受体交互作用蛋白激酶1抑制剂necrostatin-1治疗狼疮性肾炎的实验研究

Experimental study of receptor interaction protein kinase 1 inhibitor(necrostatin-1)in treatment of lupus nephritis

目的:探讨狼疮性肾炎(LN)中坏死性凋亡的机制和受体交互作用蛋白激酶1(receptor interaction protein kinase 1,RIPK1)抑制剂necrostatin-1(Nec-1)对疾病的疗效。方法:体内实验应用MRL/lpr自发诱导的系统性红斑狼疮小鼠,19周龄时腹腔注射RIPK1抑制剂Nec-1[3.5mg/(kg·d)]3周。检测血清肌酐、补体水平和尿蛋白肌酐比值变化,观察肾脏病理改变,检测坏死性凋亡相关蛋白表达变化。体外实验检测Nec-1处理系膜细胞坏死性凋亡的变化,利用qRT-PCR检测促炎因子表达变化。结果:经Nec-1治疗后,LN小鼠全身免疫炎症反应和肾脏损伤缓解,血清抗核抗体、抗双链DNA抗体滴度和尿蛋白,肌酐比值降低。体外实验发现,Nec-1抑制系膜细胞坏死性凋亡和促炎因子释放。结论:RIPK1抑制剂Nec-1可抑制系膜细胞的坏死性凋亡,改善LN的症状。

Objective:To explore the mechanism of necroptosis in lupus nephritis and the efficacy of receptor interaction protein kinase 1(RIPK1)inhibitor necrostatin-1(Nec-1)in the treatment of lupus nephritis.Methodology:MRL/lpr mice at the age of 19 weeks was injected intraperitoneally with Nec-1[3.5 mg/(kg·d)]for 3 weeks.Changes were detected in serum creatinine,complement levels and urine protein/creatinine ratio.Kidney pathological staining was used to observe kidney pathological changes and the changes in expression of necroptosis-related proteins.In vitro,the expression of necroptosis-related molecules in mesangial cells treated with Nec-1 were measured.Changes in the expression of pro-inflammatory factors were detected by qRT-PCR.Results:The systemic immune inflammatory response and kidney damage of mice with lupus nephritis were alleviated after treatment with Nec-1,serum anti-nuclear antibodies titers,anti-double-stranded DNA antibody titers,and urine protein/creatinine ratio were decreased,and Nec-1 inhibited mesangial cell necrosis and the release of pro-inflammatory factors in vitro.Conclusion:RIPK1 inhibitor Nec-1 can inhibit the necrotizing apoptosis of mesangial cells and relieve the symptoms of lupus nephritis.