基于CaSR表达及AKT磷酸化水平的花旗泽仁改善胰岛素抵抗作用机制

目的 基于CaSR表达及AKT磷酸化水平考察花旗泽仁改善胰岛素抵抗的作用机制.方法 采用高浓度胰岛素体外诱导培养法建立IRL02肝细胞模型,并通过葡萄糖消耗实验对细胞模型进行鉴定.采用免疫荧光法和qRT-PCR技术检测CaSR表达的细胞定位以及CaSR mRNA表达,同时采用Western blot技术检测CaSR蛋白...

Action mechanism of Huaqizeren in improving insulin resistance based on CaSR expression and AKT phosphorylated level

AIM To investigate the action mechanism of Huaqizeren in improving insulin resistance based on CaSR expression and AKT phosphorylated level.METHODS The insulin resistance cell models of L02 hepatocytes induced by high-concentration insulin in vitro culture was identified by a glucose consumption experiment.Immunofluorescence and qRT-PCR were used to detect the localization of CaSR expression and CaSR mRNA expression.Meanwhile,Western blot was used to detect CaSR protein and phosphorylated AKT(Ser473 and Thr308)protein expression levels.RESULTS The insulin resistance L02 hepatocyte models displayed significantly decreased glucose consumption and uptake rate(P<0.01).The results of immunofluorescence experiments showed relatively sparse CaSR distribution in the model control group,but more dense distribution in the group intervened with Huaqizeren.The qRT-PCR experiments revealed significantly decreased CaSR mRNA expression levels in the model control group(P<0.01);but significantly increased CaSR mRNA expression levels in both Huaqizeren group and the positive control group in contrast to the model control group(P<0.01).Simultaneously,Western blot results showed significantly reduced CaSR and phosphorylated AKT(Thr308 and Ser473)protein expressions in the model control group(P<0.01),but significantly increased such protein expressions in both Huaqizeren group and the positive group when in comparison with the model group(P<0.01,P<0.05).CONCLUSION Huaqizeren’s efficacy confirmed in the experiment in insulin resistance improvement through its influence on the PI3K pathway and CaSR activation,associates with the protein expression of two phosphorylation sites(Thr308,Ser473)on the AKT of the PI3K/AKT signaling pathway.