Based On The InsR-PI3K-Akt Signaling Pathway To Explore The Mechanism Of Jinqi Jiangtang Tablet Improving Insulin Resistance In Cells

ObjectiveJinqi Jiangtang Tablet is a commonly used Chinese patent medicine with good effect in the treatment of type 2 diabetes.However,due to the lack of clinical trials and pharmacodynamic mechanism research,the material basis and mechanism of its hypoglycemic effect are still unclear.Therefore,this paper uses the network meta-analysis method to evaluate the clinical efficacy of Jinqi Jiangtang tablets,analyzes the potential targets and target pathways of Jinqi Jiangtang tablets in improving insulin resistance in patients with type 2 diabetes from the perspective of network pharmacology,and then establishes the cell model of islet resistance by cell experiment in vitro to verify the mechanism of Jinqi Jiangtang tablets in improving insulin resistance.Methods1.Retrieves PubMed,Web of Science,Cochrane Library,Embase,China National Knowledge Infrastructure(CNKI),WanFang Datebase,China Biomedical Document Service System Database(SinoMed),Chinese Science and Technology Periodical Database(VIP)About Jinqi Jiangtang tablets and Other Chinese medicine treatment of 2 diabetes Separate application or Combined with Metformin.Filter the literature in advance and extract related information,and extracts the risk bias evaluation.Based on Bayesian theory,a mesh relationship diagram,a funnel map,a league map,etc.According to the surface under the cumulative ranking area(SUCRA)and compared the effectivenessand,sorted according to the results of the curve.2.Based on Network Pharmacology Technologies Gets the component structure in the TCMSP database and the PubChem database and imports the Swiss Target Prediction database to obtain the ingredient target.The disease target was obtained in OMIM and genecards database,and Wayne map was drawn.The drug disease common target was obtained by intersection of the two databases.Bring the common target into the Cytoscape Software to construct a "drug-component-target-disease" network relationship of Jinqi Jiangtang Tablet.Using the NetWork Analyzer function to topology the main active ingredients to obtain the core ingredient rankings.Constructing a protein interaction relationship network diagram(PPI),drawing a core target sorting diagram.Based on R software to perform a key target gene of GO and KEGG enrichment analysis,a possible pathway that plays a role in the treatment of type 2 diabetic insulin resistance.3.HepG2 cell line of hepatoblastoma was selected to establish insulin resistance model by high glucose combined with insulin long-term stimulation,and cell counting kit 8(CCK8)method was used to detect the effect of modeling on cell activity.The HepG2 cell model was divided into normal cells,model cells and Jinqi Jiangtang tablets intervention group.The expression level,phosphorylation level and mRNA expression level of key proteins in the pathway were analyzed by real time PCR(RT-PCR)and Western blot(WB)respectively.Results1.In the network meta-analysis,71 RCTs involving 6882 patients and 9 interventions were included.The results of network meta-analysis showed that in the aspect of combination of traditional Chinese medicine and Western medicine,the efficacy of combination of five kinds of traditional Chinese medicine and Western medicine was significantly higher than that of Western medicine control group,and in the aspect of reducing 2hPG and HbAlc,the efficacy of Jinqi Jiangtang tablets combined with metformin was better than that of other four kinds of traditional Chinese medicine.In terms of reducing FBG,the comparison results of single and combined use of traditional Chinese medicine and metformin were as follows:Gegen Qinlian Decoction+metformin>Jinlida+metformin>Major Bupleurum Decoction+metformin>Gegen Qinlian Decoction>Jinqi Jiangtang+metformin>Jinlida>Major Bupleurum Decoction>Tianqi Jiangtang+metformin>metformin.In terms of reducing 2hPG,the comparison results were as follows:Gegen Qinlian Decoction>Jinqi Jiangtang+metformin>Gegen Qinlian Decoction+metformin>Jinlida+metformin>Tianqi Jiangtang+metformin>Major Bupleurum Decoction+metformin>Jinlida>Jinqi Jiangtang>metformin>Major Bupleurum decoction.In terms of reducing HbAlc,the comparison results were as follows:Jinqi Jiangtang+metformin>Gegen Qinlian Decoction+metformin>Major Bupleurum Decoction+metformin>Gegen Qinlian Decoction>Jinlida+metformin>Tianqi Jiangtang+metformin>Jinlida>metformin>Maj or Bupleurum decoction.2.The results of network pharmacology showed that 43 active components of Jinqi Jiangtang Tablet corresponded to 81 targets of T2DM.The key active components of Jinqi Jiangtang Tablet for T2DM were magnoflorine,sinapic acid,luteolin,Dihydrochelerythrine and epiberberine.The PPI network construction and topology analysis of 81 targets showed that the core targets may be ALB,AKT1,VEGFA,SRC and EGFR.The results of GO enrichment analysis showed that Jinqi Jiangtang Tablet may play hypoglycemic effect through the regulation of protein kinase activity,nuclear receptor activity,transcription factor activity,steroid binding,etc.The enrichment analysis of KEGG pathway shows that Jinqi Jiangtang Tablet may regulate the resistance of EGFR tyrosine kinase inhibitor resistance,PI3K/Akt signaling pathway,Rapl signaling pathway,endocrine resistance signaling pathway,HIF-1 signaling pathway and other prevention and treatment of type 2 diabetes.Combined with the literature report and the correlation with type 2 diabetes,we selected PI3K Akt signaling pathway as the hypoglycemic pathway of Jinqi Jiangtang Tablet to improve insulin resistance in type 2 diabetes patients.3.The selected concentrations of 2 Jinqi Jiangtang tablets(400 ?g/mL?100 ?g/mL),compared with the model group,can increase their glucose Consumption(P<0.05).Western blot experiments and RT-PCR experiments found that the expression of key proteins,phosphorylation levels and mRNA expression of InsR-PI3K-Akt pathway in the model group were significantly down-regulated compared with the normal group(P<0.001).After 100 ?g/mL and 400 ?g/mL Jinqi Jiangtang tablets were applied to the insulin-resistant HepG2 cell model,the protein expression,phosphorylation level and mRNA expression level were significantly increased(P<0.05),indicating that Jinqi Jiangtang tablets improve cell glucose metabolism by enhancing the signal transduction of the InsR-PI3K-Akt pathway.ConclusionThe network meta-analysis results of Jinqi Jiangtang tablets showed that it had certain clinical efficacy.Through network pharmacology research,we predicted the pathway of Jinqi Jiangtang tablets' hypoglycemic effect,and selected InsR-PI3K-Akt signaling pathway as the target pathway for cell experiment verification,Cell model experiment showed that Jinqi Jiangtang Tablet could enhance the signal transduction of cells by enhancing the expression of key target proteins in InsR-PI3K-Akt signaling pathway,so as to improve the insulin resistance of model cells.This effect may be one of the mechanisms of its treatment of type 2 diabetes.