Hck、Lyn和SSeCKS在Kupffer细胞中的表达及意义

The significance and expression of Hck,Lyn and SSe CKS in Kupffer cells

目的 :探讨Src家族相关激酶(Hck、Lyn)和Src抑制的蛋白激酶C底物(Src-suppressed C kinase substrate,SSe CKS)在Kupffer细胞中的表达及意义。方法:应用Real-time PCR的方法检测正常Kupffer细胞和炎症Kupffer细胞中Hck、Lyn和SSe CKS的表达水平;用Western Blot的方法检测Hck、Lyn和SSe CKS在Kupffer细胞中的表达水平。构建非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)大鼠模型,采用免疫组织化学(immunohistochemistry,IHC)方法检测Hck、Lyn和SSe CKS在大鼠肝脏组织Kupffer细胞中的表达水平。结果:RT-PCR显示Hck、Lyn和SSe CKS在小鼠Kupffer细胞中均有表达,用60%CCl4损伤液刺激后Hck、Lyn表达明显升高(P<0.01),SSe CKS表达减少,但差异无统计学意义(P>0.05);Western Blot结果显示在Kupffer细胞中Hck、SSe CKS表达较为明显,Lyn未见明显表达;IHC结果显示Hck、Lyn和SSe CKS在大鼠正常肝组织Kupffer中均有表达;在炎症组织中Hck、Lyn的表达明显增加,而SSe CKS的表达却显著减少。在炎症刺激后Hck、Lyn的表达与Kupffer细胞数呈正相关,而SSe CKS的表达与Kupffer细胞数呈负相关。结论:Hck、Lyn和SSe CKS参与了NASH的发生发展,可作为NASH炎症反应的潜在调控靶点;检测其表达有助于NASH炎症程度的判断,为临床上治疗NASH提供一定的客观依据。

Objective：To analyze the significance and expression of Src family related kinases （Hck, Lyn）/SSeCKS in Kupffer cells. Methods： Real-time PCR was used to detect the expression of Hck,Lyn and SSeCKS in both normal and inflammation Kupffer cells. The expression of Hck, Lyn and SSeCKS were detected by Western Blot. The normal SD rats were treated with high-fat for 8 weeks and the models were established. The expression of Hck、Lyn and SSeCKS in kupffers cells were expressed by immunohistochemistry（IHC）. Results：RT-PCR showed that Hck, Lyn and SSeCKS were all expressed in Kupffer cells in mice. The expression of Hck, Lyn increased significantly （P〈0.05） after stimulating with 60% CCl4 liquid damage, the expression of SSeCKS were decreased, but without statistical significance（P〉0.05）;Western Blot showed that Hck, Lyn and SSeCKS were all expressed in Kupffer cells, especially the expressoin of Hck and SSeCKS was obvious and it has no obvious expression of Lyn in rat Kupffer cells. IHC showed that Hck, Lyn and SSeCKS were all expressed in normal liver tissue Kupffer cells in rats. In the expression of inflammatory tissue Hck and Lyn increased significantly, but the SSeCKS expression is significantly reduced. The expression of Hck and Lyn were positively correlated with number of Kupffer cells, while the expression of SSeCKS was negatively correlated with number of Kupffer cells after the inflammation stimulation. Conclusion：Hck, Lyn and SSeCKS are involved in the occurrence and development of NASH, that can be used as a potential target of NASH inflammation. The study can help clinical diagnosis of NASH inflammation degree, provide some objective basis for clinical treatment of NASH.