松弛素对高糖诱导的肾小管上皮细胞转分化及分泌细胞外基质的影响

Effects of Relaxin on the Epithelial-to-mesenchymal Transition and the Production of ECM Excreted by High Glucose Stimulated Human Proximal Tubular Epithelial Cells

目的:观察人松弛素(Relaxin)对高糖培养人肾小管上皮细胞(HK-2)转分化、分泌细胞外基质的影响。方法:实验分组:正常糖组:含5.5 mmol/L葡萄糖;高糖组:含30 mmol/L葡萄糖;高渗组(甘露醇组);高糖+松弛素干预组;ELISA法检测细胞上清中TGF-β1、细胞胶原(ColⅠ)、纤连蛋白(FN)的表达。免疫印迹技术(Western blot)检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达,确定肾小管上皮细胞表型转化情况。结果:与正常糖组相比,高糖组肾小管上皮细胞分泌ColⅠ、FN、TGF-β1显著增加,在高糖培养基中加入人松弛素干预后可显著抑制由高糖诱导的ColⅠ、FN、TGF-β1的高表达,降低α-SMA表达,抑制肾小管上皮细胞转分化。结论:松弛素能抑制高糖诱导HK-2细胞分泌细胞外基质,抑制HK-2细胞转分化,具有抗纤维化作用。

Objective：To explore the effects of relaxin on the epithelial-to-mesenchymal transition and the production of ECM excreted by high glucose stimulated human proximal tubular epithelial cells.Methods：Cultured HK2 cells were divided into four groups：（1）normal glucose group（NG,5.5mmol/LD-glucose）,（2）manitol group（3）high glucose group（HG,30mmol/L D-glucose）,（4）High glucose＋relaxin group.The levels of fibronectin,collagen type Ⅰand TGF-β1 in the culture supernatants were examined by a solid-phase enzyme-linked immunoadsorbent assay（ELISA）;Western blot method was employed to detect the expression of α-smooth muscle actin（α-SMA） protein.Results：Compared with the normal glucose group,the levels of fibronectin,collagen typeⅠand TGF-β1 increased largely after cultured 48h in high concentration of glucose,while a remarkably decrease in the relaxin treated group.The expression ofα-smooth muscle actin was inhibited.Conclusion：Relaxin can suppress the overproduction of ECM excreted by HK-2 cultured in high ambient glucose,as well as restrain the procession of epithelial-myofibroblast transdifferentiation.