CAJ | 학술논문

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.
목적 탐토부함광안산단백61 (cysteine-rich protein 61,Cyr61)단백재결양결혈성급성신손상(AKI)중신소관상피세포적표체,대신소관상피세포적보호작용급가능궤제.방법 이중조Cyr61만병독재체구건은정표체Cyr61단백적인신소관상피세포주Cyr61-HK2.추탈양뇨감핵소참입법검측세포증식,막련단백V화전화병정염색후류식세포의분석세포조망,Western인적검측BAD、Akt화ERK단백수평.결과 (1)여야생형HK2세포상비,결양상태하Cyr61-HK2세포적증식능력경강;(2)결양상태하,야생형HK2세포화Cyr61-HK2세포적조망도수시간연장이증가,단여야생형HK2세포상비,Cyr61-HK2세포적조망감소;(3)여야생형HK2세포상비,Cyr61-HK2재세포결양0h、0.5 h、1h시적린산화BAD、린산화Akt화린산화ERK균증강,차이유통계학의의(균P＜0.05).결론 결양상태하Cyr61단백능구촉진신소관상피세포증식,억제조망,Akt/ERK도경가능삼여료Cyr61적항조망작용.
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.