首页|远隔缺血预处理分离液对人肾小管上皮细胞缺氧性损伤的保护作用

远隔缺血预处理分离液对人肾小管上皮细胞缺氧性损伤的保护作用

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目的:研究远隔缺血预处理(RIPC)分离液对人肾小管上皮细胞(HK-2)缺氧/复氧(H/R)损伤的影响及机制.方法:将HK-2 细胞分为时间对照组、H/R、RIPC处理组、自噬干预组.时间对照组细胞在正常条件下培养,RIPC组细胞使用RIPC分离液预处理,自噬干预组细胞在RIPC处理基础上,加入自噬抑制剂 3-甲基腺嘌呤(3-methyladenine,3MA),后三组细胞使用三气培养箱进行H/R构建损伤模型.模型成功后使用光学显微镜观察各组细胞状态,应用Western blot观察LC3Ⅱ/Ⅰ蛋白、Beclin1 蛋白的表达水平.结果:镜下示RIPC分离液可明显减轻H/R对HK-2 细胞的损伤,加入自噬抑制剂,RIPC分离液对HK-2 细胞H/R损伤的保护作用被阻断.Western blot结果显示,RIPC 分离液可使 HK-2 细胞线粒体中的自噬特异性蛋白LC3Ⅱ/Ⅰ、Beclin1 表达水平增高.Real-time PCR结果显示,与H/R组比,RIPC组LC3 及Bec-lin1 的mRNA表达量明显升高;加入自噬抑制剂,HK-2 细胞线粒体中的自噬特异性蛋白LC3Ⅱ/Ⅰ、Beclin1 表达降低,LC3 及Beclin1 的mRNA表达量同时也明显降低.结论:RIPC分离液对HK-2 细胞H/R后损伤具有保护作用,作用机制可能与线粒体自噬相关.
Protective effect of remote ischemic preconditioning separating medium on hypoxic injury of human renal tubular epithelial cells
Objective:To investigate the effect and mechanism of remote ischemic preconditioning(RIPC)separa-ting medium on hypoxia/reoxygenation(H/R)injury of human renal tubular epithelial cells(HK-2).Methods:HK-2 cells were divided into time control group,H/R,RIPC treatment group and autophagy intervention group.Cells in the time control group were cultured under normal conditions.Cells in the RIPC group were pretreated with RIPC separa-ting medium.Cells in the autophagy intervention group were treated with autophagy inhibitor 3-methyladenine(3MA)on the basis of RIPC treatment.Cells in the latter three groups were subjected to H/R in a three-gas incubator to con-struct a damage model.After the success of the model,the cell status of each group was observed by optical microscope,and the expression levels of LC3Ⅱ/Ⅰprotein and Beclin1 protein were observed by Western blot.Results:Under the microscope,RIPC separating medium could significantly reduce the damage of H/R to HK-2 cells.After adding auto-phagy inhibitor,the protective effect of RIPC separating medium on H/R injury of HK-2 cells was blocked.The results of Western blot showed that RIPC separating medium could increase the expression levels of autophagy-specific proteins LC3Ⅱ/Ⅰand Beclin1 in mitochondria of HK-2 cells.The results of Real-time PCR showed that the mRNA expres-sion of LC3 and Beclin1 in RIPC group was significantly higher than that in H/R group;the expression of autophagy-specific proteins LC3Ⅱ/Ⅰand Beclin1 in mitochondria of HK-2 cells was decreased by adding autophagy inhibitors,and the mRNA expression of LC3 and Beclin1 was also significantly decreased.Conclusion:RIPC separating medium has a protective effect on HK-2 cells after H/R injury,and the mechanism may be related to mitophagy.

Remote ischemic preconditioning separating mediumHuman renal tubular epithelial cellsHypoxia/reoxygenationMitochondrial autophagy

戴文俊、裴汉军

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内蒙古科技大学包头医学院,内蒙古 包头 014040

远隔缺血预处理分离液 人肾小管上皮细胞 缺氧/复氧 线粒体自噬

国家自然科学基金

81760034

2024

包头医学院学报
内蒙古科技大学包头医学院

包头医学院学报

影响因子:0.543
ISSN:1006-740X
年,卷(期):2024.40(2)
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