2022 年第 8 期 第 17 卷

阿司匹林对碘海醇诱导的肾损伤肾小管上皮细胞线粒体的保护作用及相关机制研究

Protective effect of aspirin on mitochondrion in renal tubular epithelial cells damaged by iohexol and its related mechanisms

作者：苟晓燕吕梁撒亚莲曾泽群

英文作者：Gou Xiaoyan Lyu Liang Sa Yalian Zeng Zequn

单位：昆明理工大学附属医院放射科，昆明650034

英文单位：Department of Radiology Affiliated Hospital of Kunming University of Science and Technology Kunming 650034 China

关键词：阿司匹林；碘海醇；人肾小管上皮细胞；线粒体

英文关键词：Aspirin;Iohexol;Humanrenaltubularepithelialcells;Mitochondrion

• 摘要：

• 目的 探索阿司匹林对碘海醇诱导的肾损伤肾小管上皮细胞线粒体的保护作用及相关机制。方法 将人肾小管上皮细胞（HK-2）分为正常对照组（Control组，HK-2细胞不特殊处理），碘海醇处理组（Iohexol组，HK-2细胞+18.86 mgI/ml的碘海醇），阿司匹林组处理组（aspirin组，HK-2细胞+2.5 μmol/L的阿司匹林），碘海醇+阿司匹林处理组（Iohexol2组，HK-2细胞+18.86 mgI/ml的碘海醇+2.5 μmol/L的阿司匹林）。电镜观察各组细胞线粒体融合、分裂情况；流式细胞术检测细胞凋亡；免疫荧光检测线粒体活性氧自由基（ROS）的表达；收集HK-2细胞进行真核有参转录组测序分析；蛋白印迹法验证通路相关蛋白表达。结果 与Control组比较，Iohexol组和aspirin组中线粒体分裂增多、融合减少；与Iohexol组比较，Iohexol2组及aspirin组线粒体分裂减少、融合增多。Iohexol2组HK-2细胞凋亡率低于Iohexol组和aspirin组［（26.5±1.3）%比（71.8±4.3）%、（32.3±5.0）%］(P＜0.001、P=0.041)。Iohexol2组中ROS生成少于Iohexol组(P=0.002)。真核有参转录组测序分析结果显示，Iohexol组与Iohexol2组细胞丝裂原活化蛋白激酶通路具有明显差异。Iohexol2组细胞外调节蛋白激酶1/2（ERK1/2）与磷酸化ERK1/2比值、c-MYC结合蛋白及线粒体分裂动力相关蛋白1(DRP1)等蛋白表达水平均低于Iohexol组及aspirin组（均P＜0.01）。结论 阿司匹林可通过介导ERK1/2/c-MYC/DRP1信号通路减少碘海醇诱导的HK-2细胞线粒体的分裂。

• Objective To explore the protective effect of aspirin on mitochondrion in renal tubular epithelial cells damaged by iohexol and its related mechanisms. Methods Human renal tubular epithelial cells (HK-2) were divided into normal control group (control group, HK-2 cells without special treatment), iohexol treatment group (Iohexol group, HK-2 cells + 18.86 mgI/ml iohexol), aspirin treatment group (aspirin group, HK-2 cells+2.5 μmol/L aspirin), iohexol+aspirin treatment group (Iohexol2 group, HK-2 cells+18.86 mgI/ml iohexol+2.5 μmol/L aspirin). Mitochondrial fusion and division after experimental treatment were observed by transmission electron microscope. Cell apoptosis was detected by flow cytometry. Mitochondrial reactive oxygen species (ROS) in HK-2 cells was detected by immunofluorescence. HK-2 cells were collected for transcriptome gene sequencing analysis. The expression of pathway-related proteins was verified by western blotting. Results Compared with control group, mitochondrial division increased and fusion decreased in Iohexol group and aspirin group. Compared with Iohexol group, mitochondrial division decreased and fusion increased in Iohexol2 group and aspirin group. The apoptosis rate of HK-2 cells in Iohexol2 group was lower than that in Iohexol group and aspirin group ［(26.5±1.3)% vs (71.8±4.3)%, (32.3±5.0)%］(P＜0.001, P=0.041). The production of ROS in Iohexol2 group was less than that in Iohexol group(P=0.002). Transcriptome gene sequencing analysis showed that there were significant differences in mitogen activated protein kinase pathway between Iohexol group and Iohexol2 group. The expression levels of extracellular regulated protein kinase 1/2 (ERK1/2) to phosphorylated ERK1/2 ratio, c-MYC binding protein and mitochondrial division dynamics related protein 1 (DRP1) in Iohexol2 group were significantly lower than those in Iohexol group and aspirin group (all P＜0.01). Conclusion Aspirin can reduce mitochondrial division induced by iohexol in HK-2 cells by mediating ERK1/2/c-MYc/DRP1 signaling pathway.