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目的探讨角蛋白18(keratin18,K18)在33和52位丝氨酸(Ser)磷酸化水平变化的作用及其与肝纤维化的关系。方法 6周龄Balb/C小鼠30只,分为3组(对照组和2个实验组),每组10只。对照组腹腔内注射橄榄油;实验组腹腔内注射四氯化碳(CCl4)和橄榄油的混合液(1∶9),10 ml/kg,每周2次,连续4周,分别在2周和4周处死小鼠。应用组织化学免疫荧光法检测正常对照小鼠和肝纤维化小鼠肝组织中K18及其磷酸化Ser33和Ser52的表达及其相对亚细胞定位;免疫印迹法(Western blotting)检测正常对照小鼠和肝纤维化小鼠肝组织的K18及其磷酸化Ser33和Ser52水平。结果组织化学免疫荧光结果显示,K18在正常对照小鼠和肝纤维化小鼠肝组织中均有表达,但在小鼠肝纤维化不同阶段无明显差异;在正常对照小鼠肝组织中表达比较弱,而随着肝纤维化的进展表达增强。West-ern blotting结果,肝纤维化小鼠肝组织中的Ser33和Ser52磷酸化的K18表达水平显著增加,尤其是Ser33表达增加更为明显。
Objective To investigate the role of keratin 18 (K18) in serine (Ser) phosphorylation at 33 and 52 and its relationship with hepatic fibrosis. Methods Thirty Balb / C mice of 6 weeks old were divided into 3 groups (control group and 2 experimental groups), 10 in each group. The control group was intraperitoneally injected with olive oil. The experimental group was intraperitoneally injected with a mixture of CCl4 and olive oil (1: 9) at 10 ml / kg twice a week for 4 weeks, And mice were sacrificed 4 weeks later. The expression and relative subcellular localization of K18, phosphorylated Ser33 and Ser52 in the liver tissue of normal control and hepatic fibrosis mice were detected by histochemical immunofluorescence. The expression of K18, K18 and its phosphorylated Ser33 and Ser52 levels in hepatic tissue of liver fibrosis mice. Results The results of histochemical immunofluorescence showed that K18 was expressed in the liver of normal mice and hepatic fibrosis mice, but no significant difference was observed in the different stages of liver fibrosis in mice. Compared with normal control mice, Weak, but with the progress of liver fibrosis increased expression. West-ern blotting results showed that phosphorylation of K18 at Ser33 and Ser52 in hepatic tissue of liver fibrosis mice increased significantly, especially Ser33 expression increased more significantly.