慢肾康宁对肾间质纤维化大鼠内质网应激PERK-ATF4通路的影响  

Effect of Manshen Kangning Formula on PERK/ATF4 Pathway in Endoplasmic Reticulum Stress in Renal Interstitial Fibrosis Rats

作　　者：孙升云[1] 金鑫[1] 黄少君[2] 徐梅[2] 颜显欣[2] 朱诗平[1] 何玲[2] 聂玲辉[2] 

机构地区：[1]暨南大学附属第一医院 [2]暨南大学医学院

出　　处：《中医杂志》2013年第3期236-240,共5页Journal of Traditional Chinese Medicine

基　　金：暨南大学科研培育与创新基金资助项目

摘　　要：目的探讨慢肾康宁对肾间质纤维化大鼠肾脏组织的保护作用及机制。方法 60只雄性Wistar大鼠,随机选取7只为正常组,其余53只采用2.5%腺嘌呤悬浮液灌胃建立肾间质纤维化模型,21d后将50只造模成功的大鼠随机分为模型组、氯沙坦组[10mg/(kg·d)]、慢肾康宁低剂量组[7.5g/(kg·d)]、慢肾康宁中剂量组[15g/(kg·d)]、慢肾康宁高剂量组[30g/(kg·d)],每组10只,分别给予相应药物灌胃,正常组和模型组以等量蒸馏水灌胃,连续30d。实验结束后,检测各组大鼠血清肌酐(SCr)、尿素氮(BUN)、24小时尿蛋白定量(24hMTP),估算肾小球滤过率(eGFR),HE和Masson染色观察肾组织病理变化,免疫组织化学方法检测肾组织内质网应激标志性蛋白葡萄糖调节蛋白78(GRP78),TUNEL法检测肾小管上皮细胞凋亡水平,实时定量PCR测定肾组织GRP78、转录激活因子4(ATF4)、CCAAT增强子结合蛋白同源蛋白(CHOP)mRNA表达水平。结果与模型组比较,氯沙坦组和慢肾康宁各剂量组SCr、BUN、24hMTP均显著下降(P<0.01),eGFR上升(P<0.01)。病理观察,模型组可见肾小管上皮细胞凋亡、坏死、脱落,肾小管扩张、炎性细胞浸润,肾间质内腺嘌呤代谢产物沉积和胶原纤维增生,氯沙坦组和慢肾康宁各组较模型组病理变化轻。与模型组比较,各给药组GRP78表达水平及细胞凋亡水平均下降(P<0.05或P<0.01)。各给药组GRP78、ATF-4、CHOPmRNA表达水平较模型组降低(P<0.05)。结论慢肾康宁对肾间质纤维化大鼠肾脏组织内质网应激PERK-eIF2α-ATF4-CHOP通路过度表达有很好的抑制作用,具有肾脏保护功能,这可能是它减轻肾间质纤维化的重要机制之一。Objective To observe the effect and mechanism of Manshen Kangning Formula （formula for chronic kidney disease, MKF） on endoplasmic reticulum stress in renal tissue of renal interstitial fibrosis rats. Methods Sixty male Wistar rats were Selected, seven wererandomly set into normal group, and other 53 were given 2.5% adenine to make renal interstitial fi- brosis model. After modeling, fifty successful model rats were randomized into model group, losartan group, MKF small-dose group, MKF middle-dose group and MKF large-dose group, with 10 in each. The treatment groups were given losartan sus- pension [10mg/（kg · d）], small-dose MKF suspension [7.5g/（kg · d）], middle-dose MKF suspension [15g/（kg· d）] and large-dose MKF suspension [30g/（kg · d）]respectively and the same amount of distilled water was given to the normal group and model group intragastrically. The administration lasted for 30 days. The serum creatinine （SCr）, blood urea nitrogen （BUN）, 24-hour urinary protein （24hUP） and estimated glomerular filtration rate （eGFR） were detected. The renal his- topathologieal changes were observed with HE and Masson staining. The glucose-regulated protein 78 （GRP78） was detected with immunohistochemical staining and the apoptotic cells were detected with TUNEL staining. Real Time PCR was used to detect the expression levels of GRP78 mRNA, activating tranSCription factor 4 （ATF4） mRNA and CCAAT/enhancer-binding protein homologous protein （CHOP） mRNA. Results Comparing with the model group, the levels of SCr, BUN and 24hUP were significantly decreased （P〈0.01）, but the eGFRs were significantly increased in the losartan group and MKF groups （P 〈0. 01）. Apoptosis, necrosis and shedding in renal tubular epithelial cells, tubular dilatation, inflammatory cell infiltration, adenine metabolite deposition and collagen fibers in renal interstitial tissue could be seen under the microscope in the model group. Comparing with the model group, the pathological changes we

关 键 词：慢肾康宁 肾间质纤维化 内质网应激 葡萄糖调节蛋白78 转录激活因子4 CCAAT增强子结合蛋白同源蛋白 

分 类 号：R285[医药卫生—中药学]