基于TXNIP/NLRP3/Caspase-1通路探讨清热止血方联合雷公藤多苷治疗过敏性紫癜性肾炎模型大鼠的作用机制  

Based on TXNIP/NLRP3/Caspase-1 pathway to explore the mechanism of Qingre Zhixue Formula combined with tripterygium wilfordii polyglycosides in the treatment of henochschönlein purpura nephritis in rats

作　　者：王龙[1,2] 丁樱 徐闪闪[2] 代彦林 张霞 WANG Long;DING Ying;XU Shanshan;DAI Yanlin;ZHANG Xia(School of Pediatrics,Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450003,China)

机构地区：[1]河南中医药大学儿科医学院,郑州450046 [2]河南中医药大学第一附属医院

出　　处：《北京中医药大学学报》2023年第4期476-483,共8页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(No.81873343);河南省中医药科学研究专项(No.2023ZY2044)。

摘　　要：目的 探讨清热止血方联合雷公藤多苷对过敏性紫癜性肾炎模型大鼠的作用机制。方法 腹腔注射卵白蛋白与完全弗氏佐剂复制过敏性紫癜性肾炎大鼠模型。将造模成功的54只大鼠按照随机数字表法分为模型组(n=11)、清热止血方组(3.17 mg/kg,n=11)、雷公藤多苷组(4.69 mg/kg,n=11)、联合组(清热止血方3.17 mg/kg+雷公藤多苷4.69 mg/kg,n=11)、激素组(醋酸泼尼松2.34 mg/kg,n=10),另设空白组(n=9)。各给药组给予相应药物灌胃,空白组及模型组予等量的生理盐水灌胃,2次/d,连续灌胃4周。采用尿液分析仪检测大鼠尿红细胞计数及24 h尿蛋白定量;采用透射电镜观察大鼠肾小球系膜区病变情况;采用蛋白质印迹法及实时荧光PCR法检测大鼠肾脏组织硫氧还蛋白相互作用蛋白(TXNIP)、NOD样受体蛋白3(NLRP3)、半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)、白细胞介素-1β(IL-1β)的蛋白及mRNA表达量。结果 与模型组比较,各给药组大鼠尿红细胞计数,24 h尿蛋白定量,肾脏组织中TXNIP、NLRP3、Caspase-1蛋白表达量均降低(P<0.05);各给药组大鼠肾小球系膜增生及基底膜增厚均改善,其中,联合组改善最明显;联合组、雷公藤多苷组及激素组大鼠肾脏组织中TXNIP、NLRP3 mRNA表达量降低(P<0.05),联合组、激素组大鼠肾脏组织中IL-1β mRNA表达量降低(P<0.05)。与联合组比较,其余给药组大鼠尿红细胞计数,24 h尿蛋白定量,肾脏组织中TXNIP、NLRP3蛋白及IL-1β mRNA表达量均升高(P<0.05);雷公藤多苷组大鼠肾脏组织中TXNIP、NLRP3 mRNA表达量升高(P<0.05),清热止血方组大鼠肾脏组织中NLRP3 mRNA表达量升高(P<0.05),激素组大鼠肾脏组织中TXNIP、NLRP3、IL-1β mRNA表达量均升高(P<0.05)。结论 清热止血方联合雷公藤多苷可有效降低过敏性紫癜性肾炎模型大鼠尿红细胞计数及24 h尿蛋白定量,减轻肾脏损伤,可能与抑制TXNIP/NLRP3/Caspase-1炎性通路的激活相关。Objective We aimed to explore the therapeutic mechanism of Qingre Zhixue Formula combined with tripterygium wilfordii polyglycosides on rats with henoch-schönlein purpura nephritis(HSPN).Methods The model rats of HSPN were established.According to the random number table method,54 rats were divided into model group(n=11),Qingre Zhixue Formula group(3.17 mg/kg,n=11),tripterygium wilfordii polyglycoside group(4.69 mg/kg,n=11),combined group(Qingre Zhixue Formula 3.17 mg/kg+tripterygium wilfordii polyglycoside tablets 4.69 mg/kg,n=11),hormone group(prednesone acetate tablets 2.34 mg/kg,n=10),and the blank group(n=9)was set.The blank group and model group were gavaged the same amount of physiological saline,and the other treatment groups were gavaged with corresponding drugs twice a day for 4 weeks.Urine red blood cell count and 24 hours urine protein quantification were detected by urine analyzer.The pathological changes of mesangial area of rats were observed by transmission electron microscope.The expressions of thioredoxin interacting protein(TXNIP),Nod-like receptor protein 3(NLRP3),cysteine aspartic protease-1(Caspase-1)and interleukin-1β(IL-1β)protein and mRNA in rat kidney tissue were detected by Western blotting and real-time PCR.Results Compared with the model group,the urine red blood cell count,24 hours urine protein quantification,and the expression of TXNIP,NLRP3 and IL-1βprotein in the kidney tissues of rats in each treatment group were all reduced(P<0.05);the glomerular thylakoid hyperplasia and basement membrane thickening of rats in each treatment group were improved,among which the most obvious improvement was observed in the combined group;The mRNA expression of TXNIP and NLRP3 in the kidney tissues of the rats in the combined group,the tripterygium wilfordii polyglycoside group and the hormone group were decreased(P<0.05),and the mRNA expression of IL-1βin the kidney tissues of the rats in the combined group and the hormone group decreased(P<0.05).Compared with the combined group, the urine

关 键 词：过敏性紫癜性肾炎 雷公藤多苷 清热止血方 硫氧还蛋白相互作用蛋白/NOD样受体蛋白3/半胱氨酸天冬氨酸蛋白酶-1通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]