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目的探讨核心蛋白聚糖对转化生长因子β_1(TGF-β_1)诱导的人肾小管上皮细胞(HK-2)转分化的影响。方法将体外培养的 HK-2细胞分为6组:阴性对照组(A 组);10 ng/mlTGF-β_1组(B 组);10 ng/ml 核心蛋白聚糖组(C 组);100 ng/ml 核心蛋白聚糖组(D 组);10 ng/mlTGF-β_1+10 ng/ml 核心蛋白聚糖组(E 组);10 ng/ml TGF-β_1+100 ng/ml 核心蛋白聚糖组(F 组)。倒置显微镜下观察加入刺激因子48 h 后,各组细胞形态学变化;应用逆转录-聚合酶链反应(RT-PCR)检测核心蛋白聚糖对 TGF-β_1诱导的 HK-2细胞内α平滑肌肌动蛋白(α-SMA)、波形蛋白、角蛋白mRNA 表达的影响。结果 B 组与 A 组相比较,细胞形态发生明显变化,大部分细胞拉长呈梭形,似成纤维细胞;E 组和 F 组比 B 组梭形样细胞明显减少,尤其是 F 组梭形样细胞减少更明显;C 组和 D组与 A 组细胞形态无明显区别,为椭圆形;B 组与 A 组比较,波形蛋白、α-SMA mRNA 表达明显提高,角蛋白 mRNA 表达减少。B 组平均值分别为(0.828±0.297)、(0.332±0.025)、(0.075±0.040),A组平均值分别为(0.033±0.060)、(0.003±0.000)、(0.348±0.012),差异有统计学意义(P<0.05);E 组和 F 组与 B 组相比,波形蛋白、α-SMA 的表达有所下降,角蛋白的表达呈上升趋势(P<0.05),平均值分别为 E 组(0.234±0.313)、(0.214±0.196)、(0.098±0.056),F 组(0.155±0.053)、(0.122±0.130)、(0.215±0.122)。C 组和 D 组与 A 组比较差异无统计学意义(P>0.05)。结论 TGF-β_1能诱导正常人肾小管上皮细胞发生转分化;核心蛋白聚糖对 TGF-β_1诱导的 HK-2细胞转分化具有抑制作用,且呈剂量依赖性。
Objective To investigate the effect of decorin on the transdifferentiation of human renal tubular epithelial cells (HK-2) induced by transforming growth factor-β1 (TGF-β1). Methods HK-2 cells cultured in vitro were divided into 6 groups: negative control group (group A), 10 ng / ml TGF-β 1 group (group B), 10 ng / ml decorin group 10 ng / ml TGF-β 1 + 10 ng / ml decorin group (E group); 10 ng / ml TGF-β 1 + 100 ng / ml decorin group (Group F). The morphological changes of cells in each group were observed under an inverted microscope 48 hours after addition of stimulus factors. Expression of decorin induced by TGF-β 1 in α-smooth muscle cells of HK-2 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) Effects of α-SMA, vimentin, and keratin mRNA expression. Results Compared with group A, the morphology of cells in group B changed significantly, most of the cells elongated in a fusiform shape and fibroblasts; the number of fusiform cells in group E and F decreased significantly compared with group B, especially in group F Like cells decreased more obviously. There was no significant difference between group C and group D and group A, which was oval. Compared with group A, the expression of vimentin and α-SMA mRNA was significantly increased and the expression of keratin mRNA was decreased. The average values of group B were (0.828 ± 0.297) and (0.332 ± 0.025) and (0.075 ± 0.040) respectively, and those of group A were (0.033 ± 0.060) and (0.003 ± 0.000) and (0.348 ± 0.012) (P <0.05). Compared with group B, the expression of vimentin and α-SMA decreased in E group and F group, and the expression of keratin increased (P <0.05), and the average values were E Group (0.234 ± 0.313), (0.214 ± 0.196), (0.098 ± 0.056), F group (0.155 ± 0.053), (0.122 ± 0.130) and (0.215 ± 0.122). There was no significant difference between group C and group D and group A (P> 0.05). Conclusion TGF-β 1 can induce normal human renal tubular epithelial cells to undergo transdifferentiation. Decorin has an inhibitory effect on the transdifferentiation of HK-2 cells induced by TGF-β 1 in a dose-dependent manner.