富含胱氨酸蛋白61对缺氧状态下肾小管上皮细胞的保护作用  

The protection effect of cysteine rich-protein 61 on renal tubular epithelial cells against apoptosis induced by hypoxia

作　　者：徐岩[1] 国敏[1] 董晖[1] 刘雪梅[1] 马瑞霞[1] 

机构地区：[1]青岛大学附属医院肾内科,青岛266000

出　　处：《中华肾脏病杂志》2015年第6期451-455,共5页Chinese Journal of Nephrology

基　　金：基金项目：国家自然科学基金面上项目（81170688、81470973）;山东省自然科学基金（ZR2011HM053）

摘　　要：目的 探讨富含胱氨酸蛋白61 （cysteine-rich protein 61,Cyr61）蛋白在缺氧缺血性急性肾损伤（AKI）中肾小管上皮细胞的表达,对肾小管上皮细胞的保护作用及可能机制.方法 以重组Cyr61慢病毒载体构建稳定表达Cyr61蛋白的人肾小管上皮细胞株Cyr61-HK2.溴脱氧尿苷核素掺入法检测细胞增殖,膜联蛋白V和碘化丙啶染色后流式细胞仪分析细胞凋亡,Western印迹检测BAD、Akt和ERK蛋白水平.结果 （1）与野生型HK2细胞相比,缺氧状态下Cyr61-HK2细胞的增殖能力更强;（2）缺氧状态下,野生型HK2细胞和Cyr61-HK2细胞的凋亡都随时间延长而增加,但与野生型HK2细胞相比,Cyr61-HK2细胞的凋亡减少;（3）与野生型HK2细胞相比,Cyr61-HK2在细胞缺氧0h、0.5 h、1h时的磷酸化BAD、磷酸化Akt和磷酸化ERK均增强,差异有统计学意义（均P＜0.05）.结论 缺氧状态下Cyr61蛋白能够促进肾小管上皮细胞增殖,抑制凋亡,Akt/ERK途径可能参与了Cyr61的抗凋亡作用.Objective To observe the expression of cysteine rich-protein 61 （Cyr61） on renal tubular cells,to explore its effects against hypoxic induced kidney injury and the underlying mechanisms.Methods A stably Cyr61 expressed tubular cell line Cyr61-HK2 was established based on HK2 cells and recombinant Cyr61-lentivirus.BrdU incorporation assay was used for cell proliferation.The apoptosis of cells was analyzed by flow cytometry with Annexin V and propidiumiodide staining.Western bloting was used to detect the protein expression of BAD,Akt and ERK.Results （1） Cyr61-HK2 cells displayed more proliferation ability than HK2 cells.（2） Under hypoxia condition,the apoptosis of both HK2 and Cyr61-HK2 cells increased,but the apoptosis of Cyr61-HK2 cells was lesser than HK2 cells.（3） The expression of Cyr61 led to the phosphorylation of BAD,Akt and ERK on 0 h,0.5 h,1 h （all P 〈 0.05）.Conclusion The expression of Cyr61 can promote cell proliferation and dampen cell apoptosis induced by hypoxia,which may be involved in the Akt/ERK signal pathway.

关 键 词：缺氧 细胞凋亡 急性肾损伤 富含胱氨酸蛋白61 

分 类 号：R692[医药卫生—泌尿科学]