Zhongguo Zhongyao Zazhi最新文献

The chemical constituents of the roots of Berberis polyantha were investigated by various chromatographic separation methods, including silica gel column chromatography, SP825 column chromatography, polyamide column chromatography, reversed-phase C_(18) column chromatography, and preparative high-performance liquid chromatography. The structures of compounds were identified through physicochemical properties and spectroscopic techniques(1D NMR, 2D NMR, and X-ray). From the methanol extract of the roots of B. polyantha, six compounds were isolated and identified as polyanthaine(1), corydaldine(2), noroxyhydrastinine(3), 8-oxypalmatine(4), 8-oxyberberine(5), and rugosinone(6). Compound 1 was a novel compound and named polyanthaine, and other compounds were isolated from this plant for the first time. The anti-inflammatory effect of the compounds was evaluated by measuring the release of nitric oxide(NO) in the culture of lipopolysaccharide(LPS)-induced RAW264.7 macrophages. At a concentration of 10 μmol·L~(-1), compound 1-3,5,6 demonstrated inhibitory activity on the release of NO in LPS-induced RAW264.7 cells.

Atractylodis Macrocephalae Rhizoma(Baizhu), the dried rhizome of Atractylodes macrocephala in the Compositae family, was first recorded in the Shen Nong's Materia Medica(Shen Nong Ben Cao Jing). It is warm in nature, sweet and bitter in taste, and belongs to the spleen and stomach meridians. Baizhu exerts effects of strengthening the spleen and benefiting Qi, drying dampness and promoting diuresis. As a core drug in the traditional Chinese medicine(TCM) theory of "strengthening the spleen and regulating the bowels", its mechanism in treating constipation goes beyond the traditional single perspective of "strengthening the spleen and moistening the intestines", reflecting instead a systematic regulation of intestinal homeostasis. TCM holds that Baizhu tonifies the middle energizer and regulates Qi movement, restoring the physiological function of "the spleen governing the transportation of body fluids for the stomach", thereby fundamentally resolving intestinal transmission disorders caused by spleen deficiency, Qi stagnation, and body fluid imbalance. Modern research has confirmed that the main active components of Baizhu include volatile oils, polysaccharides, and lactones. Its mechanism in treating constipation exhibits multi-target synergy: regulating intestinal flora diversity, promoting the synthesis of short-chain fatty acids(SCFAs), and enhancing intestinal mucosal barrier function; activating the 5-hydroxytryptamine receptor 4(5-HT4) pathway of interstitial cells of Cajal(ICC) in the intestinal nerve plexus, enhancing the rhythmic contraction of intestinal smooth muscle, and significantly improving intestinal motility; protecting the intestinal mucosal barrier and promoting mucus secretion; regulating the expression of aquaporins(AQPs) and increasing fecal water content. In compound prescriptions, Baizhu exerts synergistic and enhancing effects guided by the clinical application: combined with Yin-nourishing drugs, it can enhance mucus secretion; combined with Qi-moving drugs, it can improve intestinal mechanical motility, forming a formula strategy that treats both symptoms and root causes. This article systematically analyzes the integrated effects of Baizhu on constipation from a three-dimensional perspective of "components-mechanisms-compatibility", providing new insights for developing precise therapies for constipation based on Baizhu, and demonstrating the application value of TCM in modern chronic disease management.

Research has demonstrated that the O-methylation of secondary metabolites is mainly catalyzed by O-methyltransferases(OMTs). The oxygen-methylated flavonoids in plants of Asteraceae are the most abundant. However, the evolutionary pathway of the new biosynthetic capacity of flavonoids in Chrysanthemum indicum remains unknown. In this study, the methyltransferase gene CiCOMT27 was cloned from C. indicum and expressed in Escherichia coli, and the recombinant protein was isolated and purified by column chromatography. UPLC analysis showed that CiCOMT27 could transfer the methyl group from S-adenosyl-L-methionine(SAM) to the 4'-OH of luteolin and quercetin to form diosmetin and tamarixetin, and to the 3'-OH of eriodictyol to form homoeriodictyol. In addition, CiCOMT27 could catalyze the 7-OH methylation of hispidulin and scutellarein to form their corresponding methoxy derivatives, indicating that CiCOMT27 had the function of catalyzing methylation at multiple sites. When luteolin was used as the substrate, the conversion rate was as high as over 80%, and when eriodictyol, hispidulin, and scutellarein were used as substrates, the conversion rate could also reach about 60%. In addition, molecular docking experiments were conducted between CiCOMT27 and the physiological substrate luteolin of C. indicum. The results showed that luteolin was more likely to bind to CiCOMT27 at the 4'-OH site, which supported at the structural level that the CiCOMT27 had a significant preference for flavonoid substrates with hydroxyl substituents. This study provides a new molecular mechanism explanation for the structural diversity of flavonoids in C. indicum and lays a foundation for the directed design of specific methylated flavonoid derivatives.

Based on the model of spleen deficiency, this study investigated the medicinal properties of spleen meridian tropism of Ganoderma lucidum spores and their efficacy of strengthening the spleen. Fifty SPF Kunming mice were randomly divided into a control group, a model group, an Atractylodes macrocephala group(1.1 g·kg~(-1)), a low-dose G. lucidum spores group(0.55 g·kg~(-1)), and a high-dose G. lucidum spores group(1.1 g·kg~(-1)). The mouse spleen deficiency model was established by fasting every other day, having adequate feed on the other days, and performing exhausted swimming every day for 21 consecutive days. From the first day of modeling, deionized water was given to mice in the control group and the model group in the morning of each day, and the corresponding medicine was given to the rest of the groups, with the medicines administered for 21 consecutive days. The physical signs and behavior, digestive function, energy metabolism, and immune regulation related symptoms of the mice were detected. The results showed that G. lucidum spores could improve the symptoms of spleen deficiency in mice, such as emaciation, lack of luster of fur, reduced food intake, huddling and laziness, make stools shaped and activity increased, enhance the body mass of the mice, increase their grip strength, and prolong the time of exhausted swimming. G. lucidum spores significantly increased serum D-xylose level, enhanced motilin(MTL) and gastrin(GAS) secretion, and decreased cholecystokinin(CCK), vasoactive intestinal peptide(VIP), and somatostatin(SST) mRNA expression levels in mice with spleen deficiency syndrome. G. lucidum spores elevated the levels of hepatic glycogen(LG), muscle glycogen(MG), and adenosine triphosphate(ATP), enhanced the activities of Na~+-K~+-ATPase, Ca~(2+)-Mg~(2+)-ATPase, lactate dehydrogenase(LDH), mitochondrial respiratory enzyme Ⅰ, mitochondrial respiratory enzyme Ⅳ, and ATPase, and reduced the accumulation of free fatty acid(FFA) and lactic acid(LA). G. lucidum spores increased thymus index, spleen index, elevated immunoglobulin A(IgA), immunoglobulin M(IgM), and immunoglobulin G(IgG) levels, decreased interleukin-6(IL-6), interleukin-1 beta(IL-1β), and tumor necrosis factor-α(TNF-α) levels, and improved splenic histopathological changes. The above results indicate that G. lucidum spores can improve the physical signs and behavior, digestive function, energy metabolism, and immune regulation of mice with spleen deficiency and have the medicinal properties of spleen meridian tropism and efficacy of strengthening the spleen.

This study explores whether Angelicae Sinensis Radix extract alleviates fibrosis of human fibroblast-like synoviocytes(FLSs) by regulating the histone H3 lysine 18 lactylation(H3K18la)/methyltransferase-like protein 3(METTL3)/N~6-methyladenosine(m~6A) axis-mediated m~6A methylation of Netrin-1. FLSs were grouped as follows: control, transforming growth factor(TGF)-β1, TGF-β1+Angelicae Sinensis Radix extract drug-containing serum(AS), and TGF-β1+AS+sodium lactate(Nala). Other groups except the control group were stimulated with 10 ng·mL~(-1) TGF-β1 for 24 h for the modeling of knee osteoarthritis(KOA)-like fibrosis. The TGF-β1+AS group was treated with 10% AS-containing serum for 24 h, while the TGF-β1+AS+Nala group received a combination of AS and 5 mmol·L~(-1) Nala(a METTL3 inhibitor) for 24 h. Following the interventions, hematoxylin-eosin(HE) staining and Masson's trichrome staining were performed to evaluate the effects of AS on cellular morphology and collagen deposition. Western blot was performed to analyze the levels of pan-Kla, H3K18la, METTL3, inflammatory cytokines [interleukin-6(IL-6), tumor necrosis factor-alpha(TNF-α), and interleukin-1 beta(IL-1β)], and fibrosis-related proteins [collagen Ⅰ, alpha-smooth muscle actin(α-SMA), tissue inhibitor of matrix metalloproteinase-1(TIMP-1), and vimentin]. Immunofluorescence staining was employed to measure the levels of H3K18la and α-SMA, and qPCR was employed to measure the mRNA level of METTL3. ELISA quantified the levels of IL-6, TNF-α, and IL-1β in supernatants. Online tools were used to predict the m~6A methylation sites on Netrin-1. Lentiviral transfection was used to knock down METTL3(sh-METTL3) and blank plasmid(sh-NC) or overexpress METTL3(oe-METTL3), Netrin-1(oe-Netrin-1), and blank plasmid(oe-NC). RNA immunoprecipitation sequencing(RIP) was conducted to study the mRNA interaction between METTL3 and Netrin-1. MeRIP-qPCR was employed to assess the m~6A level of Netrin-1, while qPCR and Western blot were employed to evaluate the mRNA and protein levels, respectively, of METTL3 and Netrin-1. Compared with the control group, the TGF-β1 group exhibited significantly elevated levels of pan-Kla, H3K18la, METTL3, IL-6, TNF-α, IL-1β, collagen Ⅰ, α-SMA, TIMP-1, and vimentin. AS reduced the levels of these factors, while the TGF-β1+AS+Nala group showed increased levels compared with the TGF-β1+AS group. Netrin-1 contained multiple m~6A methylation sites. Compared with the IgG control group, sh-METTL3 lowered the levels of Netrin-1 and m~6A-modified Netrin-1, whereas oe-METTL3 increased the levels of Netrin-1 and m~6A-modified Netrin-1. In addition, sh-METTL3 downregulated the mRNA/protein levels of METTL3 and Netrin-1. AS regulates the H3K18la/METTL3/m~6A axis to inhibit the m~6A methylation of Netrin-1, reduce inflammatory cytokine secretion, and down-regulate fibrosis-related gene expression, thereby alleviating synovial fibrosis in KOA.

This study aims to explore the effects and mechanisms of the core formula(TCF) improving glomerulosclerosis(GS) in diabetic kidney disease(DKD) by Jinling medical school based on reticulophagy(ER-phagy)-pyroptosis-apoptosis-necroptosis(PANoptosis) signaling pathway in podocytes. Firstly, the modified DKD rat models were established. After modeling successfully, the DKD rat models received either TCF, empagliflozin(EMPA), or saline by gavage for 8 weeks, respectively. Secondly, MPC5 cells were subjected to treatment under high-glucose(HG) conditions alone, or HG combined individually with drug-containing serum of the low-dose of TCF(L-TCF) and the high-dose of TCF(H-TCF), EMPA, or Torin 1(autophagy agonist). In the in vivo studies, the changes of general conditions and renal injury indicators, pathological characteristics of GS, and the protein expression levels of podocyte marker molecules were observed, respectively. Furthermore, the changes of the level of reactive oxygen species(ROS) in the kidneys, the expression levels of the core receptor and marker molecules of ER-phagy, and the expression degree of Z-DNA binding protein 1(ZBP1) were also observed. In the in vivo and in vitro studies, we analyzed changes in the expression levels of ZBP1 protein in the kidneys and podocytes, as well as the protein expression levels of key signaling molecules in the cellular PANoptosis were investigated, respectively. The in vivo study results showed that, for the DKD model rats, general conditions and renal injury indicators, pathological characteristics of GS, and the protein expression levels of podocyte marker molecules were improved, respectively. Moreover, the level of ROS in the kidneys, the expression level of the core receptor and marker molecules of ER-phagy, and the expression degree and protein expression level of ZBP1, as well as the protein expression levels of key signaling molecules in the cellular PANoptosis were ameliorated, respectively. Notably, the improvement effects of TCF and EMPA were basically similar, but the beneficial effect of the protein expression level of interleukin-18(IL-18) in the kidneys, as an inflammatory factor, was superior to EMPA. The in vitro study results showed that, high and low doses of TCF and EMPA both could ameliorate the protein expression levels of key signaling molecules in the PANoptosis pathway in podocytes treated with HG, and the improvement effect of H-TCF was superior to that of L-TCF or EMPA. In conclusion, the study clarifies that TCF improves GS in DKD by regulating ER-phagy-PANoptosis signaling pathway in podocytes in vivo and in vitro, and has laid the groundwork for exploring mechanisms and scientific implications of treating diabetic kidney disease by Jinling medical school.

This study aimed to explore the molecular mechanism underlying the protective effect of Danggui Buxue Decoction(DBD) against myocardial injury induced by chronic intermittent hypoxia(CIH). A CIH model was established in Sprague-Dawley(SD) rats, and DBD was administered via intragastric gavage at low, medium, and high doses(3.6, 7.2, and 14.4 g·kg~(-1)·d~(-1)). An intermittent hypoxia(IH) model was established in H9c2 cells. Cells were treated with DBD-containing serum, DBD-containing serum combined with the AMPK inhibitor Compound C, or the AMPK activator A769662. Molecular docking was used to analyze the binding affinity and interaction between the main active components of DBD and the AMPK protein. Cardiac function, myocardial histopathological changes, and mitochondrial morphological alterations were detected in SD rats. In myocardial tissue and H9c2 cells, mitochondrial membrane potential(JC-1), the activity of mitochondrial respiratory complexes Ⅰ and Ⅱ, ATP content, and MDC fluorescence staining intensity were measured. The expression levels of mitophagy-related proteins(Beclin-1, LC3-Ⅱ, LC3-Ⅰ, Parkin, PINK1, P62), apoptosis-related proteins(Bcl-2, BAX, cleaved caspase-3, caspase-3, p-p70S6K, p70S6K, Cyt C), and proteins related to the AMPK/mTOR signaling pathway(p-AMPK, AMPK, p-mTOR, mTOR, p-ULK1, ULK1) were also detected. The results showed that DBD significantly improved cardiac function and alleviated myocardial injury in CIH rats. DBD and its medicated serum upregulated the expression of Beclin-1, LC3-Ⅱ/LC3-Ⅰ, Parkin, PINK1, Bcl-2/BAX, mitochondrial Cyt C, p-AMPK/AMPK, and p-ULK1/ULK1, while downregulating the expression of P62, cleaved caspase-3/caspase-3, and p-p70S6K/p70S6K, p-mTOR/mTOR. Additionally, DBD increased mitochondrial membrane potential, the activity of respiratory complexes I and Ⅱ, ATP levels, and MDC fluorescence intensity. The AMPK activator A769662 exerted therapeutic effects similar to those of DBD, whereas the AMPK inhibitor compound C significantly suppressed the therapeutic effect of DBD-containing serum. These findings suggest that DBD alleviates CIH-induced myocardial injury by activating the AMPK/mTOR signaling pathway, thereby promoting mitophagy and inhibiting mitochondria-mediated apoptosis.

To rapidly evaluate Ziziphi Spinosae Semen samples with different frying degrees, this study employed a CM-5 spectrophotometer to detect the dynamic color changes during the frying process. Near-infrared spectroscopy was utilized to measure Ziziphi Spinosae Semen samples at different frying degrees via diffuse reflectance. Principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were combined to quickly assess the quality of Ziziphi Spinosae Semen at different frying degrees. The results showed that with prolonged frying time, the brightness of Ziziphi Spinosae Semen decreased slightly, while the red and yellow hues became more pronounced, and color saturation increased. The saturation value(C~*), yellow-blue value(b~*), and red-green value(a~*) were the primary variables contributing to the color difference between raw and fried Ziziphi Spinosae Semen. The OPLS-DA model was employed to distinguish between raw and fried Ziziphi Spinosae Semen based on color parameters. The near-infrared raw spectra of Ziziphi Spinosae Semen samples with varying frying degrees were pretreated and optimized using multiple scatter correction(MSC) and competitive adaptive reweighted sampling(CARS). The PLS-DA model effectively differentiated samples with different frying degrees. The R~2 values for both the calibration and prediction sets of the discrimination model were greater than 0.90, indicating good discriminative capability. This study achieved rapid evaluation of different frying degrees of Ziziphi Spinosae Semen decoction pieces and provides a scientific reference for quality control during the processing of fried Ziziphi Spinosae Semen decoction pieces.