益肾活血方对单侧输尿管梗阻小鼠肾间质纤维化的改善作用及机制研究  

Effect and Mechanism of Yishen Huoxue Recipe(益肾活血方)on Renal Interstitial Fibrosis in UUO Mice

作　　者：钟建[1] 罗芳[2] 方桂玉 卢登勇 石晓冬 向清[1] 王琳[1] 龚爱媚 熊冲 ZHONG Jian;LUO Fang;FANG Guiyu;LU Dengyong;SHI Xiaodong;XIANG Qing;WANG Lin;GONG Aimei;XIONG Chong(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China;Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530023 [2]广西中医药大学研究生学院,广西南宁530001

出　　处：《中华中医药学刊》2023年第7期15-20,I0011,I0012,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81760807,82060820)。

摘　　要：目的 探讨益肾活血方对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾间质纤维化(renal interstitial fibrosis,RIF)及能量代谢因子腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)、沉默信息调节因子3(silent information regulator 3,SIRT3)、肉毒碱棕榈酰基转移酶1A(carnitine palmitoyltransferase 1A,CPT1A)、过氧化物酶增殖激活受体-γ共激活子-1(peroxisome proliferator-activated receptor-γ coactivator-1α,PGC-1α)水平的影响。方法 90只C57BL/6雄性小鼠随机分为6组:假手术组,模型组,白藜芦醇组,益肾活血方低、中、高剂量组,每组15只。造模术后第1天(d1)开始,益肾活血方高、中、低剂量组分别按(34.4、17.2、8.6 g·kg^(-1))进行灌胃,白藜芦醇组予50 mg/(kg·d)灌胃,假手术组及模型组予等体积生理盐水灌胃,每日1次。造模成功后第7、14、21天(d7、d14、d21),每组随机处死小鼠5只,检测尿素氮(blood urea nitrogen,BUN)、血清肌酐(serum creatinine,Scr)的含量,HE及Masson染色观察肾组织病理结构变化,Western blot及qPCR法检测肾组织AMPK、SIRT3、CPT1A、PGC-1α蛋白表达及mRNA表达。结果 与假手术组相比,同时间段的模型组BUN、Scr水平显著升高(P<0.001)。与模型组相比,同时间段白藜芦醇组和益肾活血方(低、中、高剂量)组BUN、Scr浓度均明显下降(P<0.01或P<0.001);与白藜芦醇组相比,d21益肾活血方高剂量组对肾损伤治疗效果更明显(P<0.001)。模型组中AMPK、SIRT3、PGC-1α、CPT1A蛋白表达量及mRNA表达与假手术组相比明显下降(P<0.01或P<0.001),与模型组相比,白藜芦醇组、益肾活血方(中、高)剂量组d14与d21AMPK、SIRT3、CPT1A、PGC-1α蛋白表达及mRNA表达显著上调(P<0.001)。结论 益肾活血方能改善肾功能,延缓肾纤维化,这可能与激活线粒体上能量因子SIRT3及AMPK表达,促使PGC-1α、CPT1A活化有关。Objective To investigate the effects of Yishen Huoxue Recipe(益肾活血方)on renal interstitial fibrosis(RIF)and the influence on the levels of AMP-activated protein kinase(AMPK),silent information regulator 3(SIRT3),carnitine palmitoyltransferase 1A(CPT1A)and peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)energy metabolism factors in mice with unilateral ureteral obstruction(UUO).Methods Ninety male C57BL/6 mice were randomly divided into 6 groups:sham operation group,model group,resveratrol group,Yishen Huoxu Recipe low,medium and high dose groups,15 mice in each group.On the 1st day after modeling(d1),Yishen Huoxue Recipe high,medium and low dose groups were given intragastric administration of Yishen Huoxu Recipe(34.4,17.2 and 8.6 g·kg^(-1)).The resveratrol group was given resveratrol with the dose of 50 mg/(kg·d)by intragastric administration.The sham operation group and model group were given the equal volume of normal saline by intragastric administration,once a day.On the 7th,14th and 21st day(d7,d14,d21)after successful modeling,5 mice in each group were randomly sacrificed to detect the contents of urea nitrogen(BUN)and serum creatinine(Scr).Hematoxylin-eosin(HE)staining and Masson staining were used to observe the pathological structure changes of the renal tissue.The protein and mRNA expressions of AMPK,SIRT3,CPT1A and PGC-1αwere detected by Western blot and real-time quantitative qPCR.Results Compared with those of the sham operation group,the levels of BUN and Scr in the model group significantly increased at the same time(P<0.001).Compared with those of the model group,the concentrations of BUN and Scr in the resveratrol group and Yishen Huoxue Recipe low,medium and high dose groups significantly decreased(P<0.01 or P<0.001).Compared with those of the resveratrol group,the Yishen Huoxue Recipe high dose group had more obvious therapeutic effect on kidney injury on d21(P<0.001).The protein and mRNA expressions of AMPK,SIRT3,PGC-1αand CPT1A in the model group were signifi

关 键 词：益肾活血方 单侧输尿管梗阻小鼠 肾间质纤维化 AMPK SIRT3 CPT1A PGC-1Α 

分 类 号：R285.5[医药卫生—中药学]