摘要
目的探索硫氧还蛋白相互作用蛋白(thioredoxin interaction protein,TXNIP)/NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)通路在小鼠肾缺血-再灌注损伤(ischemiareperfusion injury,IRI)后肾间质纤维化中的作用。方法选择6~8周龄、SPF级雄性C57BL/6小鼠和相同遗传背景的TXNIP基因敲除小鼠,野生型小鼠分为假手术组(Sham)、肾IRI组(IRI),TXNIP基因敲除小鼠分为Sham+TXNIP KO组和IRI+TXNIP KO组,每组12只小鼠。夹闭双侧肾蒂45 min后恢复灌注制备肾IRI模型,假手术模型仅游离双侧肾动脉,不进行其他处理。在恢复灌注后第1天、第7天和第28天检测血肌酐、尿素氮、尿肾损伤分子-1(kidney injury molecule-1,Kim-1)和中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase-associated lipocalin,NGAL)、血转化生长因子-β(transforming growth factor-β,TGF-β)、白介素(Interleukin,IL)-6;第1天和第28天取肾皮质,进行Masson染色,肾小管-间质损伤评分;qPCR检测肾皮质TGF-β、IL-6 mRNA表达;Western Blot检测TXNIP、NLRP3、Pro-IL-1β、IL-1β、α-SMA蛋白表达;ELISA检测组织MDA和SOD水平。正态分布计量资料统计前行方差齐性检验,多组间比较采用单因素方差分析,两组间比较采用LSD-t检验。结果在IRI后第1、7和28天,与Sham组比较,IRI组血肌酐、尿素氮、尿Kim-1和NGAL、血TGF-β、IL-6均持续升高(均P<0.05)。在IRI后第28天,IRI组肾间质可见大片蓝染的胶原纤维、炎症细胞浸润,肾小管损伤和肾间质纤维化评分明显高于第1天(P<0.05)。在IRI后第1、7和28天,与IRI组比较,IRI+TXNIP KO组血肌酐、尿素氮、尿Kim-1和NGAL、血TGF-β、IL-6均明显降低(均P<0.05)。在IRI后第1天和第28天,与IRI组相比,IRI+TXNIP KO组肾间质蓝色纤维组织和炎症细胞浸润减轻,肾小管损伤评分(分)[第1天(192.2±62.4)vs.(103.2±49.1),第28天(154.3±93.6)vs.(64.3±24.8),均P<0.05]和间质纤维化评分(分)[第1天(7.3±3.2)vs.(4.8±1.7),第28天(12.8±3.9)vs.(2.
Objective To explore the role of thioredoxin interaction protein(TXNIP)/NOD-like receptor protein 3(NLRP3)pathway in renal interstitial fibrosis induced by renal ischemia-reperfusion injury(IRI)in mice.Methods Adult male C57BL/6J mice aged 6 to 8 weeks and TXNIP knockout mice with the same genetic background were selected.The wild type mice were divided into the sham operation(Sham)group and renal IRI group.The TXNIP knockout mice were divided into the sham+TXNIP KO group and IRI+TXNIP KO group,with 12 mice in each group.The model of renal ischemia-reperfusion injury was established by clamping bilateral renal pedicles for 45 min and then restoring perfusion.The sham operation model was only dissociated bilateral renal arteries without other treatment.Blood creatinine,urea nitrogen,kidney injury molecule-1(Kim-1)and neutrophil gelatinase-associated lipocalin(NGAL),blood transforming growth factor-β(TGF-β)and interleukin 6(IL-6)were measured on the 1st,7th and 28th days after reperfusion.The renal cortex was taken on the 1st and 28th days for Masson staining,in which the renal tubule-interstitial injury score was obtained.TGF-βand IL-6 mRNA expression were detected by qPCR,TXNIP,NLRP3,Pro-IL-1β,IL-1βandα-SMA protein expression were detected by Western blot,and MDA and SOD levels were detected by ELISA.Homogeneity test of variance was performed before the statistics of normal distribution measurement data,one-way ANOVA was used for the comparison between multiple groups,and LSD-t test was used for the comparison between the two groups.Results On the 1st,7th and 28th days after IRI,compared with the sham group,the Scr,BUN,Kim-1,NGAL,TGF-βand IL-6 were increased continuously in the IRI group(P<0.05).On the 28th day after IRI,large areas of collagen fibers and inflammatory cell infiltration were observed in the renal interstitium of the IRI group.In the IRI group,the scores of renal tubular injury and renal interstitial fibrosis on the 28th day were significantly higher than those on the 1st day(all P<0.05).On
作者
李冠青
张亚洲
田志
王旻
Li Guanqing;Zhang Yazhou;Tian Zhi;Wang Min(Emergency Department,Hebei Provincial People's Hospital,Shijiazhuang 050057,China;Department of Orthopedics,Hebei Provincial People's Hospital,Shijiazhuang 050057,China;Department of Neurology,Hebei Provincial People's Hospital,Shijiazhuang 050057,China)
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2023年第5期637-643,共7页
Chinese Journal of Emergency Medicine
基金
河北省医学科学研究重点课题计划(20180200)。
