摘要
目的:探讨维生素D(VD)在肾脏缺血再灌注(I/R)损伤中的作用及相关机制。方法:将48只雄性C57BL/6J小鼠分为4组:假手术组(Sham组)、活性VD类似物阿法骨化醇处理组(VD组)、肾缺血再灌注损伤组(I/R组)、阿法骨化醇处理的I/R组(I/R+VD组)。I/R损伤造模24 h后处死各组小鼠,收集外周血,检测血尿素氮(BUN)、血清肌酐(SCr)和炎症因子IL-1β、IL-18、肿瘤坏死因子-α(TNF-α)含量;收集各组小鼠肾组织,TUNEL染色法检测各组小鼠肾组织细胞凋亡水平,免疫组织化学检测肾组织中NOD样受体蛋白3(NLRP3)的表达阳性率;Western blot检测各组小鼠肾组织中NLRP3炎症小体相关因子NLRP3、GSDMD-N、cleaved Caspase-1及NF-κB p65、IκBα的蛋白表达水平。结果:与Sham组比较,VD组血清中BUN、SCr和IL-1β、IL-18及TNF-α水平差异无统计学意义(P>0.05),I/R组与I/R+VD组明显升高(P<0.05);与I/R组比较,I/R+VD组BUN、SCr和IL-1β、IL-18及TNF-α水平显著降低(P<0.05);与Sham组比较,VD组小鼠肾组织中TUNEL阳性率与NLRP3表达差异无统计学意义(P>0.05),I/R组与I/R+VD组明显升高(P<0.01),与I/R组比较,I/R+VD组的TUNEL阳性率与NLRP3表达均显著降低(P<0.05);Western blot检测结果显示,与Sham组相比,VD组小鼠肾组织中NLRP3、GSDMD-N、cleaved Caspase-1、IL-1β和NF-κB p65、IκBα的蛋白表达水平差异无统计学意义(P>0.05),I/R组与I/R+VD组除IκBα蛋白表达明显降低外(P<0.05),其他蛋白表达水平均明显升高(P<0.05);与I/R组比较,I/R+VD组中NLRP3、GSDMD-N、cleaved Caspase-1和IL-1β和NF-κB p65蛋白表达水平均明显降低(P<0.05),IκBα蛋白表达水平明显降低(P<0.05)。结论:VD可通过抑制NF-κB途径介导的NLRP3炎症小体激活,减轻I/R损伤的炎症反应,发挥肾脏保护作用。
Objective:To investigate the role and mechanism of vitamin D(VD)in renal ischemia-reperfusion(I/R group)injury.Methods:Forty-eight male C57BL/6J mice were divided into four groups:Sham operation group(Sham group),active VD analog alfacalcidol treatment group(VD group),renal ischemia-reperfusion injury group(I/R group)and alfacalcidol treated I/R group(I/R+VD group).After 24 h renal I/R injury,mice in each group were sacrificed,and peripheral blood was collected to detect the levels of blood urea nitrogen(BUN),serum creatinine(SCr)and inflammatory factors IL-1β,IL-18 and tumor necrosis factor-α(TNF-α).The renal tissues of mice in each group were collected,the apoptosis level of renal tissues was detected by TUNEL staining,and the positive expression rate of NLRP3 was detected by IHC.Western blot was used to detect the NLRP3 inflammasome associated factors NLRP3,GSDMD-N,cleaved Caspase-1,IL-1βand NF-κB p65,IκBαin renal tissues of mice in each group.Results:Compared with Sham group,there was no significant difference in serum BUN,SCr,IL-1β,IL-18 and TNF-αlevels in VD group(P>0.05),significantly increased in I/R group and I/R+VD group(P<0.05).Compared with I/R group,BUN,SCr,IL-1β,IL-18 and TNF-αwere significantly decreased in I/R+VD group(P<0.05).Compared with Sham group,the positive rate of TUNEL and NLRP3 expression in renal tissues of mice in VD group had no significant difference(P>0.05),the positive rate of TUNEL and NLRP3 expression in renal tissues of mice in I/R group and I/R+VD group were significantly increased(P<0.01),while the positive rate of TUNEL and NLRP3 expression in I/R+VD group were significantly decreased(P<0.05).Western blot results showed that compared with Sham group,there was no significant difference in protein expression levels of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β,NF-κB P65,IκBαin VD group(P>0.05).The protein expression of IκBαin I/R group and I/R+VD group was significantly decreased(P<0.05),while the other protein expression levels were significantly increased(P<0.05).Compared with the I/R group,the expression levels of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1βand NF-κB p65 were significantly decreased in the I/R+VD group(P<0.05),while the expression level of IκBαwas significantly decreased(P<0.05).Conclusion:VD plays a protective role in I/R injury by inhibiting NF-κB mediated activation of NLRP3 inflammasome.
作者
王平
李辉
谢枫
苑晓姣
WANG Ping;LI Hui;XIE Feng;YUAN Xiaojiao(Department of Critical Care Medicine,Xinjiang Production and Construction Corps Hospital,Urumqi 830002,China;Department of Oncology,Xinjiang Production and Construction Corps Hospital,Urumqi 830002,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第7期1381-1386,共6页
Chinese Journal of Immunology
基金
兵团科技攻关项目(2018AB024)。
