Nrf2调控的炎症介质表达在肠缺血再灌注所致肾脏损伤中的作用

Role of Inflammatory Mediators Regulated by Nrf2 in Intestinal Ischemia-Reperfusion Induced Kidney Injury in Rats

目的:探讨Nrf2调控的炎症介质表达在肠缺血/再灌注所致肾脏损伤中的作用。方法:健康雄性SPF级C57BL/6J小鼠36只,随机均分为如下3组:假手术组(S组)、缺血再灌注组(I/R组)、拮抗剂Brusatol+缺血再灌注组(Brusatol+I/R组)。复制小鼠肠缺血再灌注模型,于再灌注2h时采集颈动脉血样,然后处死小鼠,取肾组织,测定血清尿素氮(BUN)、肌酐(Cr)和中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平,检测肾组织Nrf2蛋白表达,血清肿瘤坏死因子α(TNF-α)、白细胞介素-6、10(IL-6、IL-10)的含量。显微镜下观察肾组织病理学结果,并行病理学损伤评分。结果:与S组比较,I/R组肾脏组织病理学损伤评分升高(P<0.05),血清BUN、Cr和NAGL浓度升高,肾脏组织Nrf2蛋白表达上调,血清促炎因子IL-6、TNF-α在I/R组显著增高(P<0.05),抗炎因子IL-10的含量在I/R组表达显著降低(P<0.05)。使用Brusatol后,I/R组肾脏组织病理学损伤评分进一步增高(P<0.05),血清BUN、Cr和NAGL浓度进一步增高(P<0.05),肾脏组织Nrf2蛋白表达下降(P<0.05),Brusatol+I/R组血清促炎因子IL-6、TNF-α较I/R组进一步增高(P<0.05),抗炎因子IL-10的含量在I/R组表达更加降低(P<0.05)。结论:Nrf2调控的炎症介质表达在肠缺血再灌注所致肾脏损伤病程进展中的作用机制可能为调节抗炎因子/促炎因子平衡,进而调控远隔肾脏损伤。

Objective:To investigate the regulation of Nrf2 on inflammatory mediators in intestinal ischemia-reperfusion induced kidney injury in rats.Methods:Thirty-six healthy male C57BL/6J mice were randomly divided into following 3groups:sham operation group(S group),ischemiareperfusion group(I/R group),Nrf2 antagonist Brusatol+ischemia-reperfusion group(Brusatol+I/R group).The mouse model of intestinal ischemia-reperfusion was used in this study.Blood samples were collected by carotid artery 2hafter reperfusion,and the kidney tissue were collected after mice were sacrificed.Serum BUN,Cr and neutrophil gelatinase-associated lipocalin(NGAL)level were detected,and Nrf2 protein expression in renal tissue as well as serum TNF-α,IL-6and IL-10 production were also measured.Moreover,renal histopathology was then observed under microscope.Results:Histopathological kidney damage in I/R group increased significantly when compared with the S group(P<0.05).Serum BUN,Cr and NAGL concentration,Nrf2 protein expression in kidney tissue and the serum content of pro-inflammatory cytokines IL-6,TNF-αin the I/R group were significantly higher than those in the S group(P<0.05),while the serum content of anti-inflammatory cytokine IL-10 in the I/R group was obviously decreased(P<0.05).After the using of Brusatol,kidney histopathological damage score in the I/R group was further increased(P<0.05),renal tissue Nrf2 protein expression was decreased(P<0.05),while serum BUN,Cr and NAGL concentration were greatly increased than those in the I/R group without Brusatol treatment(P<0.05).In addition,serum proinflammatory cytokine IL-6,TNF-αin the Brusatol+I/R group further increased(P<0.05),and the content of anti-inflammatory cytokine IL-10 significantly decreased as compared with I/R group(P<0.05).Conclusion:Nrf2influences renal injury in intestinal ischemia reperfusion injury by regulating production of inflammatory mediators,and its mechanism may be related to the balance of anti-inflammatory/proinflammatory cytokines.