毛蕊异黄酮对高糖诱导内皮细胞转分化的影响及机制初探

Effect of Calycosin on High Glucose Induced Transdifferentiation of Endothelial Cells and its Mechanism

目的:探讨毛蕊异黄酮对高糖作用下人脐静脉内皮细胞转分化及转化生长因子β1(TGF-β1)分泌的影响,以阐明毛蕊异黄酮对糖尿病肾病的保护作用。方法:人脐静脉内皮细胞系(ECV-304)常规细胞培养传代,用于实验。细胞分为5组:高糖组:葡萄糖(GS)(4. 25%);毛瑞异黄酮组低剂量组:高糖(4. 25%GS)+毛蕊异黄酮(0. 1 mg/L);毛瑞异黄酮组中剂量组:高糖(4. 25%GS)+毛蕊异黄酮(1 mg/L);毛蕊异黄酮组高剂量组:高糖(4. 25%GS)+毛蕊异黄酮(10 mg/L);正常对照组:培养液中不加任何处理因素。各组加入干预因素后,在5%CO2和37℃条件下孵育48 h,分别收集细胞进行检测。荧光相差显微镜观察细胞形态学变化,Western印迹方法检测细胞α-SMA蛋白表达水平。ELISA法检测细胞培养上清液中TGF-β1的含量。结果:与正常对照组相比,高糖组细胞从原有典型的上皮细胞形态转变为长梭形肌成纤维细胞形态,高糖明显促进α-SMA蛋白的表达(P <0. 01)和TGF-β1分泌(P <0. 05);毛蕊异黄酮与高糖共培养后可明显抑制高糖诱导的细胞形态学改变及α-SMA表达(P <0. 05)和TGF-β1分泌(P <0. 05),并呈浓度依赖性。高剂量组比低、中剂量组有更强的抑制效应,低剂量组抑制效应最弱。结论:毛蕊异黄酮以浓度依赖性抑制内皮细胞转分化,这种效应可能与抑制TGF-β1分泌有关,从而为毛蕊异黄酮防治糖尿病肾病提供了依据。

Objective： This study aimed to investigate the protective effect of calycosin on the transdifferentiation of endothelial cells and transforming growth factor beta 1（TGF-β1） induced by high glucose,furthermore,to elucidate the protective mechanism of calycosin on diabetic kidney disease. Methods： A human umbilical endothelium derived celline（ECV-304） from CTCC was used in this study. Cultured endothelial cells were divided randomly into 5 groups ： control,high glucose（GS）（4. 25%）,（GS）（4. 25%） ＋ calycosin（0. 1,l,10 mg/L respectively）. Morphology of endothelial cells was studied by inverted fluorescence microscope. Expression of α-SMA was detected with Western Blot. TGF-β1 was detected with ELISA. Results： Compared with the control,cells morphology induced by high glucose changed from the original form of the typical epithelial cells into myofibroblasts spindle,high glucose significantly increased the expression of α-SMA（P〈0. 01） and TGF-β1（P〈0. 05）. Calycosin significantly inhibited cell morphological change and expression of α-SMA and TGF-β1（P〈0. 05） induced by high glucose in a dose-dependent manner. The high-dose calycosin group had a stronger inhibitory effect than the low and middle dose groups,the weakest inhibitory effect emerged in the low dose group. Conclusion： Calycosin showed a protective effect on high glucose induced transdifferentiation of endothelial cells,and this may provide beneficial action on the treatment of diabetic kidney disease.