趋化因子2促进肝再生中脂肪的形成

doi:10.16098/j.issn.0529-1356.2016.05.022

摘要/Abstract

摘要：

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%。pEGFP-N1-CCL2转染组肝再生率、肝脏指数、ALT、AST和TBIL含量均高于pEGFP-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，p-MEK1/2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。

关键词: 趋化因子2, 脂肪合成, 肝再生, 苏丹Ⅳ, 实时定量聚合酶链反应, 免疫印迹法, 大鼠

Abstract:

Objective To elucidate the effect of chemokine 2 (CCL2) on liver regeneration and its mechanism. Methods Rats were randomly divided into 3 groups with 10 rats in each group. The plasmids were transferred into rats by the hydraulic gene transfer technology. Transfection efficiency was observed under a fluorescence microscope after 6 hours. The weight of regenerated liver was weighed, and the liver regenerating rate, together with the liver index were calculated to observe the liver regeneration. Liver function was observed by the content of alanine aminotransferase, aspartate aminotransferase and total bilirubin in serum. Sultan Ⅳ staining was used to observe the lipid accumulation, and Real-time PCR to detect the expression of genes related to lipid metabolism; The expression of phosphorylated mitogen extracellular kinase 1/2 (p-MEK1/2) and phosphorylated extra-cellular-regulated kinase l/2 (p-ERKl/2) were detected by Western blotting. Results After 6 hours, the positive rate of green fluorescent protein in each group was more than 30%. After transfection with pEGFP-N1-CCL2, liver regenerating rate, liver index, alanine amiotransferase(ALT), aspartate aminotransferase(AST) and total bilirubin(TBIL) were all higher than those of the pEGFP-N1 group. As the transgenic time was prolonged, the expression of the adipose related genes increased, more lipid drops appeared, and the expression of p-MEK1/2 and p-ERKl/2 increased. Conclusion Chemokine 2 may promote liver regeneration by increasing lipid synthesis via the MEK/ERK pathway.

Key words: Chemokine 2, Lipid synthesis, Liver regeneration, Sudan Ⅳ, Real-time PCR, Western blotting, Rat

邢雪琨* 武红艳林俊堂丰慧根原志庆. 趋化因子2促进肝再生中脂肪的形成[J]. 解剖学报, 2016, 47(5): 697-702.

XING Xue-kun* WU Hong-yan2LIN Jun-tang FENG Hui-gen YUAN Zhi-qing. Effect of chemokine 2 on the formation of lipid on liver regeneration[J]. AAS, 2016, 47(5): 697-702.

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