三七总皂苷对慢性肾衰竭大鼠AngⅡ/AT1R/ERK1/2信号通路的影响  

Effects of Panax notoginseng Saponins on the AngⅡ/AT1R/ERK1/2 Signaling Pathway of Rats with Chronic Renal Failure

作　　者：徐珂 黄学宽[1] 沈清[2] 罗洪玉 田珈瑜 邹波 左玲 王慕南 杨琴 侯科名 XU Ke;HUANG Xuekuan;SHEN Qing;LUO Hongyu;TIAN Jiayu;ZOU Bo;ZUO Ling;WANG Munan;YANG Qin;HOU Keming(College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China;Department of Nephrology,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院,重庆400016 [2]重庆医科大学附属第一医院,重庆400016

出　　处：《中药新药与临床药理》2021年第10期1417-1424,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：重庆市卫生和计划生育委员会、重庆市科学技术委员会联合资助项目(ZY201802149);重庆市技术创新与应用发展专项面上项目(cstc2019jscx-msxmX0119)。

摘　　要：目的研究三七总皂苷(PNS)对腺嘌呤诱导的慢性肾衰竭(CRF)大鼠AngⅡ/AT1R/ERK1/2信号通路的影响,探讨其延缓慢性肾衰竭进展的作用机制。方法将65只雄性SD大鼠随机分为正常组10只、造模组55只,正常组常规饲养,造模组采用250 mg·kg^(-1)·d^(-1)腺嘌呤混悬液连续灌胃28 d复制CRF大鼠模型。将造模成功大鼠随机分为模型组、贝那普利组(10 mg·kg^(-1))及PNS低、中、高(40、80、160 mg·kg^(-1))剂量组,灌胃给药,每日1次,连续28 d。检测大鼠24 h尿蛋白(24 h U-pro)及血清肌酐(Scr)、尿素氮(BUN)的含量;采用HE染色法观察肾组织病理形态,Masson染色法观察肾间质纤维化程度;采用ELISA法检测血清及肾组织匀浆中血管紧张素Ⅱ(AngⅡ)、血清中胱抑素C(Cys-C)的含量;采用Western Blot法分析肾组织中血管紧张素Ⅱ受体Ⅰ型(AT1R)、细胞外调节激酶(ERK1/2)、磷酸化ERK1/2(p-ERK1/2)蛋白表达情况;Real-time PCR法检测肾组织中AngⅡ、AT1R、结缔组织生长因子(CTGF)mRNA的表达;免疫组化法检测肾组织中AT1R蛋白的表达。结果与正常组比较,模型组大鼠的24 h U-pro、Scr、BUN、Cys-C水平显著升高(P<0.01),血清、肾组织匀浆中的AngⅡ含量明显增加(P<0.01),肾组织中的AngⅡ、AT1R、CTGF mRNA表达均明显上调(P<0.01),AT1R、p-ERK1/2蛋白表达均明显上调(P<0.01)。与模型组比较,给药组大鼠的Scr、BUN、24 h U-pro及Cys-C水平均明显降低(P<0.05,P<0.01),血清、肾组织中的AngⅡ含量均明显降低(P<0.05,P<0.01),肾组织中的AngⅡ、AT1R、CTGF mRNA表达均明显下调(P<0.05,P<0.01),AT1R、p-ERK1/2蛋白表达均明显下调(P<0.05,P<0.01)。结论三七总皂苷可改善CRF模型大鼠的肾功能和肾脏的病理学改变,其机制可能与调控AngⅡ/AT1R/ERK1/2信号通路有关。Objective To study the effects of Panax notoginseng saponins(PNS)on the AngⅡ/AT1R/ERK1/2 signaling pathway of rats with chronic renal failure(CRF),and to explore its mechanism of delaying the development of chronic renal failure.Methods 65 male Sprague-Dawley rats were randomly divided into normal group(n=10)and modeling group(n=55).The normal group was routinely reared,while the modeling group were administered by gavage with 250 mg·kg^(-1) adenine suspension for 28 days to replicate the rat model of CRF.After the model was successfully established,the survival model rats were randomly divided into model group,Benazepril(10 mg·kg^(-1))group and PNS(40,80,160 mg·kg^(-1))groups.Drug intervention lasted for 28 days.After the experiment,the 24-hour urine protein(24 h U-pro)was measured.The contents of serum creatinine(Scr)and blood urea nitrogen(BUN)in rats’serum were measured,the histological morphology was observed by HE staining,and the degree of renal interstitial fibrosis was observed by Masson staining.Enzyme linked immunosorbent assay(ELISA)was used to determine the contents of angiotensinⅡ(AngⅡ)in serum and renal homogenate and cystatin C(Cys-C)in serum.The protein levels of angiotensinⅡreceptor 1 subtype(AT1R),extracellular regulatory protein kinase 1/2(ERK1/2)and its phosphorylation were detected by Western Blot.The expression of AT1R protein in renal tissues were detected by immunohistochemistry.The expression of AngⅡ,AT1R,connective tissue growth factor(CTGF)mRNA in the kidney tissues were observed by the method of Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Results Compared with the normal group,the levels of 24 h U-pro,Scr,BUN and Cys-C in the model group rats were significantly increased(P<0.01),the contents of AngⅡin serum and kidney tissues were significantly increased(P<0.01),the expression of AngⅡ,AT1R,CTGF mRNA and AT1R,p-ERK1/2 protein in renal tissues were significantly increased(P<0.01).Compared with the model group rats,after the interventio

关 键 词：三七总皂苷 慢性肾衰竭 肾功能 AngⅡ/AT1R/ERK1/2信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]