柴胡皂苷D介导TGF-β/Smad信号通路对高糖诱导的HK-2细胞纤维化调节作用研究  

Regulatory effect of saikosaponin D-mediated TGF-β/Smad signaling pathway on high glucose induced HK-2 cell fibrosis

作　　者：胡采红 何泳 HU Caihong;HE Yong(Wuhan Hospital of China University of Geosciences,Wuhan 430070,China;Internal Medicine of Nephropathy,Wuhan Fifth Hospital,Wuhan 430050,China)

机构地区：[1]中国地质大学(武汉)医院,湖北武汉430070 [2]武汉市第五医院肾病内科,湖北武汉430050

出　　处：《沈阳药科大学学报》2022年第12期1471-1477,共7页Journal of Shenyang Pharmaceutical University

基　　金：武汉市卫生局科研项目(WX12C14)。

摘　　要：目的研究柴胡皂苷D对高糖诱导的人肾近曲小管上皮细胞HK-2的纤维化调节作用及其机制。方法将人肾近曲小管上皮细胞HK-2细胞分为对照组、模型组(27.5 mmol·L^(-1)的葡萄糖)、柴胡皂苷D组(27.5 mmol·L^(-1)的葡萄糖+1 mg·L^(-1)的柴胡皂苷D)和阳性对照组(27.5 mmol·L^(-1)的葡萄糖+0.2 g·L^(-1)吡非尼酮)。以免疫荧光的方法检测细胞中Ⅰ型胶原蛋白(collagen I,ColI)、纤连蛋白(fibronectin,FN)和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达。Western blot检测细胞中转化生长因子-1(transforming growth factor-1,TGF-β1)、Smad2、p-Smad2、Smad3和p-Smad3蛋白表达水平。结果和对照组比较,模型组的ColI、α-SMA、FN,TGF-β1、p-Smad2、p-Smad3蛋白表达水平均升高(P<0.05);和模型组比较,柴胡皂苷D组和阳性对照组细胞的ColI、α-SMA、FN、TGF-β1、p-Smad2、p-Smad3蛋白表达水平均显著下降(P<0.05)。结论柴胡皂苷D可缓解高糖诱导的HK-2细胞纤维化变化,其机制可能与抑制TGF-β1/Smad信号通路有关。Objective To study the regulatory effect of saikosaponin D on high glucose induced fibrosis in human renal proximal tubular epithelial cells HK-2 and its mechanism.Methods HK-2 cells were divided into control,model(27.5 mmol·L^(-1)glucose),saikosaponin D(27.5 mmol·L^(-1)glucose+1 mg·L^(-1)saikosaponin D),and positive control(27.5 mmol·L^(-1)glucose+0.2 g·L^(-1)pirfenidone)groups.The expression of type I collagen(ColI),fibronectin(FN)andα-smooth muscle actin(α-SMA)was detected by immunofluorescence.The protein expression levels of transforming growth factor-1(TGF-β1),Smad2,p-Smad2,Smad3 and p-smad3 were detected by Western blot.Results Compared with the control group,the expression levels of ColI,α-SMA,FN protein,TGF-β1,p-Smad2,p-smad3 protein in the model group were all increased(P<0.05).Compared with the model group,the expression levels of ColI,α-SMA,FN,TGF-β1,p-Smad2,p-smad3 protein in the saikosaponin D group and the positive control group were all decreased(P<0.05).Conclusion Saikosaponin D can alleviate the fibrosis of HK-2 cells induced by high glucose,and its mechanism may be related to the inhibition of TGF-β1/Smad signal pathway.

关 键 词：柴胡皂苷D 高糖 肾纤维化 TGF-Β/SMAD信号通路 

分 类 号：R917[医药卫生—药物分析学]