摘要
目的:探讨脂氧素A4(lipoxin A4,LXA4)在人肾脏近曲小管上皮细胞(HK-2)缺氧/复氧损伤中的保护作用及可能机制。方法:LXA4预处理HK-2细胞后进行缺氧/复氧处理,CCK-8法检测细胞活性水平,ELISA法检测细胞上清液γ-谷氨酰转肽酶(γ-GT)、氨基葡萄糖苷酶(NAG)和亮氨酸氨基肽酶(LAP)水平,羟胺法检测细胞超氧化物歧化酶(SOD)活性,硫代巴比妥酸法检测细胞丙二醛(MDA)水平,实时定量PCR和Western blot法分别检测HK-2细胞的过氧化物酶体增殖子活化受体γ(PPARγ)和血红素加氧酶-1(HO-1)的m RNA和蛋白表达的变化,利用RNA干扰技术干扰PPARγ表达后检测HO-1和核因子E2相关因子2(Nrf2)的表达。结果:LXA4预处理的缺氧/复氧组细胞活性和SOD活性明显升高,细胞中γ-GT、NAG、LAP和MDA水平降低,而加入HO-1抑制剂锌原卟啉(Zn PP)和转染PPARγ的si RNA后,LXA4对HK-2细胞缺氧/复氧损伤的保护作用被阻断;此外,LXA4预处理的缺氧/复氧组HO-1、PPARγ和Nrf2表达均明显升高,并且LXA4预处理诱导的HO-1和Nrf2过表达能够被PPARγsi RNA所抑制。结论 :Lx A4预处理能够通过诱导HK-2细胞的HO-1高表达对抗HK-2细胞缺氧/复氧损伤,其机制与激活PPARγ/Nrf2有关。
Objective:To investigate the effects and mechanisms of lipoxin A4(LXA4)in attenuating hypoxia / reoxygenation injury in human renal tubular epithelial cells(HK-2). Methods:HK-2 cells were exposed to hypoxia followed by reoxygenation with pretreatment of LXA4. Then the cell viability,γ-GT,NAG and LAP levels,SOD activity and MDA level were determined. The expressions of m RNA and protein of PPARγ and HO-1 were measured using real-time polymerase chain reaction(PCR) and Western blot,respectively. The expressions of HO-1 and Nrf2 were detected by using RNA interference technology through interference PPARγ expression. Results:Pretreatment with LXA4 increased the cell viability and SOD activity,and reduced the γ-GT,NAG,LAP and MDA levels. HO-1 inhibition by Zn PP and si RNA of PPARγ abolished the protective role of LXA4 on the cells undergoing hypoxia / reoxygenation injury. Furthermore,the expressions of HO-1,PPARγ and Nrf2 were increased in the cells pretreated with LXA4 significantly,whereas these overexpressions of HO-1 and Nrf2 were partly blocked by treatment with si RNA of PPARγ.Conclusion:This study reveals that LXA4 pretreatment serves a protective role against hypoxia / reoxygenation injury of human renal tubular epithelial cells via over-expression of HO-1 and activation of PPARγ / Nrf2.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2015年第8期1080-1086,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助(81270821)
