【摘要】 目的：探讨核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体在特发性膜性肾病（IMN）中的作用及其激活的可能机制。方法：收集未行肾上腺皮质激素及免疫抑制剂治疗且肾组织活检诊断为IMN的患者135例，10例因肾结核及肾肿瘤行肾切除的患者肾脏组织作为对照组。免疫组化法检测各肾组织样本NLRP3、肿瘤坏死因子受体相关因子6(TRAF6)、Caspase-1、NF-κB p65、MAPK信号通路相关蛋白磷酸化ERK1/2(p-ERK1/2)、磷酸化p38(p-p38)及磷酸化JNK(p-JNK)因子表达；收集IMN患者血液和24 h尿液，分析24 h尿蛋白定量、血肌酐、肌酐清除率、尿素氮等一般临床资料，对尿蛋白与NLRP3、Caspase-1、IL-1β、TNF-α及NLRP3与TRAF6进行相关性分析。结果：与对照组比较，IMN患者24 h尿蛋白和血肌酐明显升高（均P<0.05），肾组织NLRP3和Caspase-1蛋白表达明显升高（均P<0.05），其中Ⅱ期IMN患者肾组织NLRP3和Caspase-1蛋白表达最高，IL-1β和TNF-α表达增强（均P<0.05），其中Ⅱ期IMN患者肾组织IL-1β和TNF-α蛋白表达最强，NF-κB p65、p-ERK1/2、p-p38和p-JNK蛋白表达升高（均P<0.05）。IMN患者24 h尿蛋白与肾组织NLRP3蛋白表达呈正相关（r=0.689,P<0.01），与Caspase-1蛋白呈正相关（r=0.614,P<0.000 1），与IL-1β呈正相关（r=0.708,P<0.000 1），与TNF-α呈正相关（r=0.594,P<0.01）。结论：TRAF6通过NF-κB激活诱导ERK1/2、p38、JNK信号通路刺激NLRP3炎症小体活化，促进IMN发生和大量尿蛋白产生。更多还原

【Abstract】 Objective:To explore role of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) inflammasome in idiopathic membranous nephropathy(IMN) and its possible mechanism of activation. Methods:A total of 135 patients with no adrenal corticosteroid and immunosuppressive therapy who were diagnosed with IMN by renal biopsy were collected, and 10patients with renal tuberculosis and renal tumors who underwent nephrectomy were selected as control group. Immunohistochemical was used to detect NLRP3, tumor necrosis factor receptor-related factor 6(TRAF6), Caspase-1, NF-κB p65, MAPK signaling pathway-related proteins, including phosphorylated ERK1/2(p-ERK1/2), phosphorylated p38(p-p38), phosphorylated JNK(p-JNK) expressions. Blood and 24-hour urine of IMN patients were collected, and 24-hour urine protein quantification, serum creatinine, creatinine clearance rate, blood urea nitrogen and other general clinical data were were analyzed, correlation between urine protein and NLRP3, Caspase-1, IL-1β, TNF-α, NLRP3 and TRAF6 were analyzed, respectively. Results:Compared with control group, 24-hour urine protein and serum creatinine of IMN patients were significantly increased(both P<0.05), and protein expressions of NLRP3 and Caspase-1 in kidney tissues of IMN patients were significantly increased(both P<0.05), among which renal tissue of stage Ⅱ IMN patients had the highest protein expressions of NLRP3 and Caspase-1, IL-1β and TNF-α expressions were enhanced(both P<0.05), and IL-1β and TNF-α protein expressions in renal tissue of stage Ⅱ IMN patients were the strongest, protein expressions of NF-κB p65, p-ERK1/2, p-p38 and p-JNK were significantly increased(all P<0.05). 24-hour urinary protein of IMN patients was positively correlated with expression of NLRP3 protein in kidney tissues(r=0.689, P<0.01), positively correlated with Caspase-1protein(r=0.614, P<0.000 1), positively correlated with IL-1β(r=0.708, P<0.000 1), positively correlated with TNF-α(r=0.594, P<0.01). Conclusion:TRAF6 induces ERK1/2, p38 and JNK signaling pathways to stimulate activation of NLRP3 inflammasome through activation of NF-κB, which promotes occurrence of IMN and production of large amounts of urine protein.更多还原