CD226基因敲除对小鼠肝纤维化的影响  

Effect of CD226 gene knockout on liver fibrosis in mice

作　　者：刘辉 杨涛 汪帅 王栋 阴继凯 Liu Hui;Yang Tao;Wang Shuai;Wang Dong;Yin Jikai(Graduate School of Air Force Medical University,Xian 710032,China;Department of General Surgery,the Second Afiliated Hospital of Air Force Medical University,Xian 710038,China)

机构地区：[1]空军军医大学研究生院,西安710032 [2]空军军医大学第二附属医院普通外科,西安710038

出　　处：《中华肝胆外科杂志》2024年第10期776-781,共6页Chinese Journal of Hepatobiliary Surgery

基　　金：空军军医大学第二附属医院重点项目(2019LCYJ005)。

摘　　要：目的探究CD226基因敲除(CD226^(-/-))对四氯化碳(CCl 4)诱导小鼠肝纤维化模型的影响。方法选取雄性野生型C57BL/6小鼠和CD226^(-/-)C57BL/6小鼠各8只,6~8周龄,体重分别为(22.16±2.24)g和(21.84±2.50)g。从野生型和CD226^(-/-)小鼠中各取4只予以腹腔注射CCl 4建立肝纤维化模型,即肝纤维化组和CD226^(-/-)肝纤维化组,剩余的各4只分别作为对照组和CD226^(-/-)对照组。观察CD226^(-/-)对肝质量、肝脏指数及Ishak纤维化评分的影响。HE、Masson及天狼猩红染色观察肝脏损伤和肝纤维化情况。免疫组织化学、蛋白质印迹法检测肝脏组织I型胶原(Collagen I)、α-平滑肌肌动蛋白(α-SMA)蛋白表达水平。流式细胞术分析外周血和肝脏T淋巴细胞比例及其胞内IFN-γ分泌水平。实时荧光定量PCR(qPCR)检测小鼠肝脏组织中转化生长因子-β1(TGF-β1)mRNA的表达水平。结果与肝纤维化组相比,CD226^(-/-)肝纤维化组小鼠的肝质量[(2.00±0.14)g比(1.41±0.16)g]及肝脏指数(7.25±0.59比5.33±0.30)均增加,Ishak纤维化评分[(6.75±0.96)分比(10.00±1.41)分]降低,差异均具有统计学意义(均P<0.05)。HE染色结果显示,肝纤维化组和CD226^(-/-)肝纤维化组小鼠的肝脏组织均显示肝组织结构紊乱,肝细胞变性坏死,但CD226^(-/-)肝纤维化组小鼠的炎症细胞募集程度轻于肝纤维化组。天狼猩红、Masson染色发现,与肝纤维化组相比,CD226^(-/-)肝纤维化组小鼠胶原纤维沉积减少。免疫组化染色结果显示,与肝纤维化组相比,CD226^(-/-)肝纤维化组小鼠肝脏组织中Collagen I[(4.38±0.51)%比(6.55±1.40)%]和α-SMA[(0.77±0.20)%比(1.20±0.24)%]的表达均减少,差异均具有统计学意义(均P<0.05)。蛋白质印迹法检测结果显示,与肝纤维化组相比,CD226^(-/-)肝纤维化组小鼠肝组织中Collagen I蛋白的相对表达降低(0.35±0.14比1.66±0.73),差异具有统计学意义(P<0.05)。流式细胞术结果显示,与肝纤维化组相比,CD226ObjectiveTo investigate the effect of CD226 gene knockout(CD226^(-/-))on carbon tetrachloride(CCl 4)-induced liver fibrosis in mice.MethodsEight 6-8 week-old male wild-type and CD226^(-/-)C57BL/6 mice were used,with weights of(22.16±2.24)g and(21.84±2.50)g,respectively.Four mice each from wild-type and CD226^(-/-)mice were injected intraperitoneally with CCl 4 to establish a liver fibrosis model,namely,the liver fibrosis group and the CD226^(-/-)liver fibrosis group,and the remaining four mice each corresponded to the control group and the CD226^(-/-)control group.The effect of CD226^(-/-)on liver mass,liver index and Ishak fibrosis score were observed.HE,Masson,and Sirius red staining were performed to observe the liver injury and liver fibrosis.Immunohistochemistry and Western blot were used to detect the protein expression levels of collagen type I(Collagen I)andα-smooth muscle actin(α-SMA)in liver tissues.Flow cytometry was used to analyze the proportion of T lymphocytes and their intracellular IFN-γsecretion levels in the peripheral blood and liver tissues.The mRNA expression level of transforming growth factor-β1(TGF-β1)in the liver tissues was detected by real-time fluorescence quantitative PCR(qPCR).ResultsCompared to the liver fibrosis group,the CD226^(-/-)liver fibrosis group showed an increase in liver mass[(2.00±0.14)g vs.(1.41±0.16)g]and liver index(7.25±0.59 vs.5.33±0.30),and a decrease in Ishak's fibrosis score[(6.75±0.96)score vs.(10.00±1.41)score],and the differences were statistically significant(all P<0.05).HE staining showed disorganization of the liver tissue structure,and degeneration and necrosis of the hepatocytes both occurred in the liver fibrosis group and CD226^(-/-)liver fibrosis group,but the degree of inflammatory cell recruitment was milder in the CD226^(-/-)liver fibrosis group than the liver fibrosis group.Sirius red and Masson staining revealed reduced collagen fibre deposition in the CD226^(-/-)liver fibrosis group compared to the liver fibrosis group.Immunohistoch

关 键 词：肝纤维化 CD226 肝损伤 T淋巴细胞 四氯化碳 

分 类 号：R575.2[医药卫生—消化系统]