论文部分内容阅读
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 protective effect and possible mechanism of lipoxin A4 (LXA4) on hypoxia / reoxygenation injury in human renal proximal tubule epithelial cells (HK-2). METHODS: HK-2 cells were pretreated with LXA4 for hypoxia / reoxygenation. Cell viability was determined by CCK-8 assay. The levels of γ-glutamyl transpeptidase (γ-GT) and glucosamine (NAG) and leucine aminopeptidase (LAP), the activity of superoxide dismutase (SOD) was detected by hydroxylamine method, the level of malondialdehyde (MDA) was detected by thiobarbituric acid method, PCR and Western blot were used to detect the mRNA and protein expression of peroxisome proliferator-activated receptor γ (PPARγ) and heme oxygenase-1 (HO-1) in HK-2 cells respectively. The expression of HO-1 and NF-kappa2 factor 2 (Nrf2) was detected by interfering with the expression of PPARγ. RESULTS: The cell viability and SOD activity in LXA4 preconditioning hypoxia / reoxygenation group were significantly increased, while the levels of γ-GT, NAG, LAP and MDA were decreased in the cells pretreated with LXA4, while the addition of HO-1 inhibitor zinc protoporphyrin (Zn PP) LXA4 could inhibit the hypoxia / reoxygenation injury of HK-2 cells, and the expression of HO-1, PPARγ and Nrf2 in LXA4-treated hypoxia / reoxygenation group were significantly increased And LXA4 pretreatment induced HO-1 and Nrf2 overexpression can be inhibited by PPARγsi RNA. CONCLUSION: Lx A4 pretreatment can antagonize the hypoxia / reoxygenation injury in HK-2 cells by inducing HO-1 overexpression in HK-2 cells. The mechanism is related to the activation of PPARγ / Nrf2.