激活素A及卵泡抑素对肾小管上皮细胞转分化的作用

目的探讨激活素A(ACTA)对大鼠肾小管上皮细胞(NRK-52E)转分化的影响及卵泡抑素(FS)的干预作用。方法 NRK-52E细胞生长于达氏修正伊氏培养基(DMEM)/F12培养基。实验A:将培养的NRK-52E细胞按rh-ACTA浓度不同分为4组:ACTA浓度分别为0,1,10,100ng/ml。实验B:分为4组:对照组;ACTA组(A组),rh-ACTA浓度10ng/ml;rh-FS组(F组),rh-FS浓度为100ng/ml;ACTA+FS组(A+F组):rh-ACTA浓度10ng/ml,rh-FS浓度为100ng/ml。24h后收集各组细胞及细胞上清液。蛋白印迹法检测各组细胞α-平滑肌肌动蛋白(α-SMA)和纤维连接蛋白(FN)的表达;酶联免疫吸附法(ELISA)检测各组细胞上清液转化生长因子-β(TGF-β)的含量。结果 ACTA能剂量依赖性刺激肾小管上皮细胞α-SMA及FN的表达(P<0.05),而对TGF-β的表达没有影响。FS能抑制ACTA的上述效应,而单纯FS对肾小管上皮细胞α-SMA及FN的表达无影响(P>0.05)。结论 ACTA能刺激NRK-52E细胞转分化及FN的合成;FS可通过阻断ACTA的上述效应产生有效的肾脏保护作用。

Effects of activin A and follistafin on renal tubular epithelial cells transdifferentiation

Objective To investigate the effects of activin A （ACT A） and follistatin （FS） on transdifferentiation of renal tubular epithelial cells （NRK-52E） in rats. Methods The NRK-52E cells were incubated in DMEM/F12 culture medium. In study A, NRK-52E cells were assigned to four groups according to the concentration of rh-ACT A （0, 1, 10, 100 ng/ml）. In study B, NRK-52E cells were incubated with negative control （group C）, 10 ng/ml rh-ACT A （group A）, 100 ng/ml rh-FS group （group F） and 10 ng/ml rh-ACT A plus 100 ng/ml rh-FS （group A＋F） for 24 hours. This entailed subsequent harvesting of the cells and cellular supernatant. Western blotting was applied to assay the a- smooth muscle actin （a-SMA） and fibronectin （FN） expression, and enzyme-linked immunosorbent assay （ELISA） was employed to quantitatively analyze the content of TGF-β Results ACT A stimulated expression of a-SMA and FN, but not TGF-β, in renal tubular epithelial cells in a dose-dependent fashion （P〈0.05）. Follistatin attenuated the over- expression of a-SMA and FN via inhibition of ACT A while did not alter the expression ofa-SMA and FN （P〉0.05）. Conclusion FS is characterized by the renal protective effects via blockade of NRK-52E ceils transdifferentiation and FN synthesis stimulated by ACT A.