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目的:探讨黄芪单体毛蕊异黄酮对转化生长因子β1(TGF-β1)诱导人脐静脉内皮细胞转分化的影响,以阐明黄芪毛蕊异黄酮对肾脏纤维化的保护作用。方法:人脐静脉内皮细胞系ECV-304为研究对象,体外培养ECV-304细胞株,分为正常对照组,TGF-β1组(10μg/L),TGF-β1+毛蕊异黄酮低、中、高剂量组(0.1、1、10mg/L)。各组细胞培养72h后,采用荧光倒置相差显微镜观察细胞形态学变化,Western Blot检测肌成纤维细胞标志物α平滑肌肌动蛋白(α-SMA)的表达变化。结果:与正常对照组相比,TGF-β1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,TGF-β1明显促进α-SMA蛋白的表达(P<0.01);毛蕊异黄酮与TGF-β1共培养后可明显抑制TGF-β1诱导的细胞形态学改变及α-SMA的表达(P<0.05),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:黄芪单体毛蕊异黄酮可以抑制TGF-β1诱导的内皮细胞(ECV-304)向肌成纤维细胞转化,具有保护内皮细胞的作用,从而为防治慢性肾脏病提供了依据。
OBJECTIVE: To investigate the effect of calrethin on the transdifferentiation of human umbilical vein endothelial cells induced by transforming growth factor-β1 (TGF-β1) in order to elucidate the protective effect of Astragalus curculia isoflavones on renal fibrosis. Methods: Human umbilical vein endothelial cell line ECV-304 was cultured in vitro and divided into normal control group, TGF-β1 group (10μg / L), TGF-β1 + Group (0.1, 1, 10 mg / L). The morphological changes of cells were observed by fluorescence inverted phase contrast microscope and the expression of α-smooth muscle actin (α-SMA), a marker of myofibroblast cells, was detected by Western Blot. Results: Compared with the normal control group, TGF-β1 cells transformed from the typical epithelial cell morphology to long spindle myofibroblast morphology. TGF-β1 significantly promoted the expression of α-SMA protein (P <0.01). Co-cultured with TGF-β1 could significantly inhibit the cell morphological changes and the expression of α-SMA induced by TGF-β1 (P <0.05). The high-dose group had stronger inhibitory effect than the low-dose and medium-dose groups The dose group had the weakest inhibitory effect. CONCLUSION: ACR can inhibit the transformation of endothelial cells (ECV-304) induced by TGF-β1 to myofibroblasts and protect the endothelial cells, which provides a basis for the prevention and treatment of chronic kidney disease.