基于PERK/eIF2α通路探讨清热止血方联合雷公藤多苷治疗紫癜性肾炎大鼠的疗效机制  

Therapeutic mechanism of Qingre Zhixue Formula combined with tripterygium wilfordi polyglycosides in the treatment of Henoch-Schonlein purpura nephritis rats based on PERK/eIF2αpathway

作　　者：王龙[1,2] 丁樱[1,2] 徐闪闪[2] 张霞[2] 韩姗姗[1,2] WANG Long;DING Ying;XU Shanshan;ZHANG Xia;HAN Shanshan(School of Pediatrics,Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Affiliated Hospital of Henan University of CM,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,郑州450003

出　　处：《中华中医药杂志》2024年第7期3424-3429,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82305310,No.82305311);河南省自然科学基金项目(No.222300420488);河南省中医药科学研究专项课题(No.2023ZY2044);河南中医药大学第一附属医院博士启动金项目(No.2022BSJJ2005)。

摘　　要：目的:探讨清热止血方联合雷公藤多苷(GTW)改善紫癜性肾炎(HSPN)内皮损伤的机制。方法:72只大鼠随机分为空白对照组及造模组,造模组构建HSPN大鼠模型,将造模成功后的大鼠随机分为模型组、清热止血方组、GTW组、联合组、泼尼松组。其中空白对照组及模型组予0.9%氯化钠溶液灌胃,余治疗组予相应药物灌胃,2次/d,干预4周后,检测大鼠24 h尿总蛋白定量;光镜下观察大鼠肾组织PAS染色情况;荧光镜下观察大鼠肾组织肾小球IgA沉积情况;ELISA检测血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、血管细胞黏附因子-1(VCAM-1)、血管内皮生长因子(VEGF)表达;Western Blot检测大鼠肾组织磷酸化RNA蛋白激酶样内质网激酶(p-PERK)、磷酸化真核翻译起始因子2α(p-eIF2α)、葡萄糖调节蛋白78(GRP78)蛋白表达。结果:与空白对照组比较,模型组大鼠24 h尿总蛋白定量显著升高(P<0.05),模型组可见肾小球系膜增生,肾小球毛细血管管腔受压变狭窄,且伴有少量炎症细胞浸润,肾小球系膜区可见有团块状绿色荧光,且强度较高,血清IL-1β、TNF-α、VCAM-1、VEGF表达显著升高(P<0.05),p-PERK、p-eIF2α、GRP78蛋白表达显著升高(P<0.05)。与模型组比较,联合组治疗可显著降低24 h尿总蛋白定量水平(P<0.05),改善肾组织系膜增生及IgA荧光沉积,降低血清中IL-1β、TNF-α、VCAM-1、VEGF表达(P<0.05),降低p-PERK、p-eIF2α、GRP78蛋白表达(P<0.05)。结论:清热止血方联合GTW可有效降低HSPN模型大鼠尿总蛋白水平,减轻肾脏病理及内皮损伤,其机制可能与调控肾组织内质网应激PERK/eIF2α通路有关。Objective:To explore the mechanism of Qingre Zhixue Formula combined with tripterygium wilfordi glycosides(GTW)in improving endothelial damage in Henoch-Schonlein purpura nephritis(HSPN).Methods:A total of 72 rats were randomly divided into a blank control group and a modeling group.The modeling group was used to construct an HSPN rat model.After successful modeling,the rats were randomly divided into a model group,a Qingre Zhixue Formula group,a GTW group,a combination group,and a prednisone group.The blank control group and model group were given physiological saline by gavage,while the other treatment groups were given corresponding drugs by gavage twice a day.After 4 weeks of intervention,the 24-hour urine total protein quantification of rats was measured;the PAS staining of rat kidney tissue under light microscope was observed;the deposition of IgA in the glomerulus of rat kidney tissue under fluorescence microscopy was observed;ELISA for detecting serum expression of IL-1β,TNF-α,VCAM-1 and VEGF;the protein expression of p-PERK,p-eIF2α,GRP78 in rat kidney tissue were detected by Western Blot.Results:Compared with the blank control group,the total protein content in the urine in 24 h of the model group rats was significantly increased(P<0.05),in the model group,glomerular mesangial hyperplasia was observed,with narrowing of the glomerular capillary lumen due to compression,accompanied by a small amount of inflammatory cell infiltration,and there were clustered green fluorescence with high intensity in the glomerular mesangial area of rats,the expression of IL-1β,TNF-α,VCAM-1 and VEGF significantly increased(P<0.05),while the expression of p-PERK and p-eIF2α,GRP78 protein significantly increased(P<0.05).Compared with the model group,the combination group could significantly reduce the quantitative level of total urine protein in 24 h(P<0.05),improve renal mesangial hyperplasia and IgA fuorescence deposition,and reduce serum IL-1β,TNF-α,VCAM-1 and VEGF(P<0.05),reduce the expression of p-PERK and p-eIF2�

关 键 词：紫癜性肾炎 清热止血方 雷公藤多苷 RNA蛋白激酶样内质网激酶/真核翻译起始因子2α通路 动物模型 机制 内质网应激 炎症因子 

分 类 号：R285.5[医药卫生—中药学]