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目的:探讨丹酚酸B对转化生长因子(TGF)-β1诱导的肾小管上皮细胞—间充质细胞转分化的影响。方法:将体外培养的人近端肾小管上皮细胞系(HK-2)细胞分为3组:①对照组:未加入丹酚酸B或者TGF-β1;②TGF-β1组:在细胞培养基中加入TGF-β1(浓度为5ng/ml);③丹酚酸B+TGF-β1组:在细胞培养基中分别加入丹酚酸B和TGF-β1(浓度为5ng/ml),按丹酚酸B的浓度分为A、B、C、D4个亚组:丹酚酸B的浓度分别为A组:0.1μmol/L,B组:1μmol/L,C组:10μmol/L,D组:100μmol/L。72h后,采用倒置相差显微镜观察细胞形态学变化;采用逆转录聚合酶链反应(RT-PCR)和免疫细胞化学染色检测细胞α-平滑肌肌动蛋白(α-SMA)和E-cadherin的表达情况。结果:TGF-β1组HK-2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;胞浆内大量表达α-SMA,同时E-cadherind的表达明显减少;不同剂量丹酚酸B治疗可减轻TGF-β1诱导的细胞形态学改变和胞浆内α-SMA的表达,同时E-cadherind的表达得到明显恢复,且呈剂量依赖性。结论:丹酚酸B具有阻止慢性肾脏疾病进行性发展的潜能,而这一作用与其能有效阻止TGF-β1诱导的HK-2细胞转分化有关。
Objective: To investigate the effect of salvianolic acid B on the transdifferentiation of renal tubular epithelial-mesenchymal cells induced by transforming growth factor (TGF) -β1. Methods: Human proximal tubular epithelial cell line (HK-2) cells cultured in vitro were divided into three groups: ① control group: no salvianolic acid B or TGF-β1; ②TGF-β1 group: in the cell culture medium The cells were treated with TGF-β1 (5ng / ml); ③the salvianolic acid B + TGF-β1 group: salvianolic acid B and TGF-β1 The concentrations of B were divided into A, B, C and D subgroups. The concentrations of salvianolic acid B were 0.1 μmol / L in group A, 1 μmol / L in group B, 10 μmol / L in group C, / L. 72h later, the morphological changes of cells were observed by inverted phase contrast microscope. The expression of α-smooth muscle actin (α-SMA) and E-cadherin were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry . Results: HK-2 cells in TGF-β1 group changed from the original epithelial cells to long spindle myofibroblasts. Large amount of α-SMA was expressed in the cytoplasm and the expression of E-cadherind was significantly reduced. Treatment with phenolic acid B reduced the morphological changes of cells induced by TGF-β1 and the expression of α-SMA in cytoplasm. At the same time, the expression of E-cadherin was significantly restored in a dose-dependent manner. CONCLUSION: Salvianolic acid B has the potential to prevent the progressive development of chronic kidney disease, which is related to its ability to prevent TGF-β1-induced transdifferentiation of HK-2 cells.