熊果酸对肾脏缺血再灌注损伤的保护作用及其机制  

Protective effect of ursolic acid on kidney ischemia-reperfusion injury and mechanism

作　　者：汤井源 韩鹏[2] 左文仁 苏学林 沈炀 俞秋 苏健[1] 朱清毅[1] Tang Jingyuan;Han Peng;Zuo Wenren;Su Xuelin;Shen Yang;Yu Qiu;Su Jian;Zhu Qingyi(Department of Urology,Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Department of Urology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]江苏省中医院南京中医药大学附属医院泌尿外科,210029 [2]江苏省人民医院泌尿外科,南京210029

出　　处：《中华实验外科杂志》2020年第8期1421-1424,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年科学基金项目(81902570);江苏省中医院创新发展基金专项(Y2018CX63)。

摘　　要：目的探讨熊果酸(UA)在肾脏缺血再灌注损伤(IRI)中的作用及其机制。方法30只C57BL/6J小鼠(购自南京大学模式动物研究所)随机分为3组,假手术组(Sham组)、肾脏缺血再灌注损伤组(IRI组)和UA预处理组(UA+IRI组)。采用右肾切除左肾蒂钳夹30 min方式构建小鼠肾脏IRI模型,检测各组血清肌酐(SCr)、尿素氮(BUN)变化,并利用糖原染色(PAS)观察肾脏组织形态学改变。同时检测各组肾脏组织内丙二醛(MDA)、羰基化蛋白(CP)含量以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,利用蛋白印迹法(Western blot)检测肾脏组织中核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)和醌氧化还原酶1(NQO1)蛋白表达水平。此外,应用HK-2细胞缺氧/复氧模型(H/R)进一步从细胞水平评估UA在IRI中的作用。两样本均数间比较采用t检验,多样本均数间比较采用单因素方差分析。结果相较于I/R组,UA+I/R组血清SCr[(0.61±0.09)mg/dl比(1.62±0.21)mg/dl]和BUN[(61.53±8.46)mg/dl比(135.38±15.61)mg/dl]值显著降低,肾小管损伤(1.55±0.14比3.52±0.23)明显减轻(t=7.316,P<0.05)。此外,UA预处理可降低小鼠肾脏内MDA[(4.13±0.71)nmol/mg比(7.65±0.89)nmol/mg]和CP含量[(2.92±0.33)nmol/mg比(5.36±0.48)nmol/mg],增加SOD[(45.48±4.16)U/mg比(22.15±3.36)U/mg]和CAT活性[(38.66±4.52)U/mg比(21.54±3.85)U/mg]。Western blot结果提示UA+IRI组Nrf2、HO-1和NQO1表达水平较IRI组明显上升。此外,UA预处理显著降低了H/R导致的细胞死亡增加和MDA含量上升[(1.14±0.12)nmol/ml比(2.15±0.15)nmol/ml],同时增加了细胞SOD活性[(25.86±1.85)U/ml比(13.74±1.46)U/ml,t=5.413,P<0.05],但在Nrf2稳定敲低表达的HK-2细胞中,UA并不能明显降低MAD含量和增加SOD活性。结论UA可以诱导Nrf2和下游基因HO-1和NQO-1的表达,提高机体抗氧化应激能力,从而保护肾脏IRI。Objective To investigate the role of ursolic acid(UA)in kidney ischemia-reperfusion injury(IRI)and its potential mechanism.Methods Thirty C57BL/6J mice(purchased from Model Animal Research Center of Nanjing University)were randomly divided into three groups:sham operation group(Sham group),kidney ischemia-reperfusion injury group(IRI group)and UA pretreatment group(UA+IRI group).The kidney IRI model was constructed by removing the right kidney and clamping the left kidney pedicle for 30 min.The levels of serum creatinine(SCr)and blood urea nitrogen(BUN)were determined in each group.The morphological changes of kidney tissue were observed by PAS staining.Simultaneously,the contents of malondialdehyde(MDA),carbonylated protein(CP),superoxide dismutase(SOD)and catalase(CAT)in kidney tissues were measured.Western blotting was used to detect the expression of nuclear factor erythoid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1)and NAD(P)H:Quinone oxidoreductase 1(NQO1)in kidney tissue.In addition,HK-2 cell hypoxia/reoxygenation model(H/R)was used to further evaluate the role of UA in IRI at the cellular level.The t-test was used to compare the two sample means,and the one-way analysis of variance was used to compare the multi-sample means.Results The levels of SCr[(0.61±0.09)vs.(1.62±0.21)mg/dl]and BUN[(61.53±8.46)vs.(135.38±15.61)mg/dl]and the degree of tubular damage(1.55±0.14 vs.3.52±0.23)were significantly lower in the UA+I/R group than those in the I/R group(t=7.316,P<0.05).In addition,UA pretreatment reduced MDA[(4.13±0.71)vs.(7.65±0.89)nmol/mg]and CP[(2.92±0.33)vs.(5.36±0.48)nmol/mg]levels and increased SOD[(45.48±4.16)vs.(22.15±3.36)U/mg]and CAT[(38.66±4.52)vs.(21.54±3.85)U/mg]activities in the kidney.Western blotting results showed that the expression levels of Nrf2,HO-1 and NQO1 in UA+IRI group were significantly higher than those in IRI group.In addition,UA pretreatment significantly reduced the increased cell death and MDA[(1.14±0.12)vs.(2.15±0.15)μmol/L]content induced by H/R,while incr

关 键 词：熊果酸 肾脏缺血再灌注损伤 氧化应激 核因子E2相关因子2 

分 类 号：R692[医药卫生—泌尿科学]