咪达唑仑对低氧/复氧诱导的HK-2细胞凋亡及Nrf2/HO-1通路的影响

Effects of Midazolam on the Apoptosis and Nrf2/HO-1 Pathway in HK-2 Cells Induced by Hypoxia/Reoxygenation

目的:探讨咪达唑仑对低氧/复氧诱导HK-2细胞凋亡的作用,及对核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)通路的影响。方法:HK-2细胞分为正常对照组、低氧/复氧组、不同浓度(2,4,8μmol·L;)咪达唑仑组,CCK-8法检测细胞活性,流式细胞术检测细胞凋亡情况,试剂盒检测细胞活性氧簇(ROS)和丙二醛(MDA)含量以及超氧化物歧化酶(SOD)活性,蛋白质免疫印迹技术(WB)检测HK-2细胞Nrf2和HO-1表达水平。结果:相较于正常对照组,低氧/复氧组细胞活性及SOD活性明显降低(P<0.05),细胞凋亡率、ROS和MDA含量明显升高(P<0.05)。相较于低氧/复氧组,咪达唑仑各浓度组细胞活性、SOD活性及Nrf2和HO-1表达明显升高(P<0.05),细胞凋亡率、ROS和MDA含量明显降低(P<0.05),且呈剂量依赖性。结论:咪达唑仑可促进Nrf2/HO-1信号通路,提高细胞抗氧化能力,抑制低氧/复氧诱导的HK-2细胞凋亡。

Objective: To investigate the effects of midazolam on the apoptosis and Nrf2/HO-1 pathway in HK-2 cells induced by hypoxia/reoxygenation. Methods: HK-2 cells were divided into normal control group, hypoxia/reoxygenation group and midazolam groups with different concentrations(2, 4, 8 μmol·L;). CCK-8 assay and flow cytometry were used to detect cell viability and cell apoptosis, respectively, the contents of reactive oxygen species(ROS) and malondialdehyde(MDA), and the activity of superoxide dismutase(SOD) were detected by the kits, and the expression levels of NF-E2 related factor(Nrf2) and heme oxygenase-1(HO-1) in HK-2 cells were detected by Western blot(WB). Results: Compared with those in the normal control group, the cell activity and SOD activity of hypoxia/reoxygenation group were significantly decreased(P<0.05), and the apoptosis rate, contents of ROS and MDA were significantly increased(P<0.05);compared with those in hypoxia/reoxygenation group, the cell activity, SOD activity and Nrf2, HO-1 expression of midazolam groups were significantly increased(P<0.05), and the apoptosis rate, contents of ROS and MDA were significantly decreased(P<0.05). The effects of midazolam in each group were dose-dependent. Conclusion: Midazolam can promote Nrf2/HO-1 signaling pathway, enhance cell antioxidant capacity and inhibit the apoptosis of HK-2 cells induced by hypoxia/reoxygenation.