PICK1在肝纤维化中的表达变化及功能研究

发布时间：2018-08-25 17:41

【摘要】：目的研究蛋白激酶C结合蛋白1(PICK1)在小鼠肝纤维及活化的肝星状细胞中的表达变化,并探讨其对人肝星状细胞株(LX-2)活化的影响。方法建立四氯化碳(CCl4)诱导的小鼠肝纤维化模型,观察PICK1在肝纤维化过程中的表达变化;转化生长因子β1(TGF-β1)刺激LX-2活化,Western blot测定PICK1的蛋白表达情况;转染PICK1过表达质粒至LX-2中,再以TGF-β1诱导活化,Western blot检测PICK1、α平滑肌肌动蛋白(α-SMA)、I型胶原(Col1a1)和Smad2、3及其磷酸化水平的蛋白表达情况。结果正常组小鼠肝脏中PICK1表达较高,随着肝纤维化的加重,PICK1的表达逐渐递减。TGF-β1诱导活化的LX-2细胞中PICK1表达降低。转染PICK1过表达质粒后,TGF-β1诱导的α-SMA、Colla1的表达明显减少,且Smad2、3磷酸化水平显著下降。结论 PICK1在纤维化肝组织及活化的HSC中表达下调。过表达PICK1可抑制TGF-β1诱导的LX-2活化,可能是通过抑制TGF-β/Smad通路而发挥作用,为肝纤维化的防治研究提供了新的思路和靶点。
[Abstract]:Objective to investigate the expression of protein kinase C binding protein 1 (PICK1) in mouse hepatic fibers and activated hepatic stellate cells (HSCs) and to investigate the effect of protein kinase C binding protein 1 (PICK1) on the activation of human hepatic stellate cell line (LX-2). Methods the liver fibrosis model of mice induced by carbon tetrachloride (CCl4) was established to observe the expression of PICK1 during hepatic fibrosis, the expression of PICK1 protein was detected by LX-2 activation blot stimulated by transforming growth factor 尾 1 (TGF- 尾 1), and the overexpression plasmid of PICK1 was transfected into LX-2. The expression of PICK1, 伪 smooth muscle actin (伪 -SMA) type I collagen (Col1a1) and Smad2,3 and their phosphorylation levels were detected by TGF- 尾 1-induced activation Western blot. Results the expression of PICK1 in the liver of normal mice was higher, and the expression of PICK1 decreased gradually with the exacerbation of hepatic fibrosis. The expression of PICK1 in activated LX-2 cells induced by TGF- 尾 1 decreased. The expression of 伪 -SMA-Colla1 induced by TGF- 尾 1 was significantly decreased and the phosphorylation level of Smad2,3 was significantly decreased after transfection of PICK1 overexpression plasmid. Conclusion the expression of PICK1 is down-regulated in fibrotic liver tissue and activated HSC. Overexpression of PICK1 can inhibit the LX-2 activation induced by TGF- 尾 1, which may play a role by inhibiting the TGF- 尾 / Smad pathway, which provides a new idea and target for the prevention and treatment of hepatic fibrosis.
【作者单位】：安徽医科大学药学院;安徽医科大学肝病研究所;安徽医科大学安徽省创新药物产业共性研究院;
【基金】：国家自然科学基金(编号:81273526、81473268) 安徽省重点科技攻关项目(编号:1301042212) 安徽省自然科学基金(编号:1308085MH145) 高等学校博士学科点专项科研基金(编号:20123420120001)
【分类号】：R575.2

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