目的 通过观察丹参多酚酸盐对核苷酸结合寡聚结构域样受体蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)/天冬氨酸特异性半胱氨酸蛋白酶 1(Cysteinyl aspartate specific proteinase-1,Caspase-1)/焦亡效应蛋白消皮素 D(Gasdermin-D,GSDMD)细胞焦亡通路的影响,探讨改善膜性肾病大鼠肾损伤可能的作用机制.方法 SD大鼠通过尾静脉注射阳离子牛血清白蛋白建立膜性肾病大鼠模型,造模成功后分为模型组、盐酸贝那普利组(10 mg/kg)、丹参多酚酸盐低、中、高剂量组(16.7、33.3、66.7 mg/kg),另设有正常组,每组均为10只.各给药组连续给药4周后,检测24小时尿蛋白定量(24-hour urinary protein quantity,24 h UTP)和血肌酐(serum creatinine,Scr)、血尿素氮(blood urea nitrogen,BUN)、血清总蛋白(total protein,TP)、血清白蛋白(serum albumin,ALB)、甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)的含量;酶联免疫吸附法检测大鼠血清白细胞介素-1β(interleukin-1β,IL-1β)、IL-18;采用光镜、电镜、荧光显微镜下观察肾脏病理学变化;Western blot、实时荧光定量PCR法检测NLRP3、Caspase-1、GSDMD蛋白及mRNA表达.结果 与模型组比较,各给药组24 h UTP水平明显降低(P＜0.05,P＜0.01),丹参多酚酸盐中、高剂量组和盐酸贝那普利组大鼠血清TP和ALB水平显著升高(P＜0.01),TC和TG水平明显降低(P＜0.05,P＜0.01);普通光镜、荧光显微镜及电镜下观察可见模型组大鼠肾组织病理损伤明显,经各组药物治疗后病理损伤逐渐改善.与模型组比较,各给药组大鼠血清IL-1β、IL-18水平明显降低(P＜0.05,P＜0.01);与正常组比较,模型组大鼠肾脏NLRP3、Caspase-1、GSDMD蛋白及其mRNA表达显著升高(P＜0.01);与模型组比较,各给药组大鼠肾组织NLRP3/Caspase-1/GSDMD蛋白及其mRNA表达水平明显降低(P＜0.05,P＜0.01),以丹参多酚酸盐高剂量组效果最佳.结论 丹参多酚酸盐对膜性肾病大鼠肾保护作用与调控NLRP3/Caspase-1/GSDMD细胞焦亡通路,改善炎症状态,减轻肾损伤有关.
Effects of salvianolate on renal injury in rats with membranous nephropathy through NLRP3/Caspase-1/GSDMD pyroptosis pathway
Objective To explore the possible mechanism of Salvianolate in improving renal injury in rats with membranous nephropathy(MN),the effects of salvianolate on NOD-like receptor thermal protein domain associated protein 3(NLRP3)/Cysteinyl aspartate specific proteinase-1(Caspase-1)/Gasdermin-D(GSDMD)were observed.Methods SD rats were injected with cationic bovine serum albumin(C-BSA)through the tail vein to establish a MN rat model.After the model was established suc-cessfully,they were divided into the model group,the benazepril hydrochloride group(10 mg/kg),the low,medium,and high dose groups of salvianolic acid(16.7,33.3,66.7 mg/kg),and the normal group with 10 rats in each group.After successful modeling,the levels of 24-hour urinary protein quantification(24 h UTP),blood creatinine(SCR),blood urea nitrogen(BUN),total protein(TP),albumin(ALB),triglycerides(TG),and total cholesterol(TC)were measured in each treatment group after continuous administration for 4 weeks;the method of ELISA was used to detect interleukin-1 in rats β(IL-1)β、interleukin-18(IL-18);the pathological changes of kidney were observed under light microscope,electron microscope and fluorescence microscope.Western blot and real-time fluorescence quantitative PCR were used to detect the expression of NLRP3,Caspase-1,GSDMD proteins and mRNA.Results Compared with the model group,the 24 h UTP levels of each treatment group were significantly decreased(P＜0.05,P＜0.01),while the levels of TP and ALB of rats in the middle and the high dose groups of salvianolic acid salts and the benazepril group were significantly increased(P＜0.01),and the levels of TC and TG were significantly decreased(P＜0.05,P＜0.01).Under ordinary light microscope,fluorescence microscope and electron microscope,it was observed that the pathological damage of renal tissue in the model group was obvious,and the pathological injury was gradually improved after drug treatment in each group.Compared with the model group,the level of IL-1 β of rats in each treatment group.The level of IL-18 was significantly decreased(P＜0.05,P＜0.01).The expression of NLRP3,Caspase-1,GSDMD proteins and their mRNA in the kidneys of the model group rats was significantly increased(P＜0.01);the expression levels of NLRP3/Caspase-1/GSDMD protein and its mRNA in the renal tissue of rats in each treatment group were significantly decreased(P＜0.05,P＜0.01),with the highest dose of salvianolic acid salt group showing the best effect.Conclusion The protective effect of salvianolate on membranous nephropathy may be related to the regulation of NLRP3/Caspase-1/GSDMD cell pyrogenesis pathway and the reduction of inflammatory renal injury.
万方数据
维普
