桂枝通络片改善高脂诱导的ApoE-/-小鼠动脉粥样硬化伴骨质疏松的作用机制OA

Objective To investigate the mechanism of action of Guizhi Tongluo Tablets(GZTL)in ameliorating atherosclerosis(AS)with osteoporosis(OP)induced by a high-fat diet in ApoE-/-mice.Methods Mass spectrometry combined with the TCMSP database was used to predict the potential targets of GZTL.GO and KEGG enrichment analyses were performed using the DAVID database,and a PPI network was constructed to screen core targets.ApoE-/-mice were randomly divided into a model group,low-and high-dose GZTL groups,and an atorvastatin calcium(ATV)group.An AS combined with OP model was established by feeding a high-fat diet for 24 weeks.C57BL/6J mice served as the control group.Serum lipid levels were detected by biochemical methods.Histopathology was observed by HE staining.Femoral microstructure was examined by micro-CT.Serum levels of bone alkaline phosphatase(BALP)and osteocalcin were measured by ELISA.Protein and mRNA expression of PI3K/AKT pathway-related molecules were detected by Western Blot and qPCR.MC3T3-E1 cells were divided into a control group(osteogenic differentiation induction),a GZTL group(osteogenic differentiation induction+GZTL-containing serum),a LY294002 group(osteogenic differentiation induction+5 μmol·L-1 LY294002),and a GZTL+LY294002 group(osteogenic differentiation induction+GZTL-containing serum+5 μmol·L-1 LY294002).Cell proliferation rate was assessed by CCK-8 assay.Mineralized nodules were observed by Alizarin red staining.mRNA expression related to the PI3K/AKT signaling pathway was detected by qPCR.Results Bioinformatics analysis identified 408 potential target genes of GZTL for treating AS with OP,suggesting mechanisms involving lipid metabolism and the PI3K-AKT signaling pathway.In vivo experiments showed that compared with the model group,mice in the GZTL dose groups and the ATV group exhibited decreased serum lipid levels(P＜0.05,P＜0.01),improved lipid deposition and hepatic steatosis,significantly improved femoral microstructure(P＜0.05,P＜0.01),significantly increased serum BALP and osteocalcin levels(P＜0.05,P＜0.01),and significantly upregulated ratios of p-PI3K/PI3K,p-AKT/AKT,and mRNA expression of Pi3k and Akt in bone tissue(P＜0.05,P＜0.01).In vitro experiments showed that GZTL-containing serum promoted the proliferation and osteogenic differentiation of MC3T3-E1 cells,enhanced the formation of mineralized nodules,and significantly upregulated Pi3k and Akt mRNA levels(P＜0.01).These effects were partially antagonized by the PI3K inhibitor.Conclusion GZTL exerts a dual effect in the prevention and treatment of AS with OP by regulating lipid metabolism and promoting osteogenic differentiation.Its mechanism is closely related to the regulation of the PI3K/AKT signaling pathway.