芍药色素下调miR-21并激活SIRT1/FOXO1同路进而保护CCl_(4)诱导的大鼠急性肝损伤  

The protective effect of peonidin through SIR1/FOXO1 pathway via miR-21 on CCl_(4)-induced acute liver injury in rats

作　　者：侯延丽[1,2] 王鹏[2] 王娅宁[2] HOU Yan-li;WANG Peng;WANG Ya-ning(Medical School of Yan′an University,Yan′an Shaanxi 716000,China;Xi′an Innovation College of Yan′an University,Xi′an Shaanxi 710100,China)

机构地区：[1]延安大学医学院,陕西延安716000 [2]延安大学西安创新学院,陕西西安710100

出　　处：《毒理学杂志》2021年第5期405-411,共7页Journal of Toxicology

摘　　要：目的考察芍药色素是否通过微小RNA-21(miR-21)调控沉默信息调节因子1(SIRT1)/叉头框转录因子O1(FOXO1)通路保护四氯化碳(CCl_(4))诱导的大鼠急性肝损伤。方法将72只SD大鼠分为正常组、模型组(CCl_(4)处理)、芍药色素组(芍药色素处理)、芍药色素+miR-21组(芍药色素+miR-21处理)、EX527组(SIRT1特异性抑制剂EX527处理)和芍药色素+EX527组(芍药色素+EX527处理),每组12只。大鼠在第3、6和9天分别灌胃给予0.1、0.2和0.3 ml的CCl_(4),第10~16天灌胃给予芍药色素或静脉注射miR-21激动剂、腹腔注射EX527处理,1次/d。称重法检测大鼠的肝脏指数,全自动生化分析仪检测大鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,TUNEL染色检测各组大鼠肝细胞凋亡,酶联免疫吸附测定法(ELISA)检测大鼠肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)水平,免疫印迹实验(Western blot)分析大鼠肝组织中Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)、SIRT1、FOXO1和磷酸化FOXO1(p-FOXO1)表达水平,实时荧光定量PCR(qRT-PCR)测定大鼠肝组织中miR-21表达水平。结果与正常组比较,模型组大鼠肝脏指数、ALT、AST、MDA水平、肝细胞凋亡数量、Bax蛋白、miR-21的表达水平和FOXO1/p-FOXO1值升高(P<0.05),SOD水平、Bcl-2、SIRT1、FOXO1和p-FOXO1蛋白的表达水平降低(P<0.05),大鼠急性肝损伤造模成功。与模型组比较,芍药色素组大鼠的肝脏指数、ALT、AST、MDA水平、肝细胞凋亡数量、Bax蛋白、miR-21的表达水平和FOXO1/p-FOXO1值降低(P<0.05),SOD水平、Bcl-2、SIRT1、FOXO1和p-FOXO1蛋白的表达水平升高(P<0.05)。与芍药色素组比较,芍药色素+miR-21组大鼠肝组织的SIRT1、FOXO1和p-FOXO1蛋白水平降低(P<0.05),FOXO1/p-FOXO1值升高(P<0.05)。EX527组大鼠SIRT1、FOXO1和p-FOXO1蛋白水平降低(P<0.05),FOXO1/p-FOXO1水平升高(P<0.05)。而芍药色素+miR-21组或芍药色素+EX527组大鼠的肝脏指数、ALT、AST、MDA水平Objective To investigate whether peonidin regulates the silencing information regulator 1(SIRT1)/forkhead box transcription factor O1(FOXO1)pathway through microRNA-21(miR-21)to protect rats acute liver injury induced by carbon tetrachloride(CCl_(4)).Methods Seventy-two SD rats were divided into normal group,model group(CCl_(4) treatment),peonidin group(peonidin treatment),peonidin+miR-21 group(peonidin+miR-21 treatment),EX527 group(SIRT1 specific inhibitor EX527 treatment)and peonidin+EX527 group(peony pigment+EX527 treatment).A total of 12 rats in each group were intragastrically given 0.1 ml,0.2 ml,and 0.3 ml of CCl_(4) on the 3rd,6th,and 9th days,respectively,and were treated with peonidin or miR-21 agonist and EX527 on the 10th-16th day once a day.The liver index of rats was determined by weighing method,the automatic biochemical analyzer was applied to detect the levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),TUNEL staining was used to detecte the apoptosis of rat liver cells in each group,and the enzyme-linked immunosorbent assay(ELISA)was performed to measure levels of superoxide orgotein dismutase(SOD)and malondialdehyde(MDA)in rat liver tissues.We used Western blot to analyze expression levels of Bcl-2 related X protein(Bax),B cell lymphoma/leukemia-2(Bcl-2)SIRT1,FOXO1 and phosphorylated FOXO1(p-FOXO1).Real-time fluorescence quantitative PCR(qRT-PCR)was used to determine the expression level of miR-21 in rat liver tissues.Results Compared with the control group,the liver index,levels of ALT,AST and MDA,the number of hepatocyte apoptosis,the expression levels of Bax protein and miR-21 and FOXO1/p-FOXO1 value in model group increased(P<0.05),levels of SOD,protein expression levels of Bcl-2,SIRT1,FOXO1 and p-FOXO1 decreased(P<0.05);protein levels of SIRT1,FOXO1,p-FOXO1 in EX527 group decreased and FOXO1/p-FOXO1 levels increased(P<0.05).Compared with the model group,the liver index,levels of ALT,AST,MDA,the number of hepatocyte apoptosis,the expression levels of Bax protein

关 键 词：芍药色素 肝损伤 四氯化碳 MIR-21 SIRT1/FOXO1通路 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]