摘要
目的:探究荆花消紫合剂对大鼠紫癜性肾炎的保护作用及对Wnt/β-catenin信号通路的作用机制.方法:选取SPF级健康雄性SD大鼠60只,随机抽取10只作为正常组,余下50只运用牛血清白蛋白(BSA)、脂多糖(LPS)和四氯化碳(CCl4)联合给药方法建立实验性大鼠IgA肾病模型,将造模成功的IgA肾病模型大鼠随机分为模型组,中药高、中、低剂量组[荆花消紫合剂给药剂量分别为10.5 mL/(kg·d)、7.0 mL/(kg·d)、3.5 mL/(kg·d)],以及西药组[阿魏酸哌嗪片13.5mg/(kg·d)],每组10只,连续给药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组织病变情况;采用蛋白质印迹法检测大鼠肾脏组织Wnt4、Wnt9b、GSK-3β、β-catenin蛋白表达量;采用实时荧光PCR法检测大鼠肾脏组织Wnt4 mRNA、Wnt9b mRNA、GSK-3β mRNA、β-catenin mRNA表达量.结果:与正常组比较,模型组及各药物组大鼠24 h尿蛋白、尿红细胞数量显著增加(P＜0.05),大鼠肾组织Wnt4、Wnt9b、β-catenin蛋白及Wnt4 mRNA、Wnt9b mRNA、GSK-3β mRNA、β-catenin mRNA相对表达量显著增加(P＜0.05),GSK-3β蛋白相对表达量降低(P＜0.05);与模型组比较,各药物组大鼠24 h尿蛋白、尿红细胞数量显著降低(P＜0.05),大鼠肾组织Wnt4、Wnt9b、GSK-3β、β-catenin蛋白及Wnt4 mRNA、Wnt9b mRNA、GSK-3β mRNA、β-catenin mRNA相对表达量显著降低(P＜0.05).结论:荆花消紫合剂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,其可能与Wnt/β-catenin信号通路的激活相关.
Abstract
Objective:To explore the protective effect of Jinghua Xiaozi mixture on purpuric nephritis in rats and the mechanism of action on Wnt/β-catenin signaling pathway.Methods:A total of 60 healthy male SD rats with SPF grade were selected.Totally 10 rats were randomly selected as the blank group,and the remaining 50 rats were established as experimental IgA nephropathy rat models by combined administration of BSA,LPS and CCl4.The successfully modeled IgA nephropathy rats were randomly divided into model group,Jinghua Xiaozii mixture(high,medium and low)dose group[(10.5 mL/(kg·d),7.0 mL/(kg·d),3.5 mL/(kg·d)],and Western medicine control group[piperazine ferulate tablets 13.5 mg/(kg·d)],10 rats in each group,continuous administration for 4 weeks.Urine analyzer was used to detect urine red blood cell count and 24-hour urine protein quantification in rats.Transmission electron microscopy was used to observe the lesions of kidney tissue in rats.Western blotting was used to detect the protein expressions of Wnt4,Wnt9b,GSK-3β and β-catenin in rat kidney tissues.Real-time fluorescence PCR were used to detect the expressions of Wnt4 mRNA,Wnt9b mRNA,GSK-3β mRNA and β-catenin mRNA in rat kidney tissues.Results:Compared with the blank group,the number of 24-hour urine protein and urine red blood cells were increased significantly in the model group and each drug group(P＜0.05),and the relative expression of Wnt4,Wnt9b,β-catenin protein and Wnt4 mRNA,Wnt9b mRNA,GSK-3β mRNA,β-catenin mRNA in kidney tissue were increased significantly(P＜0.05),while the relative expression of GSK-3β protein was decreased(P＜0.05).Compared with the model group,the number of 24-hour urine protein and urine red blood cells were decreased significantly in each drug group(P＜0.05),and the relative expression of Wnt4,Wnt9b,GSK-3β,β-catenin protein and Wnt4 mRNA,Wnt9b mRNA,GSK-3β mRNA,β-catenin mRNA in kidney tissue were significantly decreased(P＜0.05).Conclusion:Jinghua Xiaozi mixture can effectively reduce the urine red blood cell count and 24-hour urine protein quantification and alleviate kidney injury in rats with henoch-schonlein purpura nephritis,which may be related to the activation of Wnt/β-catenin signaling pathway.
维普
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