吡非尼酮通过抑制NLRP3-ASC-IL-1β通路改善大鼠肾缺血再灌注引起的急性肾损伤

Pirfenidone Improved Acute Kidney Injury by Inhibiting NLRP3-ASC-IL-1βPathway Induced by Renal Ischemia-reperfusion in Rats

目的探究吡非尼酮(PFD)对大鼠肾缺血再灌注引起的急性肾损伤的保护作用及机制。方法选取清洁级健康成年SD大鼠24只,随机分为对照组、模型组、治疗组,每组8只。模型组大鼠建立肾损伤模型,治疗组大鼠造模前给予PFD 200 mg/kg灌胃,1/d,持续1周后建立肾损伤模型。观察3组大鼠肾损伤病理变化;检测并比较3组大鼠血清中尿素、肌酐和炎症相关指标[单核细胞趋化蛋白-1(MCP-1)、白细胞介素-1β(IL-1β)]含量及MCP-1和IL-1βmRNA表达水平。检测并比较3组肾组织中核苷酸结合寡聚化结构域样受体家族结构域蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)和IL-1β蛋白相对表达量。结果与对照组比较,模型组大鼠肾脏的肾髓质高度充血,细胞核固缩、溶解、消失,肾外髓质大部分肾小管上皮细胞坏死脱落,远端肾小管及集合管内可见细胞管型。治疗组大鼠肾脏损伤较模型组减轻。与对照组比较,模型组大鼠肾损伤评分,血清中尿素、肌酐、MCP-1、IL-1β含量,MCP-1 mRNA和IL-1βmRNA表达量,NLRP3、ASC和IL-1β蛋白相对表达量均显著升高(P<0.01)。与模型组比较,治疗组大鼠肾损伤评分,血清中尿素、肌酐、MCP-1、IL-1β含量,MCP-1 mRNA和IL-1βmRNA表达量,NLRP3、ASC和IL-1β蛋白相对表达量均显著降低(P<0.05,P<0.01)。结论PFD可通过抑制NLRP3-ASC-IL-1β信号通路,降低机体炎症反应,对肾脏产生保护作用。

Objective To investigate the protective effect and mechanism of Pirfenidone(PFD)on acute kidney injury(AKI)induced by renal ischemia-reperfusion(RIR)in rats.Methods A total of 24 healthy adult SD rats of clean grade were randomly divided into control group,model group and treatment group(n=8 in each group).AKI rat models were established in model group,while rats in treatment group were given 200mg/kg Pirfenidone by intragastric administration with 1/d for a week consecutively,and then kidney injury models were established.Among 3 groups,pathological changes of kidney injury were observed,and levels of blood urea nitrogen(BUN),creatinine and contents of inflammation-related indicators[monocyte chemotactic protein 1(MCP-1)and interleukin-1β(IL-1β)]and mRNA expressions of MCP-1 and IL-1βin the serum were detected and compared.Relative expressions of nucleotide-binding oligomerization domain,leucine rich repeat and pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC)and IL-1βin renal tissues were detected and compared among 3 groups.Results Compared with those in control group,renal medulla was highly congested,and nucleus was constricted,dissolved and disappeared,while most of tubular epithelial cells in the extrarenal medulla were necrotic and detached,and cell casts were seen in distal renal tubules and collecting tubules in model group.Kidney injuries of rats in treatment group were decreased than those in model group.Compared with those in control group,score of kidney injury,contents of serum BUN,creatinine,MCP-1 and IL-1β,mRNA expressions of MCP-1 and IL-1βand relative protein expressions of NLRP3,ASC and IL-1βwere significantly increased in model group(P<0.01).Compared with those in model group,score of kidney injury,contents of serum BUN,creatinine,MCP-1 and IL-1β,mRNA expressions of MCP-1 and IL-1βand relative protein expressions of NLRP3,ASC and IL-1βwere significantly decreased in treatment group(P<0.05,P<0.01).Conclusion PFD may protect the kidney by inhibiting NLRP3-ASC-IL-1βsignaling pathway and reducing inflammatory reaction of the body.