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目的:通过观察花旗泽仁对2型糖尿病胰岛素抵抗大鼠肌肉组织中钙敏感受体(CaSR)基因、蛋白表达及AKT活性的影响,探究花旗泽仁治疗胰岛素抵抗的作用机制。方法:取雄性Wistar大鼠持续灌胃脂肪乳、腹腔注射链脲佐菌素的方式复制2型糖尿病胰岛素抵抗模型。将造模成功的胰岛素抵抗大鼠随机分为模型对照组、阳性对照组、花旗泽仁组。花旗泽仁组给予花旗泽仁水煎液,阳性对照组给予罗格列酮药液。给药结束后,分别检测各组大鼠空腹血糖(FBG)及空腹血清胰岛素(FINS),计算胰岛素敏感指数(ISI),并且取大鼠肌肉组织,采用qRT-PCR和Western blot技术检测钙敏感受体CaSR mRNA、CaSR蛋白和磷酸化AKT(Ser473和Thr308)蛋白表达水平。结果:胰岛素抵抗大鼠模型被成功建立。CaSR mRNA、CaSR蛋白表达水平明显降低(P<0.001),磷酸化AKT(Ser473和Thr308)蛋白含量明显下降(P<0.001,P<0.01);花旗泽仁组与模型组比较,FBG及FINS水平明显降低(P<0.01),ISI显著升高(P<0.01),CaSR mRNA、CaSR蛋白和磷酸化AKT(Ser473和Thr308)蛋白表达水平均明显增加(P<0.01,P<0.05)。结论:花旗泽仁可能通过上调CaSR表达水平,上调与胰岛素抵抗关系密切的PI3K/AKT信号通路中的AKT蛋白(Ser473、Thr308位点)磷酸化水平,从而改善胰岛素抵抗。
Objective: To observe the effect of Citizen’s Zerelin on the gene and protein expression of calcium-sensitive receptor (CaSR) and the activity of AKT in muscular tissue of type 2 diabetic rats with insulin resistance and to explore the mechanism of Citizen’s treatment of insulin resistance. Methods: Male Wistar rats were intragastrically administered with fat emulsion and streptozotocin intraperitoneally to replicate the insulin resistance model of type 2 diabetes mellitus. The rats with successful insulin resistance were randomly divided into model control group, positive control group and Citizen Zeren group. Citizen Zeren given Citrus Ze Yan decoction, the positive control group was given rosiglitazone solution. After the administration, fasting blood glucose (FBG) and fasting serum insulin (FINS) of rats in each group were measured, and insulin sensitivity index (ISI) was calculated. Muscle tissues of rats were taken out and calcium sensitivity was detected by qRT-PCR and Western blot Receptor CaSR mRNA, CaSR protein and phosphorylated AKT (Ser473 and Thr308) protein expression levels. Results: The rat model of insulin resistance was successfully established. The levels of CaSR mRNA and CaSR protein were significantly decreased (P <0.001) and the protein levels of phosphorylated AKT (Ser473 and Thr308) were significantly decreased (P <0.001, P <0.01). Compared with model group, the levels of FBG and FINS (P <0.01). The ISI was significantly increased (P <0.01). The protein expression of CaSR mRNA, CaSR protein and phosphorylated AKT (Ser473 and Thr308) were significantly increased (P <0.01, P <0.05). CONCLUSION: Citi Zeiren may improve insulin resistance by up-regulating CaSR expression and up-regulating phosphorylation of AKT protein (Ser473, Thr308) in PI3K / AKT signaling pathway closely related to insulin resistance.