MSN对血管紧张素Ⅱ诱导的HEK293细胞ColⅠ表达的影响

Effect of MSN on angiotensin Ⅱ-induced collagen I expression in HEK293 cells

目的:探讨MSN对血管紧张素Ⅱ诱导的人胚肾成纤维细胞(HEK293)ColⅠ表达的影响。方法:将浓度为1×108/L的HEK293细胞悬液接种1mL在6孔板中1cm×1cm的打胶盖玻片上,用透射电镜观察细胞超微结构,同步化细胞后分为正常对照组,阳性对照组,MSN高、中、低剂量组和溶液对照组,各组加入相应药物干预24h,免疫组化法检测ColⅠ表达,将染色后的细胞爬片应用病理图集采集和分析系统软件半定量分析结果。结果:ColⅠ在正常人胚肾成纤维细胞包浆呈弱阳性含量,在阳性对照组AngⅡ刺激下呈强阳性含量;MSN高、中、低剂量组与阳性对照组相比,胞浆颜色均变淡,表达变弱,MSN低剂量组与溶液对照组无明显差异。ColⅠ光密度值阳性对照组高于正常对照组(P<0.05);MSN各剂量组均低于阳性对照组(P<0.05)。结论:MSN通过抑制ColⅠ的含量,从而抑制系膜基质增生,抑制慢性肾衰的纤维化发生和进展。

Objective:To investigate the effect of MSN on angiotensin II(Ang II)-induced collagen I(Col I) expression in human embryonic kidney 293(HEK293) cells.Methods:HEK293 cell suspension(1 × 108 / L,1 ml) was inoculated onto coated cover slips(1cm ×1cm) in 6-well plates,and the cellular ultrastructure was observed under a transmission electron microscope.After being synchronized,HEK293 cells were divided into normal control group,positive control group,high-,middle-,and low-dose MSN groups,and solution control group.All groups received respective medical interventions for 24 h.The Col I expression was measured by immunohistochemistry.The stained cells on cover slips were semi-quantitatively analyzed by the pathological image collection and analysis software system.Results:Normal HEK293 cells were weakly positive for Col I,which was expressed in the cytoplasm,while the cells in positive control group,which were stimulated by Ang II,were strongly positive for Col I.Compared with the positive control group,the high-,middle-,and low-dose MSN groups had lighter cytoplasm and weaker expression of Col I;there was no significant difference between the low-dose MSN group and solution control group.The positive control group had a significantly higher optical density of Col I than the normal control group(P < 0.05);the high-,middle-,and low-dose MSN groups had significantly lower optical densities of Col I than the positive control group(P < 0.05).Conclusion:MSN inhibits Col I expression and thus mesangial matrix proliferation,so it can delay the development and progression of fibrosis in chronic renal failure.