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目的探讨趋化因子2(CCL2)对肝再生的影响以及作用机制。方法大鼠随机分为3组,每组10只。液压转基因技术将质粒转入大鼠体内,6 h后荧光显微镜下观察转染效率。称量再生肝重量,计算肝再生率和肝脏指数以观察肝脏再生情况。测量血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)与总胆红素(TBIL)的含量以评估肝脏功能情况。苏丹Ⅳ染色观测脂肪聚积情况。Real-time PCR检测脂肪代谢相关基因的表达。Western blotting检测磷酸化丝裂原细胞外激酶1/2(p-MEK1/2)和磷酸化细胞外信号调节激酶l/2(p-ERKl/2)的表达情况。结果转质粒后6 h各组绿色荧光蛋白表达量均大于30%。p EGFP-N1-CCL2转染组肝再生率、肝脏指数、ALT、AST和TBIL含量均高于p EGFP-N1组。随转基因时间延长,脂肪代谢相关基因表达量增加,有较多猩红色脂肪滴出现,pMEK1/2和p-ERKl/2表达量增多。结论趋化因子2可能通过MEK/ERK通路增加脂肪合成,促进肝脏再生。
Objective To investigate the effect of chemokine 2 (CCL2) on liver regeneration and its mechanism. Methods The rats were randomly divided into three groups of 10 rats. The plasmids were transferred into rats by hydraulic gene transfer technology and the transfection efficiency was observed under a fluorescence microscope 6 hours later. Weighing regenerated liver weights, calculating liver regeneration rate and liver index to observe liver regeneration. The levels of serum ALT, AST and TBIL were measured to assess liver function. Sudan Ⅳ staining observed fat accumulation situation. Real-time PCR was used to detect the expression of fat metabolism related genes. Western blotting was used to detect the expression of p-MEK1 / 2 and p-ERK1 / 2 in phosphorylated mitogen. Results The expression of GFP in each group was more than 30% 6 h after transfection with plasmid. p EGFP-N1-CCL2 transfection group liver regeneration rate, liver index, ALT, AST and TBIL levels were higher than the p EGFP-N1 group. With the extension of GMO, the gene expression of fat metabolism increased, more scarlet lipid droplets appeared and the expression of pMEK1 / 2 and p-ERK1 / 2 increased. Conclusions Chemokine 2 may increase fat synthesis through MEK / ERK pathway and promote liver regeneration.