黏着斑激酶相关非激酶对肝星状细胞活化及迁移功能的影响  被引量:2

Effects of FRNK on activation and migration of hepatic stellate cells

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作  者:胡睿涵 赵雪珂 黄韬 邹高亮 Hu Ruihan;Zhao Xueke;Huang Tao;Zou Gaoliang(Department of Infectious Diseases,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区:[1]贵州医科大学附属医院感染科,贵阳550004

出  处:《中华内科杂志》2021年第4期362-367,共6页Chinese Journal of Internal Medicine

基  金:国家自然科学基金(81860115)。

摘  要:目的探讨黏着斑激酶相关非激酶(FRNK)对肝星状细胞(HSCs)活化及迁移功能的影响。方法2019年3—9月,收集贵州医科大学附属医院肝胆外科9例肝纤维化患者肝组织标本。人体肝组织分为健康对照组与纤维化组。C57BL/6实验小鼠分为野生型(WT)、FRNK基因敲除型(FRNK-/-)两组,以四氯化碳(CCl4)进行肝纤维化造模,完成后使用腺病毒载体构建FRNK基因过表达型(Ad-FRNK)。通过HE、Masson染色观察肝组织病理改变,Western蛋白印迹法检测磷酸化黏着斑激酶(PY397-FAK)与α平滑肌肌动蛋白(α-SMA)蛋白表达;提取小鼠原代HSCs,细胞划痕实验检测FRNK对HSCs迁移功能的影响,及对Rac、Rho活化的影响。结果肝纤维化人体肝组织PY397-FAK蛋白表达量显著高于健康对照组(0.88±0.09比0.73±0.09),FRNK低于健康对照组(0.68±0.09比0.79±0.11),P值均<0.01。CCl4造模后,FRNK-/-组小鼠肝纤维化[(153±13)%]较WT组(100%)程度更明显,PY397-FAK与α-SMA蛋白表达量高于WT组(2.50±0.23比0.75±0.09,1.46±0.20比0.92±0.10),P值均<0.01。在FRNK-/-组小鼠体内重新导入FRNK基因(100%)后,小鼠肝纤维化程度减轻[(74±6)%],PY397-FAK与α-SMA蛋白表达量减少(0.68±0.11比1.12±0.19,0.68±0.10比0.85±0.06),P值均<0.01。体外实验显示,FRNK可抑制HSCs的迁移功能[WT︰FRNK-/-︰Ad-FRNK为(339±49)%︰(580±53)%︰(259%±33)%],并抑制Rac和Rho蛋白的活化(Rac为0.54±0.07比0.91±0.10比0.77±0.12,Rho为0.45±0.05比0.64±0.06比0.53±0.07),P值均<0.01。结论FRNK可通过抑制HSCs的活化及迁移功能改善肝纤维化,其机制可能与下调PY397-FAK、抑制Rac和Rho的活化有关。Objective To investigate the effect of focal adhesion kinase related non kinase(FRNK)on the activation and migration of hepatic stellate cells(HSCs).Methods Human liver tissue was divided into healthy control group and fibrosis group from March 2019 to September 2019 in Affiliated Hospital of Guizhou Medical University.C57BL/6 mice were divided into wild type(WT)and FRNK gene knockout type(FRNK-/-)groups.The liver fibrosis model was established with carbon tetrachloride(CCl4).After that,FRNK gene overexpression(Ad-FRNK)was constructed with adenovirus vector.HE and Masson staining were used to evaluate the pathological changes and fiber deposition of liver tissue.Western blot was used to detect the expression of PY397-FAK andα-SMA protein.Mouse primary HSCs were extracted,and the effect of FRNK on HSCs migration was detected by wound healing,activation of Rac and Rho was detected by Western blot.Results The expression of PY397-FAK protein in human liver tissue with hepatic fibrosis was significantly higher than that in healthy control group(0.88±0.09 vs.0.73±0.09).FRNK was significantly lower than that in control group(0.68±0.09 vs.0.79±0.11).After animal model was set up,the degree of liver fibrosis in FRNK-/-mice(153±13)%was more serious than that in WT(100%)group.The expression of PY397-FAK andα-SMA protein was significantly elevated(2.50±0.23 vs.0.75±0.09,1.46±0.20 vs.0.92±0.10).After FRNK gene was re-expressed(100%),the degree of liver fibrosis was mainly reversed[(74±6)%],and the expression of PY397-FAK andα-SMA was accordingly decreased(0.68±0.11 vs.1.12±0.19,0.68±0.10 vs.0.85±0.06).In vitro,FRNK inhibited the migration of HSCs[WT∶FRNK-/-∶Ad-FRNK,(339±49)%∶(580±53)%∶(259±33)%]and the activation of Rac and Rho proteins(Rac:0.54±0.07 vs.0.91±0.10 vs.0.77±0.12,Rho:0.45±0.05 vs.0.64±0.06 vs.0.53±0.07),all P<0.01.Conclusions FRNK can inhibit the activation and migration of HSCs which contributed to liver fibrosis.The potential mechanism is related to down regulation of PY397-FAK

关 键 词:肝硬化 黏着斑蛋白酪氨酸激酶类 细胞迁移分析 

分 类 号:R575.2[医药卫生—消化系统]

 

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