基于NLRP3/Caspase-1/IL-1β炎症通路探讨越婢汤和真武汤对阿霉素肾病大鼠的影响

Exploration on Effects of Yuebi Decoction and Zhenwu Decoction on Adriamycin Nephropathy in Rats Based on NLRP3/caspase-1/IL-1βInflammatory Pathway

目的:同时观察2个不同利水经方越婢汤及真武汤对阿霉素肾病模型大鼠肾脏组织中Nod-like Receptor Pyrin Domain Containing 3(NLRP3)、半胱天冬酶(cysteinyl aspartate specific proteinase,Caspase)-1及其下游炎症因子白细胞介素(interleukin,IL)-1β蛋白表达的影响。方法:选用雄性SD大鼠随机分为正常组、模型组、越婢汤组、真武汤组每组各12只,适应性喂养1周后,2次(间隔1周)尾静脉注射阿霉素,于第2次注射2周后开始越婢汤及真武汤灌胃给药,6周后留取24 h尿蛋白定量(24-hour urine protein quantification,24hUTP)处死大鼠,腹主动脉取血,检测血清白蛋白(Serum albumin,ALB)、丙氨酸氨基转移酶(Alanine aminotransferase,ALT)、血肌酐(serum creatinine,Scr)、尿素氮(urea nitrogen,BUN)、胆固醇(Cholesterol,CHOL)、甘油三酯(triglyceride,TG)水平;光镜观察肾小球及肾小管间质病理改变,免疫组化观察大鼠肾脏NLRP3、Caspase-1、IL-1β的表达。结果:与正常组比较,模型组大鼠血清ALB下降,血脂水平、24hUTP、肾组织NLRP3、Caspase-1、IL-1β蛋白表达明显升高(P<0.05),大鼠肾脏出现明显病理改变,而肾功能无明显变化(P>0.05);与模型组比较,越婢汤组及真武汤组大鼠血清ALB明显升高、血脂水平及24hUTP明显下降(P<0.05),大鼠肾脏病理明显改善,肾组织NLRP3、Caspase-1、IL-1β蛋白表达明显下降(P<0.05),且越婢汤与真武汤2组之间比较各项指标差异均无统计学意义(P>0.05)。结论:越婢汤及真武汤均能通过影响阿霉素肾病模型大鼠肾组织中NLRP3、Caspase-1及IL-1β蛋白的表达,达到改善肾脏病理改变及各项生化指标的作用。

Objective:To observe the effects of two different excreting water classical prescriptions(Yuebi Decoction and Zhenwu Decoction)on the protein expression of NLRP3,Caspase-1,and IL-1βin the kidney tissue of adriamycin nephropathy rats model.Methods:SD rats were selected and randomly divided into the normal group,the model group,the Yuebi Decoction group and the Zhenwu Decoction group,12 rats were in each group.After one week's adaptive feeding,doxorubicin was injected into the tail vein twice(1 week apart).Two weeks after the second injection,rats in Yuebi Decoction group and Zhenwu Decoction group were given corresponding Chinese medicine by gavage for 6 weeks.After taking 24-hour urine protein quantification(24hUTP),the rats were sacrificed,and their bloods were taken from the abdominal aorta.The ALB,ALT,Scr,BUN,CHOL,TG levels were tested,and the light microscope was applied to observe the pathological changes of glomeruli and tubulointerstitium.Meanwhile,the expression of NLRP3,Caspase-1,IL-1βwere observed in the rats'kidney by immunohistochemistry.Results:In comparison with the normal group,the serum ALB level decreased,while the blood lipids,24hUTP,renal tissue NLRP3,Caspase-1,and IL-1βincreased significantly(P<0.05),the pathology of the rat kidney was significantly damaged in the model group.In contrast to the model group,the ALB of the Yuebi Decoction group and the Zhenwu Decoction group increased,and the 24hUTP and blood lipids decreased(P<0.05),the pathology of the rat kidney was significantly improved,and the expression of NLRP3,Caspase-1,IL-1βin the kidney tissue was significantly decreased(P<0.05).Moreover,there was no significant difference between the two groups of Yuebi Decoction and Zhenwu Decoction(P>0.05).Conclusion:The two different excreting water classical prescriptions(Yuebi Decoction and Zhenwu Decoction)can affect the expression of NLRP3,Caspase-1 and IL-1βprotein in the renal tissue of rats with adriamycin nephropathy,so as to improve the kidney pathological and biochemical indexes.