P <</i> 0.05), while the Bcl-2 had no significant change (<i>P ></i> 0.05). Compared with the model group, the levels of ceramide C2 and FFA (<i>P <</i> 0.01), the protein expressions of NF-κB, iNOS, and Bax (<i>P <</i> 0.05) in the CGXZ prescription treatment group and pioglitazone positive control group were significantly decreased; Only the Bcl-2 protein was significantly up-regulated in the high-dose Chinese medicine group (<i>P <</i> 0.05). The down-regulation of Bax mRNA expression in each Chinese medicine treatment group was significantly better than in the pioglitazone positive control group (<i>P <</i> 0.01).</p><p><strong>Conclusions: </strong>The CGXZ prescription, formulated with the method of detoxification and phlegm, can inhibit lipoapoptosis in the NAFLD cell model by down-regulating the levels of ceramide C2 and FFA, which may be achieved by regulating ceramide/iNOS/NF-κB signaling pathway.</p>","PeriodicalId":94119,"journal":{"name":"Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10774722/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139428189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Qingchang suppositry induced remission in patients with mild-to-moderate ulcerative proctitis: a multicenter, prospective, randomized, parallel-controlled clinical trial. 清畅栓诱导轻中度溃疡性直肠炎患者病情缓解:一项多中心、前瞻性、随机、平行对照临床试验。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231121.004
Dai Xiaoling, Zhang Anming, Lin Hui, Shi Bei, Ren Yi, Wen Hongzhu, Fei Xiaoyan, Lin Jiang

Objective: To evaluate the efficacy and safety of Qingchang suppository (, QCS), a preparation of Chinese herbal medicine, in the induction of remission in patients with mild-to-moderate ulcerative proctitis (UP).

Methods: We performed a multicenter, prospective, randomized, parallel-controlled trial to evaluate the efficacy of QCS induction therapy in 140 adult patients with mild-to-moderate UP and TCM syndrome of dampness-heat in large intestine. The patients were randomized to receive QCS (study group) or Salicylazosulfapyridine (SASP) suppository (control group) one piece each time, twice a day, per anum for 12 weeks. Mayo score and main symptoms score were evaluated at weeks 0, 2, 4, 8 and 12, rectosigmoidscopy was taken at weeks 0, 4, 8 and 12, Geboes score, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and safety indexes were assessed at weeks 0 and 12. The primary efficacy endpoint is clinical remission rate, the secondary efficacy endpoints are clinical response rate, mucosa healing rate, Geboes score, the remission rates of the main symptoms, the median day to the remission of the symptom, etc. RESULTS: There were no statistical difference in the clinical remission rates, the clinical response rates, the mucosa healing rates, Geboes score, ESR and CRP between the two groups. The remission rates of tenesmus and anal burning sensation of the study group were significantly higher than those of the control group (76.5% vs 25.0%, P = 0.009; 74.51% vs 29.63%, P = 0.003). The median day to the remission of purulent bloody stool of the study group was significantly less than that of control group [11 (1, 64) vs 19 (2, 67), P = 0.007]. The patients receiving QCS had a significantly higher mucosa healing rate at week 4 than the patients receiving SASP suppository (71.42% vs 52.85%, P = 0.023). No adverse event occurred in the study group while the adverse events incidence of the control group was 5.7% (P = 0.049).

Conclusions: QCS could induce the remission of UP as effectively and safely as SASP suppository, and was superior to SASP suppository in relieving the symptoms of tenesmus, anal burning sensation and purulent bloody stool and the time to reach mucosa healing.

目的评价清畅栓剂(一种中药制剂)诱导轻中度溃疡性直肠炎(UP)患者病情缓解的疗效和安全性:我们进行了一项多中心、前瞻性、随机、平行对照试验,以评估QCS诱导疗法对140例轻中度溃疡性直肠炎和中医大肠湿热证成人患者的疗效。患者随机接受 QCS(研究组)或柳氮磺吡啶栓剂(对照组)治疗,每次一粒,每天两次,经肛门使用,为期 12 周。在第 0、2、4、8 和 12 周评估梅奥评分和主要症状评分,在第 0、4、8 和 12 周进行直肠乙状结肠镜检查,在第 0 和 12 周评估 Geboes 评分、红细胞沉降率(ESR)、C 反应蛋白(CRP)和安全性指标。主要疗效终点为临床缓解率,次要疗效终点为临床反应率、黏膜愈合率、Geboes评分、主要症状缓解率、症状缓解中位天数等。结果:两组患者的临床缓解率、临床反应率、黏膜愈合率、Geboes 评分、血沉和 CRP 均无统计学差异。研究组痛经和肛门烧灼感的缓解率明显高于对照组(76.5% vs 25.0%,P = 0.009;74.51% vs 29.63%,P = 0.003)。研究组患者脓血便缓解的中位天数明显少于对照组 [11 (1, 64) vs 19 (2, 67), P = 0.007]。接受 QCS 治疗的患者在第 4 周的粘膜愈合率明显高于接受 SASP 栓剂治疗的患者(71.42% vs 52.85%,P = 0.023)。研究组无不良反应发生,而对照组的不良反应发生率为5.7%(P = 0.049):结论:QCS与SASP栓剂一样能有效、安全地诱导UP缓解,在缓解胀痛、肛门烧灼感和脓血便症状以及黏膜愈合时间方面优于SASP栓剂。
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引用次数: 0
Taohong Siwu decoction ameliorates atherosclerosis in rats possibly through toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-κB signal pathway. 桃红四物汤改善大鼠动脉粥样硬化,可能是通过收费样受体 4/髓系分化初级反应蛋白 88/核因子-κB 信号通路。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231215.003
Chang Fengjin, Zhou Peng, L I Guoying, Zhang Weizhi, Zhang Yanyan, Peng Daiyin, Chen Guangliang

Objective: To investigate the effect of Taohong Siwu decoction (, TSD) on atherosclerosis in rats as well as investigate the underlying mechanism based on molecular docking.

Methods: Sixty healthy male Sprague-Dawley rats were randomly divided into 6 groups with 10 rats in each group: control group, model group, atorvastatin group (AT, 2.0 mg/kg), and TSD groups (20, 10, 5 g/kg) after 7 d of acclimation. The model of atherosclerosis was successfully established except the control group by high fat diet (HFD) and vitamin D2. Biochemical analyzers were used to detect the levels of triglyceride (TG), total cholestero (TC), low density lipoprotein-cholesterol (LDL-C) and high density lipid-cholesterol (HDL-C) in blood lipid. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were determined by enzyme-linked immunosorbent assay. Sudan IV staining and Hematoxylin and eosin staining (HE staining) were performed to observe the pathological changes in aortic tissue. Molecular docking technology was used to predict the best matching between the main components of TSD and the target proteins. The expression of target proteins was further detected by quantitative real time polymerase chain reaction (qRT-PCR) and Western blot analysis.

Results: The results showed that TSD restricted atherosclerosis development and decreased the inflammatory cytokines in plasma. Molecular docking results predicted that the main components of TSD showed a strong binding ability with toll-like receptor (TLR4), myeloid differentiation primary response protein 88 (MyD88), and nuclear factor kappa-B (NF-κB). The results of qRT-PCR and Western blot analysis showed that the mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 in the aorta were reduced in atorvastatin group and TSD group.

Conclusions: TSD can ameliorate atherosclerosis in rats, and the underlying mechanism is supposed be related to the suppression of inflammatory response by regulating TLR4/MyD88/NF-κB signal pathway.

目的研究桃红四物汤(TSD)对大鼠动脉粥样硬化的影响,并基于分子对接研究其作用机制:将60只健康雄性Sprague-Dawley大鼠随机分为6组,每组10只,分别为对照组、模型组、阿托伐他汀组(AT,2.0 mg/kg)和TSD组(20、10、5 g/kg)。除对照组外,通过高脂饮食(HFD)和维生素D2成功建立了动脉粥样硬化模型。生化分析仪检测血脂中甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的水平。肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的水平通过酶联免疫吸附试验测定。通过苏丹Ⅳ染色和苏木精及伊红染色观察主动脉组织的病理变化。利用分子对接技术预测 TSD 主要成分与目标蛋白的最佳匹配度。通过实时定量聚合酶链反应(qRT-PCR)和Western印迹分析进一步检测目标蛋白的表达:结果表明,TSD 限制了动脉粥样硬化的发展,并降低了血浆中的炎性细胞因子。分子对接结果表明,TSD的主要成分与类收费受体(TLR4)、髓样分化初级反应蛋白88(MyD88)和核因子卡巴-B(NF-κB)有很强的结合能力。qRT-PCR和Western印迹分析结果显示,阿托伐他汀组和TSD组主动脉中TLR4、MyD88和NF-κB p65的mRNA和蛋白表达量均有所下降:结论:TSD可改善大鼠动脉粥样硬化,其机制可能与通过调节TLR4/MyD88/NF-κB信号通路抑制炎症反应有关。
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引用次数: 0
Effect of nourishing and purging fire Chinese herbal mixture on delaying light-induced premature puberty in rats. 滋阴清火中药组合对延缓光照诱导的大鼠性早熟的影响
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20230814.001
Sun Yanyan, Xue Yuanyuan, Sun Wen, Wang Yonghong, Y U Jian

Objective: To elucidate the mechanism of the nourishing Yin and purging fire Chinese herbal mixture (NYPF) in delaying light-induced premature puberty in rats.

Methods: Twenty-one days old female Sprague-Dawley rats were randomly assigned to normal group (N), long light exposure group (L), NYPF and normal saline group (NS). Rats in the L, NYPF and NS groups were exposed to 16 h: 350 lux light/8 h: dark, while rats in the N group were exposed to 12 h: 50 lux light/12 h: dark. NYPF and normal saline was administered to the rats in the NYPF group or NS group, respectively, from day 21. Five rats in every group were sacrificed at 9 p.m. on day 28 (P28), on the day when rat's vulva opened in the L group (L-VO), on the day when the first estrous interphase occurred in rats of L group (L-E1), and on the day when the second estrous interphase occurred in rats of L group (L-E2), respectively.

Resulits: On day 34, all rats in the L group, 80% of rats in the NS group, 40% of rats in the N group, and 20% of rats in the NYPF group showed complete opening of the vulva. At P28, mRNA level of hypothalamic kisspeptin (Kiss-1) in the L group was significantly higher than that in the N group (P < 0.05). The rats in the L and NS groups had significantly lower hypothalamic arginine-phenylalanine-amide (RFamide)-related peptide 3 (RFRP-3) mRNA levels than those in the N group (P < 0.05), whereas RFRP-3 mRNA level was significantly higher in the NYPF group than that in the L group (P < 0.05). At L-VO, the ovarian index of the L and NS groups was significantly higher than that of the N group (P < 0.05) and estradiol (E2) level of the NYPF group was significantly lower than that of the N and NS groups (P < 0.05); hypothalamic Kiss-1 mRNA level in the L and NS groups was significantly higher than that in the N and NYPF groups (P < 0.05), whereas hypothalamic RFRP-3 mRNA level in the L, NYPF, and NS groups was significantly lower than that in the N group (P < 0.05). At L-E1, E2 level of the L and NS groups was significantly higher than that of the N group (P < 0.01), whereas it was significantly lower in the NYPF group than that of the N, L, and NS groups (P < 0.01), and serum luteinizing hormone level of the L and NS groups was significantly higher than that of the N group (P < 0.05); levels of serum melatonin and ovarian melatonin receptor 1 (MT-1) mRNA in the L, NYPF, and NS groups were significantly lower than those in the N group (P < 0.05). At L-E2, the uterine organ index of the NYPF group was significantly lower than that of the L group (P < 0.05); and ovarian MT-1 mRNA level of the L and NS groups was significantly lower than that in the N group (P < 0.05).

Conclusions: NYPF can delay puberty onset in rats exposed to strong light for a prolonged duration, and regulation of the gen

目的阐明滋阴清火中药复方制剂(NYPF)延缓光照诱导的大鼠性早熟的机制:将21天大的雌性Sprague-Dawley大鼠随机分为正常组(N)、长光照组(L)、NYPF组和生理盐水组(NS)。L组、NYPF组和NS组的大鼠接受16 h:光照时间为 16 小时:350 勒克斯/8 小时:暗,而 N 组大鼠的光照时间为 12 小时:50 勒克斯/12 小时:暗。从第 21 天起,分别给 NYPF 组或 NS 组大鼠注射 NYPF 和生理盐水。第28天(P28),L组大鼠外阴张开之日(L-VO),L组大鼠第一次发情间期之日(L-E1),L组大鼠第二次发情间期之日(L-E2),每组各5只大鼠,于晚上9点处死:第34天,L组所有大鼠、NS组80%大鼠、N组40%大鼠和NYPF组20%大鼠的外阴完全张开。P28时,L组下丘脑吻肽(Kiss-1)的mRNA水平显著高于N组(P 0.05)。L组和NS组大鼠下丘脑精氨酸-苯丙氨酸酰胺(RFamide)相关肽3(RFRP-3)mRNA水平明显低于N组(P 0.05),而NYPF组RFRP-3 mRNA水平明显高于L组(P 0.05)。在L-VO时,L组和NS组的卵巢指数明显高于N组(P 0.05),NYPF组的雌二醇(E2)水平明显低于N组和NS组(P 0.05);L组和NS组下丘脑Kiss-1 mRNA水平明显高于N组和NYPF组(P 0.05),而L组、NYPF组和NS组下丘脑RFRP-3 mRNA水平明显低于N组(P 0.05)。在L-E1时,L组和NS组的E2水平明显高于N组(P 0.01),而NYPF组则明显低于N组、L组和NS组(P 0.01),L组和NS组血清促黄体生成素水平明显高于N组(P 0.05);L组、NYPF组和NS组血清褪黑素和卵巢褪黑素受体1(MT-1)mRNA水平明显低于N组(P 0.05)。在L-E2期,NYPF组的子宫器官指数明显低于L组(P 0.05);L组和NS组的卵巢MT-1 mRNA水平明显低于N组(P 0.05):结论:NYPF能延缓长期暴露于强光下的大鼠青春期的到来,其机制之一是调节下丘脑中Kiss-1和RFRP-3基因的表达。
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引用次数: 0
Efficacy of Jiangzhi Xiaoban tablet on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet. 江孜消斑片对高脂饮食诱导的动脉粥样硬化小鼠的toll样受体4/核因子-kappa B/nod样受体蛋白3信号通路的疗效。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231121.002
Liu Huihui, Feng Jun, Liu Jianhe, Cheng Choufu, H U Guoheng

Objective: To study the effect of Jiangzhi Xiaoban tablet (, JZXB) on toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB)/Nod-like receptor protein 3 (NLRP3) signaling pathway expression in atherosclerosis (AS) mice by establishing a mouse model of AS, and to explore its mechanism of prevention and treatment of AS.

Methods: Sixty-four male C57BL/6J mice were randomly divided into two groups, 12 in the normal control group and 52 in the model group (MOD). Seven weeks later, two mice in each of the above two groups were randomly sacrificed, and the whole aortic tissue of the mice was taken out for hematoxylin-eosin staining. After successful modeling, 50 mice in the modeling group were randomly divided into 5 groups: MOD, atorvastatin group (ATO), low-dose group of JZXB (JZXB-L), middle-dose group of JZXB (JZXB-M), and high-dose group of JZXB (JZXB-H), 10 mice in each group. The mice in each group were killed after 6 weeks of preventive administration. HE staining was used to observe the pathological changes of aorta in AS mice. The levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were detected by automatic biochemical analyzer. The levels of inflammatory factor interleukin-1β (IL-1β) were detected by enzyme linked immunosorbent assay. The expression of TLR4, NF-κB and NLRP3 proteins in aortic tissue was detected by immunohistochemistry.

Results: Compared with the MOD, the levels of serum TC, TG and LDL-C in the JZXB-H and ATO were significantly decreased, while the level of HDL-C was significantly increased. The levels of serum TG, LDL-C in the JZXB-M were significantly decreased, and the level of HDL-C was significantly increased. Compared with the MOD, the levels of IL-1β were significantly decreased, aortic lesions were significantly improved, and the expression of TLR4, NF-κB, and NLRP3 proteins in the aortic tissue was significantly decreased in the JZXB-H, JZXB-M, and ATO.

Conclusion: JZXB has inhibitory effect on atherosclerosis in mice, and its mechanism may be through regulating the TLR4/NF-κB/NLRP3 signaling pathway and reducing the inflammatory response, so as to play a role in inhibiting atherosclerosis.

研究目的通过建立动脉粥样硬化(AS)小鼠模型,研究江浙夏枯草片(JZXB)对动脉粥样硬化(AS)小鼠体内toll样受体4(TLR4)/核因子-kappa B(NF-κB)/Nod样受体蛋白3(NLRP3)信号通路表达的影响,并探讨其防治AS的机制:将64只雄性C57BL/6J小鼠随机分为两组,正常对照组12只,模型组(MOD)52只。七周后,随机处死上述两组各两只小鼠,取出小鼠的整个主动脉组织进行苏木精-伊红染色。建模成功后,将建模组的 50 只小鼠随机分为 5 组:MOD组、阿托伐他汀组(ATO)、JZXB低剂量组(JZXB-L)、JZXB中剂量组(JZXB-M)、JZXB高剂量组(JZXB-H),每组10只。各组小鼠在预防性给药 6 周后处死。HE 染色观察 AS 小鼠主动脉的病理变化。用自动生化分析仪检测血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的水平。酶联免疫吸附试验检测了炎症因子白细胞介素-1β(IL-1β)的水平。免疫组化法检测了主动脉组织中 TLR4、NF-κB 和 NLRP3 蛋白的表达:结果:与 MOD 相比,JZXB-H 和 ATO 的血清 TC、TG 和 LDL-C 水平显著降低,而 HDL-C 水平显著升高。JZXB-M的血清TG、LDL-C水平明显下降,HDL-C水平明显上升。与MOD相比,JZXB-H、JZXB-M和ATO的IL-1β水平明显降低,主动脉病变明显改善,主动脉组织中TLR4、NF-κB和NLRP3蛋白的表达明显减少:结论:JZXB对小鼠动脉粥样硬化有抑制作用,其机制可能是通过调节TLR4/NF-κB/NLRP3信号通路,减轻炎症反应,从而起到抑制动脉粥样硬化的作用。
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引用次数: 0
Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis. 奈非林通过调节 miR-17-5p/ 核因子 E2 相关因子 2 轴抑制糖尿病肾病的进展。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231204.004
Huang Hongmei, Yang Maojun, L I Ting, Wang Dandan, L I Ying, Tang Xiaochi, Yuan Lu, G U Shi, X U Yong

Objective: To investigate the effect of Neferine (Nef) on diabetic nephropathy (DN) and to explore the mechanism of Nef in DN based on miRNA regulation theory.

Methods: A DN mouse model was constructed and treated with Nef. Serum creatinine (Crea), blood urea (UREA) and urinary albumin were measured in mice by kits, and renal histopathological changes and fibrosis were observed by hematoxylin-eosin staining and Masson staining. Renal tissue superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) activities were measured by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to detect the expression of nuclear factor E2-related factor 2 (Nrf2)/ heme oxygenase 1 (HO-1) signaling pathway-related proteins in kidney tissues. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-17-5p in kidney tissues. Subsequently, a DN in vitro model was constructed by high glucose culture of human mesangial cells (HMCs), cells were transfected with miR-17-5p mimic and/or treated with Nef, and we used qRT-PCR to detect cellular miR-17 expression, flow cytometry to detect apoptosis, ELISAs to detect cellular SOD, MDA, and GSH-Px activities, Western blots to detect Nrf2/HO-1 signaling pathway-related protein expression, and dual luciferase reporter gene assays to verify the targeting relationship between Nrf2 and miR-17-5p.

Results: Administration of Nef significantly reduced the levels of blood glucose, Crea, and UREA and the expression of miR-17-5p, improved renal histopathology and fibrosis, significantly reduced MDA levels, elevated SOD and GSH-Px activities, and activated Nrf2 expression in kidney tissues from mice with DN. Nrf2 is a post-transcriptional target of miR-17-5p. In HMCs transfected with miR-17-5p mimics, the mRNA and protein levels of Nrf2 were significantly suppressed. Furthermore, miR-17-5p overexpression and Nef intervention resulted in a significant increase in high glucose-induced apoptosis and MDA levels in HMCs and a significant decrease in the protein expression of HO-1 and Nrf2.

Conclusion: Collectively, these results indicate that Nef has an ameliorative effect on DN, and the mechanism may be through the miR-17-5p/Nrf2 pathway.

目的研究奈非林(Nef)对糖尿病肾病(DN)的影响,并基于 miRNA 调控理论探讨奈非林在 DN 中的作用机制:方法:构建 DN 小鼠模型并用 Nef 治疗。用试剂盒测定小鼠血清肌酐(Crea)、血尿素(UREA)和尿白蛋白,苏木精-伊红染色和马森染色观察肾组织病理变化和纤维化。肾组织超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)活性通过酶联免疫吸附试验(ELISA)进行测定。用 Western 印迹法检测肾组织中核因子 E2 相关因子 2(Nrf2)/血红素加氧酶 1(HO-1)信号通路相关蛋白的表达。定量反转录聚合酶链反应(qRT-PCR)用于检测肾脏组织中 miR-17-5p 的表达。随后,我们通过高糖培养人间质细胞(HMCs)构建了 DN 体外模型,用 miR-17-5p mimic 和/或 Nef 处理转染细胞,并用 qRT-PCR 检测细胞 miR-17 的表达,用流式细胞术检测细胞凋亡、ELISA检测细胞SOD、MDA和GSH-Px活性,Western印迹检测Nrf2/HO-1信号通路相关蛋白的表达,以及双荧光素酶报告基因实验验证Nrf2和miR-17-5p之间的靶向关系。结果给予 Nef 能明显降低 DN 小鼠肾脏组织中的血糖、Crea、UREA 水平和 miR-17-5p 的表达,改善肾脏组织病理学和纤维化,明显降低 MDA 水平,提高 SOD 和 GSH-Px 活性,激活 Nrf2 的表达。Nrf2 是 miR-17-5p 的转录后靶标。在转染了 miR-17-5p 模拟物的 HMCs 中,Nrf2 的 mRNA 和蛋白水平均受到显著抑制。此外,miR-17-5p 过表达和 Nef 干预导致高糖诱导的 HMCs 细胞凋亡和 MDA 水平显著增加,HO-1 和 Nrf2 蛋白表达显著下降:总之,这些结果表明,Nef对DN有改善作用,其机制可能是通过miR-17-5p/Nrf2途径。
{"title":"Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis.","authors":"Huang Hongmei, Yang Maojun, L I Ting, Wang Dandan, L I Ying, Tang Xiaochi, Yuan Lu, G U Shi, X U Yong","doi":"10.19852/j.cnki.jtcm.20231204.004","DOIUrl":"10.19852/j.cnki.jtcm.20231204.004","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the effect of Neferine (Nef) on diabetic nephropathy (DN) and to explore the mechanism of Nef in DN based on miRNA regulation theory.</p><p><strong>Methods: </strong>A DN mouse model was constructed and treated with Nef. Serum creatinine (Crea), blood urea (UREA) and urinary albumin were measured in mice by kits, and renal histopathological changes and fibrosis were observed by hematoxylin-eosin staining and Masson staining. Renal tissue superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) activities were measured by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to detect the expression of nuclear factor E2-related factor 2 (Nrf2)/ heme oxygenase 1 (HO-1) signaling pathway-related proteins in kidney tissues. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-17-5p in kidney tissues. Subsequently, a DN in vitro model was constructed by high glucose culture of human mesangial cells (HMCs), cells were transfected with miR-17-5p mimic and/or treated with Nef, and we used qRT-PCR to detect cellular miR-17 expression, flow cytometry to detect apoptosis, ELISAs to detect cellular SOD, MDA, and GSH-Px activities, Western blots to detect Nrf2/HO-1 signaling pathway-related protein expression, and dual luciferase reporter gene assays to verify the targeting relationship between Nrf2 and miR-17-5p.</p><p><strong>Results: </strong>Administration of Nef significantly reduced the levels of blood glucose, Crea, and UREA and the expression of miR-17-5p, improved renal histopathology and fibrosis, significantly reduced MDA levels, elevated SOD and GSH-Px activities, and activated Nrf2 expression in kidney tissues from mice with DN. Nrf2 is a post-transcriptional target of miR-17-5p. In HMCs transfected with miR-17-5p mimics, the mRNA and protein levels of Nrf2 were significantly suppressed. Furthermore, miR-17-5p overexpression and Nef intervention resulted in a significant increase in high glucose-induced apoptosis and MDA levels in HMCs and a significant decrease in the protein expression of HO-1 and Nrf2.</p><p><strong>Conclusion: </strong>Collectively, these results indicate that Nef has an ameliorative effect on DN, and the mechanism may be through the miR-17-5p/Nrf2 pathway.</p>","PeriodicalId":94119,"journal":{"name":"Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10774715/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139428190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Efficacy of electroacupuncture on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass: a systematic review and Meta-analysis. 电针对心肺旁路心脏手术患者心肌保护和术后康复的功效:系统综述和 Meta 分析。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20230904.003
Qin Xiaoyu, Wang Chunai, Xue Jianjun, Zhang Jie, L U Xiaoting, Ding Shengshuang, G E Long, Wang Minzhen

Objective: To evaluate the efficacy of electroacupuncture (EA) intervention on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB).

Methods: Eight databases, including PubMed, Embase, the Cochrane Library, Web of Science, Chinese BioMedical Literature Database, China National Knowledge Infrastructure Database, Wanfang Data, China Science and Technology Journal Database, and two clinical trial registries, were searched. All randomized controlled trials (RCTs) related to EA intervention in cardiac surgery with CPB were collected. Based on the inclusion and exclusion criteria, two researchers independently screened articles and extracted data. After the quality evaluation, RevMan 5.3 software was used for analysis.

Results: Fourteen RCTs involving 836 patients were included. Compared with the control treatment, EA significantly increased the incidence of cardiac automatic rebeat after aortic unclamping [relative risk (RR) = 1.15, 95% confidence interval (CI) (1.01, 1.31), P < 0.05; moderate]. Twenty-four hours after aortic unclamping, EA significantly increased the superoxide dismutase [standardized mean difference (SMD) = 0.96, 95% CI(0.32, 1.61), P < 0.05; low], and interleukin (IL)-2 [SMD = 1.33, 95% CI(0.19, 2.47), P < 0.05; very low] expression levels and decreased the malondialdehyde [SMD =-1.62, 95% CI(-2.15, -1.09), P < 0.05; moderate], tumour necrosis factor-α [SMD = -1.28, 95% CI(-2.37, -0.19), P < 0.05; moderate], and cardiac troponin I [SMD = -1.09, 95% CI(-1.85, -0.32), P < 0.05; low] expression levels as well as the inotrope scores [SMD = -0.77, 95% CI(-1.22, -0.31), P < 0.05; high]. There was no difference in IL-6 and IL-10 expression levels. The amount of intraoperative sedative [SMD = -0.31, 95% CI(-0.54, -0.09), P < 0.05; moderate] and opioid analgesic [SMD = -0.96, 95% CI(-1.53, -0.38), P < 0.05; low] medication was significantly lower in the EA group than in the control group. Moreover, the postoperative tracheal intubation time [SMD = -0.92, 95% CI(-1.40, -0.45), P < 0.05; low] and intensive care unit stay [SMD = -1.71, 95% CI(-3.06, -0.36), P < 0.05; low] were significantly shorter in the EA group than in the control group. There were no differences in the time to get out of bed for the first time, total days of antibiotic use after surgery, or postoperative hospital stay. No adverse reactions related to EA were reported in any of the included studies.

Conclusions: In cardiac surgery with CPB, EA may be a safe and effective strategy to reduce myocardial ischaemia-reperfusion injury and speed up the recovery of patients after surgery. These findin

目的评估电针干预对心肺旁路(CPB)心脏手术患者心肌保护和术后康复的疗效:方法:检索了8个数据库,包括PubMed、Embase、Cochrane图书馆、Web of Science、中国生物医学文献数据库、中国国家知识基础设施数据库、万方数据、中国科技期刊数据库和两个临床试验登记处。收集了所有与 CPB 下心脏手术 EA 干预相关的随机对照试验(RCT)。根据纳入和排除标准,两名研究人员独立筛选文章并提取数据。经过质量评估后,使用RevMan 5.3软件进行分析:结果:共纳入 14 项研究,涉及 836 名患者。与对照治疗相比,EA 显著增加了主动脉瓣关闭术后心脏自动再搏的发生率[相对风险(RR)= 1.15,95% 置信区间(CI)(1.01,1.31),P 0.05;中度]。主动脉瓣关闭 24 小时后,EA 能显著提高超氧化物歧化酶 [标准化平均差 (SMD) = 0.96,95% CI(0.32, 1.61),P 0.05;低] 和白细胞介素 (IL)-2 [SMD = 1.33,95% CI(0.19, 2.47),P 0.05;极低] 的表达水平,降低丙二醛 [SMD =-1.62,95% CI(-2.15,-1.09),P 0.05;中度]、肿瘤坏死因子-α [SMD =-1.28,95% CI(-2.37,-0.19),P 0.05;中度]和心肌肌钙蛋白 I [SMD = -1.09,95% CI(-1.85,-0.32),P 0.05;低]表达水平以及肌钙蛋白评分[SMD = -0.77,95% CI(-1.22,-0.31),P 0.05;高]。IL-6和IL-10的表达水平没有差异。术中镇静剂[SMD = -0.31,95% CI(-0.54,-0.09),P 0.05;中度]和阿片类镇痛药[SMD = -0.96,95% CI(-1.53,-0.38),P 0.05;低度]的用量在EA组明显低于对照组。此外,EA 组的术后气管插管时间[SMD = -0.92,95% CI(-1.40,-0.45),P 0.05;低]和重症监护室住院时间[SMD =-1.71,95% CI(-3.06,-0.36),P 0.05;低]明显短于对照组。首次下床活动的时间、术后使用抗生素的总天数以及术后住院时间均无差异。所有纳入的研究均未报告与EA相关的不良反应:结论:在使用 CPB 的心脏手术中,EA 可能是减少心肌缺血再灌注损伤、加快患者术后恢复的一种安全有效的策略。由于大多数证据质量较低或中等,因此必须谨慎解读这些研究结果。需要更多样本量更大、质量更高的研究性试验来提供更有说服力的证据。
{"title":"Efficacy of electroacupuncture on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass: a systematic review and Meta-analysis.","authors":"Qin Xiaoyu, Wang Chunai, Xue Jianjun, Zhang Jie, L U Xiaoting, Ding Shengshuang, G E Long, Wang Minzhen","doi":"10.19852/j.cnki.jtcm.20230904.003","DOIUrl":"10.19852/j.cnki.jtcm.20230904.003","url":null,"abstract":"<p><strong>Objective: </strong>To evaluate the efficacy of electroacupuncture (EA) intervention on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB).</p><p><strong>Methods: </strong>Eight databases, including PubMed, Embase, the Cochrane Library, Web of Science, Chinese BioMedical Literature Database, China National Knowledge Infrastructure Database, Wanfang Data, China Science and Technology Journal Database, and two clinical trial registries, were searched. All randomized controlled trials (RCTs) related to EA intervention in cardiac surgery with CPB were collected. Based on the inclusion and exclusion criteria, two researchers independently screened articles and extracted data. After the quality evaluation, RevMan 5.3 software was used for analysis.</p><p><strong>Results: </strong>Fourteen RCTs involving 836 patients were included. Compared with the control treatment, EA significantly increased the incidence of cardiac automatic rebeat after aortic unclamping [relative risk (<i>RR</i>) = 1.15, 95% confidence interval (<i>CI</i>) (1.01, 1.31), <i>P <</i> 0.05; moderate]. Twenty-four hours after aortic unclamping, EA significantly increased the superoxide dismutase [standardized mean difference (<i>SMD</i>) = 0.96, 95% <i>CI</i>(0.32, 1.61), <i>P <</i> 0.05; low], and interleukin (IL)-2 [<i>SMD</i> = 1.33, 95% <i>CI</i>(0.19, 2.47), <i>P <</i> 0.05; very low] expression levels and decreased the malondialdehyde [<i>SMD</i> =-1.62, 95% <i>CI</i>(-2.15, -1.09), <i>P <</i> 0.05; moderate], tumour necrosis factor-α [<i>SMD</i> = -1.28, 95% <i>CI</i>(-2.37, -0.19), <i>P <</i> 0.05; moderate], and cardiac troponin I [SMD = -1.09, 95% <i>CI</i>(-1.85, -0.32), <i>P <</i> 0.05; low] expression levels as well as the inotrope scores [<i>SMD</i> = -0.77, 95% <i>CI</i>(-1.22, -0.31), <i>P <</i> 0.05; high]. There was no difference in IL-6 and IL-10 expression levels. The amount of intraoperative sedative [<i>SMD</i> = -0.31, 95% <i>CI</i>(-0.54, -0.09), <i>P <</i> 0.05; moderate] and opioid analgesic [<i>SMD</i> = -0.96, 95% <i>CI</i>(-1.53, -0.38), <i>P <</i> 0.05; low] medication was significantly lower in the EA group than in the control group. Moreover, the postoperative tracheal intubation time [<i>SMD</i> = -0.92, 95% <i>CI</i>(-1.40, -0.45), <i>P <</i> 0.05; low] and intensive care unit stay [<i>SMD</i> = -1.71, 95% <i>CI</i>(-3.06, -0.36), <i>P <</i> 0.05; low] were significantly shorter in the EA group than in the control group. There were no differences in the time to get out of bed for the first time, total days of antibiotic use after surgery, or postoperative hospital stay. No adverse reactions related to EA were reported in any of the included studies.</p><p><strong>Conclusions: </strong>In cardiac surgery with CPB, EA may be a safe and effective strategy to reduce myocardial ischaemia-reperfusion injury and speed up the recovery of patients after surgery. These findin","PeriodicalId":94119,"journal":{"name":"Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10774734/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139428175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Formulation, characterization and and evaluation of aloe-emodin-loaded solid dispersions for dissolution enhancement. 用于提高溶解度的芦荟大黄素负载型固体分散体的配制、表征和评估。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231110.002
L I Xiuyan, Luo Yuting, Wang Jinhui, D U Zhimin

Objective: To prepare aloe-emodin solid dispersion (AE-SD) and determine the metabolic process of AE and AE-SD in vivo.

Methods: AE-SD was prepared viasolvent evaporation or solvent melting using PEG-6000 and PVP-K30 as carriers. Thermogravimetric analysis, X-ray diffraction spectroscopy, differential scanning calorimetry, Fourier transform infrared spectroscopy and scanning electron microscopy were used to identify the physical state of AE-SD. Optimal prescriptions were screened viathe dissolution degree determination method. Using Phoenix software, AE suspension and AE-SD were subjected to a pharmacokinetic comparison study analyzing the alteration of behavior in vivo after AE was prepared as a solid dispersion. Acute toxicity was assessed in mice, and the physiological toxicity was used as the determination criterion for toxicity.

Results: AE-SD showed that AE existed in the carrier in an amorphous state. Compared with polyethylene glycol, polyvinylpyrrolidone (PVP) inhibited AE crystallization, causing the drug to transform from a dense crystalline state to an amorphous form and increasing the degree of drug dispersion. Therefore, it was more suitable as a carrier material for AE-SD. The addition of poloxamer (POL) was more beneficial to the stability of solid dispersions and could reduce the amount of PVP. The dissolution test confirmed that the optimal ratio of AE to the composite vector AE-PVP-POL was 1:2:2, and its dissolution effect was also optimal. Based on the pharmacokinetic comparison, the drug absorption was faster and quickly reached the peak of blood drug concentration in AE-SD compared to AE, the Cmax of AE-SD was greater than that of AE, and t1/2 and mean residence time of AE-SD were less than AE. The results showed that the drug metabolism in AE-SD was better, and the residence time was shorter. The toxicology study showed that both AE and AE-SD had no toxicity.

Conclusion: This paper established that the solubility of the drug could be increased after preparing a solid dispersion, as demonstrated by in vitro dissolution experiments. In vivo pharmacokinetics studies confirmed that AE-SD could improve the bioavailability of AE in vivo, providing a new concept for the research and development of AE preparations.

目的制备芦荟大黄素固体分散体(AE-SD),并测定 AE 和 AE-SD 在体内的代谢过程:方法:以 PEG-6000 和 PVP-K30 为载体,通过溶剂蒸发或溶剂熔融制备 AE-SD。采用热重分析法、X 射线衍射光谱法、差示扫描量热法、傅立叶变换红外光谱法和扫描电子显微镜鉴定 AE-SD 的物理状态。通过溶解度测定法筛选出最佳处方。使用 Phoenix 软件对 AE 悬浮液和 AE-SD 进行药代动力学比较研究,分析 AE 制成固体分散体后在体内的行为变化。对小鼠进行了急性毒性评估,并以生理毒性作为毒性的判定标准:AE-SD显示,AE以无定形状态存在于载体中。与聚乙二醇相比,聚乙烯吡咯烷酮(PVP)能抑制 AE 结晶,使药物从致密结晶状态转变为无定形状态,增加药物的分散度。因此,它更适合作为 AE-SD 的载体材料。添加聚氧乙烯(POL)更有利于固体分散体的稳定性,并可减少 PVP 的用量。溶出试验证实,AE 与复合载体 AE-PVP-POL 的最佳比例为 1:2:2,其溶出效果也是最佳的。药代动力学比较结果表明,与AE相比,AE-SD的药物吸收更快,血药浓度达峰更快,AE-SD的Cmax大于AE,AE-SD的t1/2和平均停留时间小于AE。结果表明,AE-SD 的药物代谢更好,停留时间更短。毒理学研究表明,AE 和 AE-SD 均无毒性:本文通过体外溶解实验证实,制备固体分散体后,药物的溶解度可以提高。体内药代动力学研究证实,AE-SD 可以提高 AE 在体内的生物利用度,为 AE 制剂的研究和开发提供了一个新的概念。
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引用次数: 0
B-cell lymphoma-2 phosphorylation at Ser70 site-related autophagy mediates puerarin-inhibited the apoptosis of MC3T3-E1 cells during osteoblastogenesis. B细胞淋巴瘤-2在Ser70位点的磷酸化与自噬有关,在成骨过程中介导葛根素抑制MC3T3-E1细胞的凋亡。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.20231024.002
L I Xi, Lin Xiangquan, Chen Dongdong, Liu Hui

Objective: To explore the relationship between autophagy and apoptosis regulated by puerarin during osteoblastogenesis.

Methods: In this study, the effects of puerarin on the autophagic activity and apoptosis level of osteoblast precursors (MC3T3-E1 cells) was observed. Subsequently, the roles of puerarin on B-cell lymphoma-2 (Bcl-2) phosphorylation at different sites in osteoblast precursors were observed. The effect of puerarin on the interaction between Bcl-2 and autophagy regulatory molecule or pro-apoptotic molecule was also investigated using Co-immunoprecipitation assays. In addition, the effect of puerarin on mitochondrial membrane potential of osteoblast precursors was also identified by mitochondrial membrane potential fluorescence probe assays.

Results: Our results showed that puerarin can promote the autophagic activity and apoptosis level of MC3T3-E1 cells. In addition, puerarin promoted Bcl-2 phosphorylation at Ser70 site, and the dissociation of Bcl-2-Beclin1 complex. Moreover, puerarin could enhance the binding of Bcl-2-Bcl-2-Associated X (Bax) complex in MC3T3-E1 cells. Furthermore, puerarin increased the mitochondrial membrane potential of MC3T3-E1 cells.

Conclusions: Therefore, puerarin promotes Beclin1 into autophagy flux through Bcl-2 phosphorylation at Ser70, thereby enhancing autophagy of osteoblast precursors, which mediates its anti-apoptotic role during osteoblastogenesis. Furthermore, the dissociation of Bcl-2-Beclin1 complex is conducive to the binding of Bcl-2-Bax complex, which resists the apoptosis of osteoblast precursors viathe increased mitochondrial membrane potential.

目的:探讨葛根素在成骨细胞形成过程中调控自噬和细胞凋亡的关系:方法:观察葛根素对成骨细胞前体(MC3T3-E1细胞)自噬活性和细胞凋亡水平的影响:本研究观察了葛根素对成骨细胞前体(MC3T3-E1细胞)自噬活性和凋亡水平的影响。随后,观察了葛根素对成骨细胞前体不同位点 B 细胞淋巴瘤-2(Bcl-2)磷酸化的作用。还利用共免疫沉淀实验研究了葛根素对 Bcl-2 与自噬调控分子或促凋亡分子之间相互作用的影响。此外,葛根素对成骨细胞前体线粒体膜电位的影响也通过线粒体膜电位荧光探针测定得以确定:结果:葛根素能促进MC3T3-E1细胞的自噬活性和凋亡水平。此外,葛根素还能促进Bcl-2在Ser70位点的磷酸化和Bcl-2-Beclin1复合物的解离。此外,葛根素还能增强 MC3T3-E1 细胞中 Bcl-2-Bcl-2-Associated X(Bax)复合物的结合。此外,葛根素还能提高MC3T3-E1细胞的线粒体膜电位:因此,葛根素通过Bcl-2在Ser70处的磷酸化促进Beclin1进入自噬通路,从而增强成骨细胞前体的自噬,这是其在成骨过程中抗凋亡作用的介导因素。此外,Bcl-2-Beclin1 复合物的解离有利于 Bcl-2-Bax 复合物的结合,而 Bcl-2-Bax 复合物可通过线粒体膜电位的增加抵御成骨细胞前体的凋亡。
{"title":"B-cell lymphoma-2 phosphorylation at Ser70 site-related autophagy mediates puerarin-inhibited the apoptosis of MC3T3-E1 cells during osteoblastogenesis.","authors":"L I Xi, Lin Xiangquan, Chen Dongdong, Liu Hui","doi":"10.19852/j.cnki.jtcm.20231024.002","DOIUrl":"10.19852/j.cnki.jtcm.20231024.002","url":null,"abstract":"<p><strong>Objective: </strong>To explore the relationship between autophagy and apoptosis regulated by puerarin during osteoblastogenesis.</p><p><strong>Methods: </strong>In this study, the effects of puerarin on the autophagic activity and apoptosis level of osteoblast precursors (MC3T3-E1 cells) was observed. Subsequently, the roles of puerarin on B-cell lymphoma-2 (Bcl-2) phosphorylation at different sites in osteoblast precursors were observed. The effect of puerarin on the interaction between Bcl-2 and autophagy regulatory molecule or pro-apoptotic molecule was also investigated using Co-immunoprecipitation assays. In addition, the effect of puerarin on mitochondrial membrane potential of osteoblast precursors was also identified by mitochondrial membrane potential fluorescence probe assays.</p><p><strong>Results: </strong>Our results showed that puerarin can promote the autophagic activity and apoptosis level of MC3T3-E1 cells. In addition, puerarin promoted Bcl-2 phosphorylation at Ser70 site, and the dissociation of Bcl-2-Beclin1 complex. Moreover, puerarin could enhance the binding of Bcl-2-Bcl-2-Associated X (Bax) complex in MC3T3-E1 cells. Furthermore, puerarin increased the mitochondrial membrane potential of MC3T3-E1 cells.</p><p><strong>Conclusions: </strong>Therefore, puerarin promotes Beclin1 into autophagy flux through Bcl-2 phosphorylation at Ser70, thereby enhancing autophagy of osteoblast precursors, which mediates its anti-apoptotic role during osteoblastogenesis. Furthermore, the dissociation of Bcl-2-Beclin1 complex is conducive to the binding of Bcl-2-Bax complex, which resists the apoptosis of osteoblast precursors <i>via</i>the increased mitochondrial membrane potential.</p>","PeriodicalId":94119,"journal":{"name":"Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10774730/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139428156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Effects of anterior sciatic nerve acupuncture on lower limb paralysis after cerebral infarction: study protocol for a randomized controlled trial. 坐骨神经前叶针刺对脑梗塞后下肢瘫痪的影响:随机对照试验研究方案。
Pub Date : 2024-02-01 DOI: 10.19852/j.cnki.jtcm.2024.01.001
L I Menghan, Wang Yu, Ran Dawei, Yang Xinming, Deng Shizhe, Shi Lei, Meng Zhihong

Stroke is the main cause of disability in the middle and old age. Hemiplegia, especially lower limb paralysis, often leads to the loss of self-care ability and a series of secondary injuries. The main method to improve hemiplegic limb movement is exercise therapy, but there are still many patients with disabilities after rehabilitation treatment. As one of the non-pharmacological therapies for stroke, acupuncture has been recognized to improve motor function in patients. Here, we propose a new method, anterior sciatic nerve acupuncture, which can stimulate both the femoral nerve and the sciatic nerve. We designed this study to determine the effect of this method on lower limb motor function. Sixty participants recruited with hemiplegia after cerebral infarction will be randomly assigned to the test group or control group in a 1:1 ratio. The control group will receive Xingnao Kaiqiao acupuncture, and the test group will receive anterior sciatic nerve acupuncture on this basis. All participants will get acupuncture treatment once a day, 6 times a week for 2 weeks. The primary outcome is Fugl-Meyer Assessment of Lower Extremity and the secondary outcomes are Modified Ashworth Scale and Modified Barthel Index. Data will be collected before treatment, 1 week after treatment, and 2 weeks after treatment, and then statistical analysis will be performed. This study can preliminarily verify the effect of anterior sciatic nerve acupuncture on improving lower limb motor function in patients with cerebral infarction, which may provide an alternative approach for clinical treatment of hemiplegia.

中风是中老年致残的主要原因。偏瘫,尤其是下肢瘫痪,往往导致生活自理能力丧失,并引发一系列继发性损伤。运动疗法是改善偏瘫肢体运动的主要方法,但康复治疗后仍有不少患者致残。针灸作为治疗中风的非药物疗法之一,其改善患者运动功能的作用已得到认可。在此,我们提出了一种新方法--坐骨神经前针刺法,它可以同时刺激股神经和坐骨神经。我们设计了这项研究,以确定这种方法对下肢运动功能的影响。我们将招募 60 名脑梗塞偏瘫患者,按 1:1 的比例随机分配到试验组和对照组。对照组将接受星脉开窍针刺,试验组将在此基础上接受坐骨神经前神经针刺。所有参与者每天接受一次针灸治疗,每周 6 次,持续 2 周。主要结果为 Fugl-Meyer 下肢评估,次要结果为改良阿什沃斯量表和改良巴特尔指数。将收集治疗前、治疗后 1 周和治疗后 2 周的数据,然后进行统计分析。本研究可初步验证针刺坐骨神经前束对改善脑梗死患者下肢运动功能的作用,为临床治疗偏瘫提供了一种可供选择的方法。
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Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan
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