基础医学与临床 ›› 2007, Vol. 27 ›› Issue (2): 143-147.

• 研究论文 • 上一篇    下一篇

p-FAK(Tyr397)在大鼠肝纤维形成中的作用及机制研究

张晓岚 霍晓霞 申建刚 魏娟   

  1. 河北医科大学第二医院消化科 河北医科大学第二医院消化科 河北医科大学第二医院消化科 河北医科大学第二医院消化科
  • 收稿日期:2006-06-02 修回日期:2006-07-21 出版日期:2007-02-25 发布日期:2007-02-25
  • 通讯作者: 霍晓霞

The of phosphorylated fdcal adhesion kinase in rat hepatic fibogenesis and the study on mechanism

  

  • Received:2006-06-02 Revised:2006-07-21 Online:2007-02-25 Published:2007-02-25

摘要: 目的 探讨在肝纤维化发生中,黏着斑激酶(FAK)酪氨酸磷酸化与肝星状细胞(HSC)增殖的关系,并从细胞周期角度探讨其促HSC增殖的机制。方法 采用胆总管结扎(BDL)方法建立大鼠肝纤维化模型;免疫组织化学方法测定α-平滑肌肌动蛋白(α-SMA)表达;Western blot检测p-FAK(Tyr397)、细胞周期蛋白D1(cyclin D1)和细胞周期蛋白依赖激酶4(CDK4)在肝组织中的含量。结果 正常大鼠肝组织有少量α-SMA分布,随着肝纤维化发展,α-SMA阳性细胞明显增多;肝组织中p-FAK(Tyr397)、cyclin D1及CDK4蛋白表达逐渐增加,造模4周时达峰值。多元相关分析示α-SMA与p-FAK(Tyr397)正相关(r=0.964,P<0.01);α-SMA与cyclin D1、CDK4正相关(r=0.953,0.906;P<0.01);p-FAK(Tyr397)与cyclin D1、CDK4亦明显正相关(r=0.969,0.893;P<0.01)。结论 FAK磷酸化促HSC增殖及肝纤维化形成机制可能与调控细胞周期相关蛋白有关。

Abstract: Objective To observe the relationship between phosphorylation of focal adhesion kinase(FAK) and hepatic stellate cell(HSC) proliferation in fibrotic rat liver, and to study the mechanism of phosphorylated FAK regulating HSC proliferation from the cell cycle point of view. Methods The rat hepatic fibrosis was induced by bile duct ligation (BDL). Histopathological changes were evaluated by hematoxylin and eosin staining, and by Masson's trichrome method. Numbers of activated HSCs were quantified after alpha smooth muscle actin(α-SMA) staining. The proteins of p-FAK(Tyr397), Cyclin D1 and cyclin dependent kinase4 (CDK4) in the liver tissue were assayed by Western blot. Results ①With the development of hepatic fibrosis, the positive cells of α-SMA increased obviously. The positive areas of the rat livers in model groups at week 1 to 4 (12.88%±2.63%, 22.65%±2.16%, 27.45%±1.86%, 35.25%±2.34%) were larger than that in control group (5.88%±1.46%), P<0.01. ②Western blot analysis showed that in the sham-operated group, the p-FAK(Tyr397), Cyclin D1 and CDK4 was expressed a little. But with the development of liver fibrosis, the expression of the three kinds of proteins increased gradually in model groups at week 1 to 4. ③α-SMA correlated with p-FAK (Tyr397) positively, r value was 0.964, P<0.01; α-SMA correlated with Cyclin D1 and CDK4 positively, r value were 0.953 and 0.906 respectively, P<0.01; P-FAK(Tyr397) correlated with Cyclin D1 and CDK4 positively, r value were 0.969 and 0.893 respectively,P<0.01. Conclusions In hepatic fibrogenesis process, HSC became active and proliferated, while the expression of p-FAK (Tyr397), Cyclin D1 and CDK4 increased. P-FAK (Tyr397) correlated positively with Cyclin D1 and CDK4. The mechanism of phosphorylated FAK promoting hepatic fibrosis probably concerned with regulating cell cycle related protein.