脂联素对脓毒症小鼠急性肾损伤的治疗作用及机制

Therapeutic effect of adiponectin on acute renal injury in mice with sepsis and its mechanism

目的 探究脂联素(APN)对脓毒症小鼠急性肾损伤的治疗作用及相关机制。方法 选取雄性C57BL/6小鼠24只,采用随机数字表法将小鼠分为空白对照组(CON组)、模型组(LPS组)、实验组(LPS+APN组)。按10 mg/kg腹腔注射脂多糖(LPS)制备脓毒症模型,CON组腹腔注射等量0.9%氯化钠注射液,实验组在造模前8 h按0.1 mg/kg尾静脉注射APN进行干预。造模后24 h处死小鼠,取静脉血检测肾损伤指标肌酐、尿素氮;HE染色观察和评估小鼠肾组织病理变化;超氧化物阴离子荧光探针(DHE)检测活性氧含量;实时定量PCR法检测肾组织中NLRP3、IL-1β和TNF-α的mRNA表达,Western blotting法检测NLRP3、Caspase-1和IL-1β蛋白表达。结果 与模型组比较,经APN预处理后,小鼠肾功能及组织病理学损伤减轻,肾组织活性氧含量明显减少,NLRP3、IL-1β和TNF-α的mRNA表达降低,NLRP3、Caspase-1和IL-1β蛋白表达下降(P均<0.05)。结论 APN对小鼠脓毒症急性肾损伤具有治疗作用,其机制可能与NLRP3炎症小体及细胞焦亡相关通路受抑制有关。

Objective To explore the effect of adiponectin(APN) on acute kidney injury in mice with sepsis and its related mechanism.Methods Twenty-four male C57BL/6 mice were randomly divided into the control group(CON group),model group(LPS group) and experimental group(LPS+APN group).We prepared the sepsis models by intraperitoneal injection of 10 mg/kg lipopolysaccharide(LPS).Mice in the CON group were injected with equal volume of 0.9%sodium chloride injection intraperitoneally,and mice in the experimental group were injected with 0.1 mg/kg APN via the tail vein at 8 h before modeling for intervention.Mice were sacrificed at 24 h after modeling,and venous blood was taken to detect renal injury indicators such as creatinine and urea nitrogen.HE staining was used to observe and evaluate the pathological changes of mouse renal tissues.The content of reactive oxygen species was detected by dihydroethidium(DHE).The mRNA expression levels of NLRP3,IL-1β and TNF-α in the renal tissues were detected by real-time quantitative PCR,and the expression levels of NLRP3,Caspase-1 and IL-1β were detected by Western blotting.Results Compared with the model group,after APN pretreatment,renal function and histopathological injury were alleviated,the content of reactive oxygen species in renal tissues significantly decreased,the mRNA expression levels of NLRP3,IL-1βand TNF-α significantly decreased,and the expression levels of NLRP3,Caspase-1 and IL-1β significantly decreased(all P<0.05).Conclusion APN has a protective effect on acute renal injury mice with sepsis,and its mechanism may be related to the inhibition of NLRP3 inflammatome and pyroptosis-related pathway.