益智仁-乌药药对调控PI3K/Akt/mTOR通路介导细胞自噬保护肾小球足细胞的作用机制研究

Study on Mechanism of Yizhiren(Alpiniae Oxyphylae Fructus)-Wuyao(Linderae Radix)Drug Pair in Regulating PI3K/Akt/mTOR Pathway-Mediated Cellular Autophagy to Protect Podocytes

目的研究益智仁-乌药药对通过调控PI3K/Akt/mTOR信号通路促进足细胞自噬治疗糖尿病肾病(Diabetic Nephropathy,DN)的作用。方法60只造模成功的C57BL/KSJ-db/db(以下简称db/db)小鼠随机分为模型组、二甲双胍组、缬沙坦组、益智仁-乌药药对(低、中、高剂量)组,每组10只;另取10只C57BL/KSJ-db/m(以下简称db/m)小鼠为正常组,正常组和模型组给予生理盐水,治疗组小鼠分别给予相应药物,给药8周后检测小鼠肾脏病理学改变,足细胞自噬体数量、结构及相关蛋白表达。结果与模型组相比,益智仁-乌药药对组可显著减轻糖尿病肾病小鼠肾小球基底膜增厚情况,增加足细胞自噬体数量,显著升高自噬相关蛋白表达(P<0.05),降低PI3K/Akt/mTOR信号通路相关蛋白的表达(P<0.05)。其中益智仁-乌药药对高剂量组各指标改善优于益智仁-乌药低、中剂量组。结论益智仁-乌药药对通过抑制PI3K/Akt/mTOR信号通路激活,提高足细胞自噬水平,减轻足细胞损伤,发挥治疗糖尿病肾病的作用。

Objective To investigate the effect of Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)drug pair on regulating autophagy of podocyte through PI3K/Akt/mTOR signaling pathway in the treatment of diabetic nephropathy.Methods Sixty successfully modeled C57BL/KSJ-db/db(hereinafter referred to as db/db)mice were randomly divided into model group,metformin group,valsartan group and Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)drug pair(low,medium and high dose)groups,10 mice in each group.Another 10 C57BL/KSJ-db/m(hereinafter referred to as db/m)mice were taken as the normal group.The mice in the normal and model groups were given physiological saline,and the mice in the treatment groups were given the corresponding drugs,and the renal pathological changes,the number and structure of autophagosomes of podocyte and the related protein expression were examined after 8 weeks of administration.Results Compared with the model group,the Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)drug pair group significantly reduced glomerular basement membrane thickening,increased the number of autophagosomes in the podocyte,significantly increased the expression of autophagy-related proteins(P<0.05)and decreased the expression of PI3K/Akt/mTOR signaling pathway-related proteins(P<0.05).Among them,the improvement of all indexes in the Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)high dose group was better than that in the Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)low and medium dose groups.Conclusion The effect of Yizhiren(Alpiniae Oxyphyllae Fructus)-Wuyao(Linderae Radix)drug pair in the treatment of diabetic nephropathy is to inhibit the activation of PI3K/Akt/mTOR signaling pathway,increase the level of autophagy in podocyte and reduce podocyte damage in mice with diabetic nephropathy.