缺氧复氧诱导大鼠肾小管上皮细胞内质网应激反应 - 第三军医大学学报.doc

内质网应激反应(400042 重庆，第三军医大学大坪医院野战外科研究所肾内科) [摘要] 目的: 观察大鼠肾小管上皮细胞NRK-52E细胞）缺氧复氧时内质网应激蛋白表达的改变及其意义,以探讨缺氧复氧内质网应激反应方法: NRK-52E5%胎牛血清的DMEM液培养于37℃、5%CO2 恒温培养箱。将细胞分为正常对照组和缺氧/复氧损伤（hypoxia reoxygenation，H/R）组。采用全自动生化分析仪检测细胞培养液上清LDH活性，Western blot方法GRP78、CHOP蛋白表达结果: 22.07±1.23）U/L]比较，H/R组[（45.93±6.42）U/L]显著增加(P 0.05)。②与对照组肾小管上皮细胞GRP78表达(0.80±0.04)比较, H/R组(1.24±0.04)明显增加(P 0.05)；与对照组肾小管上皮细胞CHOP表达(0.81±0.05)比较, H/R组 (1.19±0.05)明显增加(P 0.05)。③与对照组IL-1β水平（6.55±2.03）比较，H/R组（14.93±2.26）显著增加(P 0.01)。结论: 缺氧复氧可以诱导大鼠肾小管上皮细胞内质网应激。[关键词] 内质网应激Role of endoplasmic reticulum stress in hypoxia/reoxygenation induced inflammation in rat renal tubular epithelial cells Tong Yuna, Yang Jurong , Jing Yu，Zhang Jianguo，Li Kailong，HE Yani (Department of Nephrology, Institute of Surgery Research, Daping Hospital,Third Military Medical University, Chongqing 400042, China) [Abstract] Objective To investigate the effect of hypoxia / reoxygenation on endoplasmic reticulum stress（ERS）in rat renal tubular epithelial cells（NRK-52E cells）.Methods NRK-52E cells in 10% FBS DMEM culture medium were divided into 2 groups , control group and hypoxia / reoxygenation (H/R) group. Cells in H/R group were exposed to hypoxia for 4 hours and subsequently reoxygenation for 12 hours. LDH in culture supernatant was detected by automatic biochemistry analyzer. Western blot was applied to detect the expression of GRP78 and CHOP. IL-1β in culture medium was detected by ELISA. Results Cell viability was decreased obviously after hypoxia /reoxygenation (P 0.05). Compared with untreated cells, the expression of GRP78 and CHOP protein of the cells had increased significantly in the hypoxia / reoxygenation cells (P 0.05). The expression of IL-1βincreased distinctly in the hypoxia / reoxygenation cells(P 0.01).Conclusion Hypoxia / reperfusion can induce endoplasmic reticulum stress and inflammatory response in rat renal tubular epithelial cells. Endoplasmic reticulum stress may be essential in the inflammation of renal tubular e