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Article in Chinese | WPRIM | ID: wpr-905992


Objective:To investigate the possible mechanism of Xieheyin in alleviating obese polycystic ovary syndrome with insulin resistance(PCOS-IR)and reducing inflammatory response. Method:Ten of sixty SPF femlae C57BL/6J mice were randomly selected as the normal group,and the rest mice were given letrozole 0.002 g·kg<sup>-1</sup> combined with fecal suspension 2 g·kg<sup>-1</sup> for 28 consecutive days to establish model of PCOS-IR.The mice that were successfully modeled were randomized into the model group,metformin group(0.25 g·kg<sup>-1</sup>),and low(10 g·kg<sup>-1</sup>),medium(20 g·kg<sup>-1</sup>),and high-dose(40 g·kg<sup>-1</sup>)Xieheyin groups,and administered with the corresponding drugs by gavage,once a day,for four consecutive weeks. Except the normal control group, the mice in the other groups were continuously given fecal suspension combined with letrozole solution to maintain the model during the treatment. The mice were weighed once a week.Levels of fasting blood glucose (FBG) were detected by blood glucose test strips.And enzyme-linked immunosorbent assay (ELISA) method was used to detect serum testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),fasting insulin(FINS)level,and LH/FSH and Homeostasis model assesment of insulin resistance (HOMA-IR) were calculated.The uterus and ovaries were weighed and fixed.Hematoxylin-eosin(HE)staining was used to observe ovarian tissue pathology morphology. Western blot was used to detect the expression levels of tight junction key molecular zonula occludens 1(ZO-1),occludin in colon tissues,and the expression levels of Toll-like receptor 4/nuclear factor kappa B/Nod-like receptor protein 3(TLR4/NF-<italic>κ</italic>B/NLRP3)signaling pathway and inflammation associated proteins cysteinyl aspartate specific proteinase-1(Caspase-1) and interleukin-1<italic>β</italic>(IL-1<italic>β</italic>) in colon tissues. Result:Compared with normal control group,the body weight of mice in the model control group increased significantly(<italic>P</italic><0.01). Serum FINS,FBG,HOMA-IR,T,LH/FSH were significantly increased(<italic>P</italic><0.05,<italic>P</italic><0.01). The uterine organ ratio were decreased significantly(<italic>P</italic><0.01),while the ovarian organ ratio were significantly increased(<italic>P</italic><0.01). The number of atresia follicles and cystic dilatation follicles increased significantly,and the number of corpus luteum significantly decreased,the thickness of follicular granulosa cells also decreased,while the white membrane thickness of the ovary increased. Tight junction related ZO-1,occludin proteins in colon tissues were all decreased(<italic>P</italic><0.01).The relative expression levels of inflammation-related protein IL-1<italic>β</italic>,Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein signaling pathway were significantly increased(<italic>P</italic><0.05).Compared with model control group, the body weight of mice in the low,middle and high dose Xieheyin group decreased significantly(<italic>P</italic><0.01). The serum T,LH/FSH,FINS,FBG,HOMA-IR were significantly decreased(<italic>P</italic><0.05,<italic>P</italic><0.01). The uterine organ ratio were increased(<italic>P</italic><0.05),while the ovarian organ ratio were decreased(<italic>P</italic><0.05). The number of cystic follicles decreased and corpus luteum increased,the thickness of follicular granulosa cells increased and be arranged normally,while the white membrane thickness of the ovary increased slightly. The expressions of ZO-1,occludin proteins were increased(<italic>P</italic><0.01). The expression levels of IL-1<italic>β</italic>,Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein in the high dose group were significantly decreased(<italic>P</italic><0.01). Conclusion:Xieheyin could activate intestinal TLR4/NF-<italic>κ</italic>B/NLRP3 signaling pathway,inhibit pro-inflammatory factor secretion,improve obesity and IR,which was correlated with rebuilding intestinal mucosal barrier and inhibiting intestinal inflammation.

Article in Chinese | WPRIM | ID: wpr-878876


The effect of Shouhui Tongbian Capsules(SHTB) on the endogenous metabolites of colon tissue in mice with slow transit constipation was analyzed by metabolomics methods to explore its mechanism in the treatment of constipation. ICR mice were randomly divided into normal group, model group and SHTB group according to the body weight. The mice were given diphenoxylate to establish the slow transit constipation model. Mouse carbon ink pushing rate, first defecation time and the number of defecation particles in 12 h were observed. The mouse colon tissue was separated and the mucous cells were detected by Periodic acid Schiff and Alcian blue(AB-PAS) staining. Ultra-high-performance liquid chromatography electrospray ionization orbitrap tandem mass spectrometry(UPLC-ESI-Orbitrap-MS/MS) technology was used to characterize the differences in tissue metabolism to screen out the potential different metabolites and possible metabolic pathways in colon tissue. The results indicated that SHTB could significantly shorten the first defecation time and the number of defecations, and increase the number of intestinal peristalsis and mucous cells in the colonic mucosa compared to the model mice. Metabolomics results showed that, compared with the normal group, a total of 17 potential biomarkers, including L-kynurenine, N6,N6,N6-trimethyl-L-lysine, L-formylkynurenine, N6-acetyl-L-lysine, L-phenylalanine, phenylacetaldehyde, xanthoxin, thymidine, glycyl-L-leucine, cystathionine,(R)-1-aminopropan-2-ol, deoxycytidine, gamma-glutamyl-gamma-aminobutyraldehyde, D-galactose, L-arginine, L-proline and pyruvate, were found and identified in colon tissue. Treated with SHTB, these metabolic differences tended to return to normal levels. Therefore, it could be made a conclusion that the therapeutic effect of SHTB on chronic transit constipation may be related to regulating phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arginine and proline metabolism, cysteine and methionine metabolism, tyrosine metabolism, arginine biosynthesis, pyruvate metabolism, glycolysis, pyrimidine metabolism, tricarboxylic acid cycle and galactose metabolism.

Animals , Biomarkers , Capsules , Chromatography, High Pressure Liquid , Constipation/drug therapy , Metabolomics , Mice , Mice, Inbred ICR , Tandem Mass Spectrometry
Article in Chinese | WPRIM | ID: wpr-878873


To explore the mechanism of Shouhui Tongbian Capsules in treating constipation by means of network pharmacology and molecular docking approach. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and Bioinfoematics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN) were applied to obtain chemical components and potential targets of eight herbs in Shouhui Tongbian Capsules according to the screening principles of oral availability(OB)≥30% and drug-like property(DL)≥0.18. Disease targets relating to constipation were screened out through GeneCards, PharmGkb and other databases, drug targets were integrated with disease targets, and intersection targets were exactly the potential action targets of Shouhui Tongbian Capsules for treating constipation; PPI network of potential targets was constructed using STRING platform, and GO(gene ontology) analysis and KEGG(Kyoto encyclopedia of genes and genomes) pathway data were obtained to conduct enrichment analysis and predict its mechanism of action. Cytoscape 3.6.1 was used to construct a network of "medicinal materials-chemical components-drug targets", and the network topology analysis was carried out on the PPI network to obtain its main components and key targets. Molecular docking between components and key targets of Shouhui Tongbian Capsules verified the accuracy of network pharmacological analysis results. The PPI network analysis showed 92 chemical components, including quercetin, stigmaste-rol, aloe-emodin, rhein, and key targets for instance AKT1, MAPK1, IL6, JUN, TNF and TP53. The enrichment analysis of KEGG screened out 157 signal pathways(P<0.01), mainly involving interleukin 17 signaling pathway, AGE-RAGE signaling pathway in diabetic complications, thyroid hormone signaling pathway. Quercetin, resveratrol and lysine with top degree value had a rational conformation in docking site of protein crystal complexes. This study preliminarily showed that various active ingredients in Shouhui Tongbian Capsules could regulate multiple signaling pathways, increase intestinal smoothness and peristalsis function, ensure smooth intestinal lumen, and play a role in treating constipation by acting on key targets, such as AKT1, MAPK1, IL6 and JUN.

Capsules , Constipation/genetics , Drugs, Chinese Herbal , Humans , Medicine, Chinese Traditional , Molecular Docking Simulation
Article in Chinese | WPRIM | ID: wpr-878872


"Target fishing" strategy was used to investigate the direct targets and mechanism of Shouhui Tongbian Capsules on relaxing bowel. Magnetic beads cross-linked with the chemical constituents from Shouhui Tongbian Capsules were prepared. The potential target proteins were captured from the total protein lysates of rat intestine using the beads. The captured proteins were further identified by LC-MS/MS, and the associated pathways were analyzed by Cytoscape. RESULTS:: showed that 138 potential target proteins were identified, which were involved in eight signaling pathways, including tricarboxylic acid cycle, pyrimidine metabolism, sulfur metabolism, fatty acid degradation, alanine/aspartate/glutamate metabolism, arginine/proline metabolism, valine/leucine/isoleucine degradation, and β-alanine metabolism. Taken together, Shouhui Tongbian Capsules may exert relaxing bowel effect by acting on multiple signaling pathways to promote intestinal gurgling, inhibit inflammation, as well as improve intestinal barrier function, intestinal water secretion, and intestinal flora.

Animals , Capsules , Chromatography, Liquid , Intestines , Leucine , Rats , Tandem Mass Spectrometry
Acta Pharmaceutica Sinica ; (12): 482-489, 2014.
Article in English | WPRIM | ID: wpr-245058


This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.

Animals , Anti-Inflammatory Agents , Pharmacology , Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Arctium , Chemistry , Cell Cycle Checkpoints , Cell Proliferation , Cytokines , Metabolism , Female , Furans , Pharmacology , Interferon-gamma , Metabolism , Interleukin-10 , Metabolism , Interleukin-2 , Metabolism , Interleukin-2 Receptor alpha Subunit , Metabolism , Interleukin-4 , Metabolism , Interleukin-6 , Metabolism , Ionomycin , Pharmacology , Lectins, C-Type , Metabolism , Lignans , Pharmacology , Lymphocyte Activation , Mice , Mice, Inbred BALB C , Plants, Medicinal , Chemistry , T-Lymphocytes , Cell Biology , Allergy and Immunology , Tetradecanoylphorbol Acetate , Pharmacology , Tumor Necrosis Factor-alpha , Metabolism