摘要
目的:研究沉默信息调节因子1(SIRT1)对缺氧复氧肾小管上皮细胞炎性损伤的影响。方法:构建缺氧复氧肾小管上皮细胞损伤模型,用qRT-PCR和Western blot方法检测细胞中SIRT1的表达变化。在肾小管上皮细胞中转染pc DNA3. 1-SIRT1,qRT-PCR和Western blot检测过表达效率。ELISA法检测细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平。Western blot检测CCAAT/增强子结合蛋白同源蛋白(CHOP)、Caspase-11蛋白水平。全自动生化分析仪检测培养液中乳酸脱氢酶(LDH)含量。结果:缺氧复氧肾小管上皮细胞中SIRT1 mRNA和蛋白水平均降低。pc DNA3. 1-SIRT1转染后的肾小管上皮细胞中SIRT1 mRNA和蛋白水平均明显升高。缺氧复氧肾小管上皮细胞分泌的TNF-α、IL-1β水平升高,细胞中CHOP、Caspase-11蛋白水平升高,细胞培养液中LDH水平升高。过表达SIRT1的肾小管上皮细胞经缺氧复氧处理以后,细胞分泌的TNF-α、IL-1β水平降低,细胞中CHOP、Caspase-11蛋白水平降低,细胞培养液中LDH水平降低。结论:SIRT1具有减轻缺氧复氧肾小管上皮细胞炎性损伤的作用,其可减少细胞分泌炎性因子,作用机制与内质网应激介导的Caspase-11途径有关。
Objective: To study the effects of SIRT1 on inflammatory injury of renal tubular epithelial cells after hypoxia reoxygenation. Methods: Construction of hypoxia reoxygenation injury model of renal tubular epithelial cells.The expression of SIRT1 in cells was detected by qRT-PCR and Western blot.Transfection pcDNA3.1-SIRT1 into renal tubular epithelial cells.qRT-PCR and Western blot were used to detect the efficiency of overexpression.ELISA assay was used to detect the secretion of TNF-α and IL-1β in cells.The level of CHOP and Caspase-11 protein was detected by Western blot.The content of LDH in culture medium was detected by automatic biochemical analyzer. Results: The levels of SIRT1 mRNA and protein were decreased in anoxia reoxygenation renal tubular epithelial cells.After transfection,the levels of SIRT1 mRNA and protein in pcDNA3.1-SIRT1 cells increased significantly.The levels of TNF-α and IL-1β secreted by anoxia reoxygenation renal tubular epithelial cells increased,the levels of CHOP and Caspase-11 protein in the cells increased.The level of LDH increased in the cell culture medium.After overexpression of SIRT1,renal tubular epithelial cells were treated with anoxia and reoxygenation.The levels of TNF-α and IL-1β secreted by the cells decreased,the level of CHOP and Caspase-11 protein in the cells decreased,the level of LDH in cell culture medium decreased. Conclusion: SIRT1 can alleviate inflammatory injury of renal tubular epithelial cells after hypoxia reoxygenation,it can reduce the secretion of inflammatory factors in cells,the mechanism is related to endoplasmic reticulum stress mediated Caspase-11 pathway.
作者
俞济荣
刘璘琛
许圣淳
王凤梅
YU Ji-Rong;LIU Lin-Chen;XU Sheng-Chun;WANG Feng-Mei(Department of Nephrology,Zhongda Hospital Southeast University,Nanjing 210009,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第9期1036-1041,共6页
Chinese Journal of Immunology
关键词
肾小管上皮细胞
炎症因子
SIRT1
缺氧复氧
Renal tubular epithelial cells
Inflammatory factors
SIRT1
Hypoxia reoxygenation
