[FRNK对肝星状细胞激活和迁移的影响]

[Effects of FRNK on activation and migration of hepatic stellate cells].

作者信息

Hu R H, Zhao X K, Huang T, Zou G L

机构信息

Department of Infectious Diseases, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China.

出版信息

Zhonghua Nei Ke Za Zhi. 2021 Apr 1;60(4):362-367. doi: 10.3760/cma.j.cn112138-20200625-00621.

DOI:10.3760/cma.j.cn112138-20200625-00621

PMID:33765707

Abstract

To investigate the effect of focal adhesion kinase related non kinase (FRNK) on the activation and migration of hepatic stellate cells (HSCs). 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 (CCl). 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 and α-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. 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 FRNKmice (153±13)% was more serious than that in WT (100%) group. The expression of PY397-FAK and α-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 and α-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 <0.01. 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 and inhibition of Rac and Rho activation.

摘要

探讨粘着斑激酶相关非激酶（FRNK）对肝星状细胞（HSCs）激活和迁移的影响。2019年3月至2019年9月，贵州医科大学附属医院将人肝组织分为健康对照组和纤维化组。将C57BL/6小鼠分为野生型（WT）和FRNK基因敲除型（FRNK）组。用四氯化碳（CCl）建立肝纤维化模型。之后，用腺病毒载体构建FRNK基因过表达载体（Ad-FRNK）。采用苏木精-伊红（HE）和Masson染色评估肝组织的病理变化和纤维沉积。用蛋白质免疫印迹法检测PY397-FAK和α-SMA蛋白的表达。提取小鼠原代HSCs，采用划痕实验检测FRNK对HSCs迁移的影响，用蛋白质免疫印迹法检测Rac和Rho的激活情况。肝纤维化患者肝组织中PY397-FAK蛋白表达明显高于健康对照组（0.88±0.09 vs. 0.73±0.09）。FRNK明显低于对照组（0.68±0.09 vs. 0.79±0.11）。建立动物模型后，FRNK基因敲除小鼠肝纤维化程度（153±13）%比WT组（100%）更严重。PY397-FAK和α-SMA蛋白表达明显升高（2.50±0.23 vs. 0.75±0.09，1.46±0.20 vs. 0.92±0.10）。FRNK基因重新表达后（100%），肝纤维化程度主要得到逆转[（74±6）%]，PY397-FAK和α-SMA的表达相应降低（0.68±0.11 vs. 1.12±0.19，0.68±0.10 vs. 0.85±0.06）。体外实验中，FRNK抑制HSCs的迁移[WT∶FRNK∶Ad-FRNK，（339±49）%∶（580±53）%∶（259±33）%]以及Rac和Rho蛋白的激活（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），差异均有统计学意义（均P<0.01）。FRNK可抑制促成肝纤维化的HSCs的激活和迁移。其潜在机制与下调PY397-FAK以及抑制Rac和Rho激活有关。