糖皮质激素受体在缺血再灌注肾损伤中的作用

doi:10.3969/j.issn.1006-7795.2022.02.017

摘要/Abstract

摘要： 目的探究糖皮质激素受体(glucocorticoid receptor, GR)在缺血再灌注(ischemia reperfusion, IR)诱导的小鼠肾损伤过程中的变化及作用。方法使用微血管夹夹闭小鼠左侧肾蒂45 min诱导小鼠缺血再灌注肾损伤模型,分别于第5、15、30和80天收集小鼠血液,通过试剂盒检测血尿素氮(blood urea nitrogen, BUN)和血肌酐(serum creatinine, Scr)的浓度。采用过碘酸希夫染色法(periodic acid-Schiff staining, PAS)和苏木精-伊红染色(hematoxylin-eosin staining, HE)观察肾组织炎性细胞浸润情况;通过免疫荧光(immunofluorescence, IF)法检测肾损伤分子-1(kidney injury molecular-1, Kim-1)、炎症细胞标志物F4/80和CD3的表达变化;通过实时荧光定量PCR(real time quantitative PCR,RT-qPCR)、IF和蛋白质印迹技术(Western blotting, WB)检测GR的表达。选取8周龄雌性小鼠肾脏单侧IR后进行腹腔注射米非司酮(mifepristone, MF, 50 mg·kg^-1·d^-1),检测实验组和对照组的BUN和Scr浓度,IF检测Kim-1和F4/80的蛋白水平。结果与未处理组相比,小鼠缺血再灌注肾损伤后,第5天小鼠血清BUN升高4.9倍,Scr升高8.3倍;RT-qPCR结果显示,IR组小鼠炎症分子的mRNA较未处理空白组升高3~5倍,GR的mRNA较空白组下降3.5倍。IF和WB结果显示,GR的蛋白质水平显著下调;IR处理的同时,注射MF阻断GR后,BUN和Scr较注射玉米油的对照组均增高,表明阻断GR会导致IR后肾功能损伤加重,IF结果显示Kim-1和F4/80表达明显升高,说明炎性反应增强,肾损伤加重。结论 GR的表达和IR诱导肾损伤存在显著的相关性,同时阻断GR会进一步加重IR诱导的肾损伤。

关键词: 肾损伤, 糖皮质激素受体, 缺血再灌注, 炎症反应, 米非司酮

Abstract: Objective To investigate the changes and role of glucocorticoid receptor(GR) in ischemia-reperfusion(IR)-induced kidney injury. Methods The left renal pedicle of the mouse was clamped for 45 minutes with a microvascular clip to induce kidney injury.The blood urea nitrogen (BUN) and serum creatinine (Scr) levels were detected with the BUN and Scr kits on the 5th,15th,30th and 80th day. Periodic acid-Schiff (PAS) staining and hematoxylin-eosin (HE) staining were used to measure the inflammatory cells infiltration of renal tissues.The expression levels of kidney injury molecular-1 (Kim-1), inflammatory cell markers F4/80 and CD3 were disclosed by immunofluorescence (IF). The mRNA level of GR was detected with real time quantitative PCR (RT-qPCR), and its protein level was measured with IF and Western blotting(WB) technology. After intraperitoneal injection of mifepristone (MF, 50 mg·kg^-1·d^-1) in IR-induced 8 weeks female mice, the levels of BUN and Scr in the experimental group and the control group were measured, and the expression of Kim-1 and F4/80 were detected with IF. Results BUN and Scr level of the IR group was 4.9 and 8.3 times higher than that of the sham group in 5th day after IR.The mRNA level of inflammatory cells markers in the IR group was 3-5 times higher, respectively, than those of the sham group, and the mRNA level of GR was 3.5 times lower than those of the sham group. The protein level of GR also dropped significantly. At the same time of IR treatment, after injection of MF to block GR, BUN and Scr were increased compared with the control group, indicating that blocking GR would lead to aggravated renal function damage after IR. The expression levels of Kim-1 and F4/80 in the experimental group were prominently upregulated, which indicated that when GR was blocked,the inflammatory response was enhanced and the kidney injury was aggravated. Conclusion This study revealed that there was a significant correlation between the expression of GR and the degree of renal injury in IR induced renal injury and blocking GR could aggravate IR-induced renal injury.

Key words: kidney injury, glucocorticoid receptor(GR), ischemia-reperfusion(IR), inflammatory response, mifepristone

中图分类号:

王向东, 姜敏, 王鑫, 巩永凤. 糖皮质激素受体在缺血再灌注肾损伤中的作用[J]. 首都医科大学学报, 2022, 43(2): 260-268.

Wang Xiangdong, Jiang Min, Wang Xin, Gong Yongfeng. The role of glucocorticoid receptor in ischemia-reperfusion-induced kidney injury[J]. Journal of Capital Medical University, 2022, 43(2): 260-268.

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