SHP2表达变化对四氯化碳诱导的肝纤维化大鼠肝组织中Akt表达的影响  

Effects of SHP2 expression changes on Akt in liver tissues of rats with hepatic fibrosis induced by carbon tetrachloride

作　　者：郝礼森 王薇 季景秀 蒋美钰 苗笑佳 高莹莹 莫艳波 王静 HAO Lisen;WANG Wei;JI Jingxiu;JIANG Meiyu;MIAO Xiaojia;GAO Yingying;MO Yanbo;WANG Jing(Department of Gastroenterology,the Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,China;Department of Gastroenterology,Cangzhou Central Hospital,Cangzhou 061000,China)

机构地区：[1]华北理工大学附属医院消化内科,河北唐山063000 [2]沧州市中心医院消化内科,061000

出　　处：《国际消化病杂志》2024年第1期29-35,共7页International Journal of Digestive Diseases

基　　金：河北省自然科学基金面上项目(H2018209366)。

摘　　要：目的 探讨含SH2结构域的蛋白酪氨酸磷酸酶2(SHP2)过表达及低表达对四氯化碳(CCl4)诱导的肝纤维化大鼠肝组织中蛋白激酶B(Akt)的影响。方法选取160只健康雄性SD大鼠,随机分为对照组、模型组、AdGFP组、Ad-SHP2组和Ad-shRNA/SHP2组,每组32只。采用腹腔注射CCl4法构建大鼠肝纤维化模型,经大鼠尾静脉分别将表达绿色荧光蛋白(GFP)的空病毒Ad-GFP、表达野生型SHP2及GFP的腺病毒Ad-SHP2、表达GFP并携带靶向SHP2的短发夹RNA(shRNA)的腺病毒Ad-shRNA/SHP2注入大鼠体内。各组分别于造模第2、4、6、8周随机选取8只大鼠,留取肝组织标本。采用实时荧光定量PCR法检测各组大鼠肝组织中SHP2、Akt的mRNA表达水平,采用蛋白质印迹法检测各组大鼠肝组织中SHP2、Akt及磷酸化Akt(p-Akt)的蛋白表达水平,采用H-E染色法观察各组大鼠肝组织的病理变化,采用Masson三色染色法观察各组大鼠肝组织的胶原沉积情况。结果 靶向SHP2的shRNA及外源性野生型SHP2基因成功导入肝纤维化大鼠体内,并使大鼠肝组织中SHP2呈低表达或过表达。与模型组及Ad-GFP组比较,Ad-SHP2组大鼠的肝纤维化程度加重,而Ad-shRNA/SHP2组大鼠的肝纤维化程度则减轻。在同一造模时间点(第2、4、6、8周)对各组大鼠肝组织中Akt的m RNA和蛋白表达水平,以及p-Akt蛋白表达水平进行比较,结果显示Ad-GFP组、Ad-SHP2组、Ad-shRNA/SHP2组及模型组均显著高于对照组(P均<0.05),而各时间点的Ad-GFP组、Ad-SHP2组、Ad-shRNA/SHP2组及模型组的Akt表达水平差异均无统计学意义(P均>0.05);与模型组及Ad-GFP组大鼠肝组织中p-Akt表达水平相比较,AdshRNA/SHP2组在各时间点均显著降低(P均<0.05),Ad-SHP2组在各时间点均显著升高(P均<0.05),模型组与Ad-GFP组的p-Akt表达水平差异均无统计学意义(P均>0.05)。结论 在CCl4诱导的大鼠肝纤维化病程中,肝组织中SHP2过表达可通过促进Akt磷酸化增强Akt的活性,而肝Objective This paper intends to explore the effects of overexpression and low expression of SH2-containing protein tyrosine phosphatase 2(SHP2)on serine-threonine protein kinase(Akt)in liver tissue of carbon tetrachloride(CCl4)induced liver fibrosis rats.Methods One hundred and sixty healthy male SD rats were selected and randomly divided into the control group,the model group,the Ad-GFP group,the Ad-SHP2 group,and the Ad-shRNA/SHP2 group,with 32 rats in each group.A rat liver fibrosis model was constructed by intraperitoneal injection of CCl4.The empty virus Ad-GFP expressing green fluorescent protein(GFP),adenovirus Ad-SHP2 expressing wild-type SHP2 and GFP,and adenovirus Ad-shRNA/SHP2 carrying short hairpin RNA(shRNA)targeting SHP2 expressing GFP were injected into the tail vein of rats.Eight rats were randomly selected from each group at 2nd,4th,6th,and 8th week of modeling,and the liver tissue samples were collected.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression levels of SHP2 and Akt in the liver tissues of rats in each group.Western blotting was used to detect the protein expression levels of SHP2,Akt,and phosphorylated Akt(p-Akt)in the liver tissues of rats in each group.H-E staining was used to observe the pathological changes of liver tissue in each group.Masson trichrome staining was used to observe the collagen deposition in liver tissue in each group.Results SHP2-targeted shRNA and wild-type SHP2 gene are successfully introduced into rats with hepatic fibrosis,and the expression of SHP2 in liver tissues of rats is down-regulated and up-regulated.Compared with the model group and the Ad-GFP group,the degree of liver fibrosis in the Ad-SHP2 group aggravated,while the degree of liver fibrosis in the Ad-shRNA/SHP2 group alleviated.The mRNA and protein expression levels of Akt in the liver tissues of rats in each group are compared,as well as the expression levels of p-Akt protein at the same modeling time point(2nd,4th,6th,and 8th week).Those in the Ad-GFP group,th

关 键 词：含SH2结构域的蛋白酪氨酸磷酸酶2 肝纤维化 蛋白激酶B 

分 类 号：R575.2[医药卫生—消化系统]