Objective:We aimed to observe the effects of loganin(Log)on serum glycolipid levels and probe the mechanisms focusing on intestinal flora and AMP-activated protein kinase(AMPK)signaling in obese mice.Methods:A high-fat diet was given for 12 consecutive weeks to generate the obesity model in institute of cancer research(ICR)mice.Body weight was measured weekly and fasting blood glucose(FBG)was determined every 2 weeks.Both the oral glucose tolerance test and the intraperitoneal insulin tolerance test were performed.The serum levels of total cholesterol(TC),triglyceride,high-density lipoprotein-cholesterol,low-density lipoprotein-cholesterol(LDL-C),and free fatty acids(FFA)were measured.The expression of key proteins in the AMPK signaling pathway in skeletal muscle tissue was detected by immunoblotting,and gut microbiota were characterized using 16S rDNA sequencing.Results:Log significantly decreased the body weight and the FBG in obese mice(P＜.05),and it could restore FBG to normal levels.The total cholesterol,LDL-C,and FFA levels were significantly reduced by Log compared with the obese controls(TC:P=.0020;LDL-C:P=.0233;FFA:P=.0127),and the glucose tolerance of animals was significantly improved(P=.0477).The western blot results showed that Log could upregulate the protein expression of Adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPKα),Sirtuin 1(SIRT1),and peroxisome proliferator-activated receptor-gamma coactivator-1 alpha(PGC1α)in skeletal muscle tissue of obese mice.16S rDNA sequencing indicated that Log reduced the diversity of the gut flora in feces and altered the floral composition of obese mice.Conclusions:Log was effective in reducing body weight and improving glucolipid metabolism in obese mice,probably through activating AMPK signaling and regulating intestinal microbial diversity.

OBJECTIVE: To predict the targets and pathways in the therapeutic mechanism of Guizhi Gancao Decoction (GZGCD) against heart failure (HF) based on network pharmacology. METHODS: The chemical components of GZGCD were analyzed using the databases including TCMSP, TCMID and TCM@Taiwan, and the potential targets of GZGCD were predicted using the SwissTargetPrediction database. The targets of HF were obtained using the databases including DisGeNET, Drugbank and TTD. The intersection targets of GZGCD and HF were identified using VENNY. Uniport database was used to convert the information, and the components-targets-disease network was constructed using Cytoscape software. The Bisogene plug-in, Merge plug-in, and CytoNCA plug-in in Cytoscape software were used for protein-protein interaction (PPI) analysis to acquire the core targets. Metascape database was used for GO and KEGG analysis. The results of network pharmacology analysis were verified with Western blot analysis. Three factors (PKCα, ERK1/2 and BCL2) were screened according to the degree value of network pharmacology results and the degree of correlation with heart failure process. The pentobarbtal sodium was dissolvein H9C2 cells treated with serum-free high glucose medium to simulate the ischemic anoxic environment of heart failure. The total proteins of myocardial cells were extracted. The protein contents of PKCα, ERK1/2 and BCL2 were determined. RESULTS: We identified a total of 190 intersection targets between GZGCD and HF using Venny database, involving mainly the circulatory system process, cellular response to nitrogen compounds, cation homeostasis, and regulation of the MAPK cascade. These potential targets were also involved in 38 pathways, including the regulatory pathways in cancer, calcium signal pathway, cGMP-PKG signal pathway, and cAMP signal pathway. Western blot analysis showed that in an in vitro H9C2 cell model of HF, treatment with GZGCD downregulated PKCα and ERK1/2 expressions and upregulated BCL2 expression. CONCLUSION The therapeutic mechanism of GZGCD for HF involves multiple targets including PRKCA, PRKCB, MAPK1, MAPK3, and MAPK8 and multiple pathways including the regulatory pathway in cancer and the calcium signaling pathway.
Humans ; Protein Kinase C-alpha ; Network Pharmacology ; Heart Failure/drug therapy* ; Proto-Oncogene Proteins c-bcl-2