趋化因子2促进肝再生中脂肪的形成

邢雪琨* 武红艳 林俊堂 丰慧根 原志庆

doi:10.16098/j.issn.0529-1356.2016.05.022

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解剖学报 ›› 2016, Vol. 47 ›› Issue (5) : 697-702. DOI: 10.16098/j.issn.0529-1356.2016.05.022

生物工程学

趋化因子2促进肝再生中脂肪的形成

邢雪琨^1* 武红艳² 林俊堂¹ 丰慧根¹ 原志庆¹

作者信息 +

Effect of chemokine 2 on the formation of lipid on liver regeneration

XING Xue-kun ^1* WU Hong-yan²LIN Jun-tang¹ FENG Hui-gen¹ YUAN Zhi-qing¹

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2016.05.022

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]. Acta Anatomica Sinica. 2016, 47(5): 697-702 https://doi.org/10.16098/j.issn.0529-1356.2016.05.022

参考文献

［1］Jiang WW, Hao XX, Guo YQ, et al. Caspase signaling pathway regulates hepatocyte apoptosis in rat liver regeneration ［J］.Acta Anatomica Sinica,2012, 43(3): 359-365. (in Chinese)

蒋文文, 郝晓霞, 郭雅琼, 等. Caspase信号通路调控大鼠再生肝肝细胞的凋亡［J］.解剖学报,2012, 43(3): 359-365.

［2］Michalopoulos GK. Liver regeneration after partial hepatectomy: critical analysis of mechanistic dilemmas ［J］.Am J Pathol, 2010, 176(1): 2-13.

［3］Hemsworth J, Kumwenda C, Arimond M, et al. Lipid-based nutrient supplements increase energy and macronutrient intakes from complementary food among malawian infants ［J］.J Nutr, 2016, 146(2): 326-334.

［4］Bechmann LP, Hannivoort RA, Gerken G, et al. The interaction of hepatic lipid and glucose metabolism in liver diseases ［J］. J Hepatol, 2012, 56(4): 952-964.

［5］Glende EA, Jr Morgan WS. Alteration in liver lipid and lipid fatty acid composition after partial hepatectomy in the rat ［J］.Exp Mol Pathol, 1968, 8(2): 190-200.

［6］Delahunty TJ, Rubinstein D. Accumulation and release of triglycerides by rat liver following partial hepatectomy ［J］.J Lipid Res, 1970, 11(6): 536-543.

［7］Girard A, Roheim PS, Eder HA. Lipoprotein synthesis and fatty acid mobilization in rats after partial hepatectomy ［J］.Biochim Biophys Acta, 1971, 248(1): 105-113.

［8］Gove CD, Hems DA. Fatty acid synthesis in the regenerating liver of the rat ［J］.Biochem J, 1978, 170(1): 1-8.

［9］Walldorf J, Hillebrand C, Aurich H, et al. Propranolol impairs liver regeneration after partial hepatectomy in C57Bl/6-mice by transient attenuation of hepatic lipid accumulation and increased apoptosis ［J］. Scand J Gastroenterol, 2010, 45(4): 468-476.

［10］Newberry EP, Kennedy SM, Xie Y, et al. Altered hepatic triglyceride content after partial hepatectomy without impaired liver regeneration in multiple murine genetic models ［J］.Hepatology, 2008, 48(4): 1097-1105.

［11］Iwanaga N, Nakamura S, Oshima K, et al. Macrolides promote CCL2mediated macrophage recruitment and clearance of nasopharyngeal pneumococcal colonization in mice ［J］. J Infect Dis, 2015, 212(7): 1150-1159.

［12］O’Connor T, Borsig L, Heikenwalder M. CCL2-CCR2 Signaling in Disease Pathogenesis ［J］.Endocr Metab Immune Disord Drug Targets, 2015, 15(2): 105-118.

［13］Xu CSh, Xing XK, Yang XG, et al. Hydrodynamics-based transgene directively into rat regenerating liver in vivo ［J］.Acta Anatomica Sinica,2009, 40(4): 599-603. (in Chinese)

徐存拴, 邢雪琨, 杨献光, 等. 液压转基因技术应用于大鼠再生肝转基因实验［J］.解剖学报,2009, 40(4): 599-603.

［14］Antonelli A, Ferri C, Fallahi P, et al. CXCL10 and CCL2 chemokine serum levels in patients with hepatitis C associated with autoimmune thyroiditis ［J］.J Interferon Cytokine Res, 2009, 29(6): 345-351.

［15］Sarma NJ, Tiriveedhi V, Crippin JS, et al. Hepatitis C virus-induced changes in microRNA 107 (miRNA-107) and miRNA-449a modulate CCL2 by targeting the interleukin-6 receptor complex in hepatitis ［J］.J Virol, 2014, 88(7): 3733-3743.

［16］Affo S, Sancho-Bru P. CCL2: a link between hepatic inflammation, fibrosis and angiogenesis ［J］? Gut, 2014, 63(12): 1834-1835.

［17］Lin ZY, Chuang YH, Chuang WL. Cancer-associated fibroblasts up-regulate CCL2, CCL26, IL6 and LOXL2 genes related to promotion of cancer progression in hepatocellular carcinoma cells［J］.Biomed Pharmacother, 2012, 66(7):525-529.

［18］Murray AB, Strecker W, Silz S. Ultrastructural changes in rat hepatocytes after partial hepatectomy, and comparison with biochemical results ［J］.J Cell Sci,1981, 50:433-448.

［19］Strzyz P. Lipid metabolism: Lipid droplet growth in brown fat ［J］.Nat Rev Mol Cell Biol, 2016, 17(1): 3.

［20］Ohsaki Y, Cheng J, Fujita A, et al. Cytoplasmic lipid droplets are sites of convergence of proteasomal and autophagic degradation of apolipoprotein B ［J］.Mol Biol Cell, 2006, 17(6): 2674-2683.

［21］Goodman JM. The gregarious lipid droplet ［J］.J Biol Chem, 2008, 283(42): 28005-28009.
［22］Clement S, Juge-Aubry C, Sgroi A, et al. Monocyte chemoattractant protein-1 secreted by adipose tissue induces direct lipid accumulation in hepatocytes ［J］.Hepatology, 2008, 48(3): 799-807.
［23］Srinivasan SR, Chow CK, Glauert HP. Effect of the peroxisome proliferator ciprofibrate on hepatic DNA synthesis and hepatic composition following partial hepatectomy in rats ［J］.Toxicology, 1990, 62(3): 321-332.
［24］Shteyer E, Liao Y, Muglia LJ, et al. Disruption of hepatic adipogenesis is associated with impaired liver regeneration in mice ［J］.Hepatology, 2004, 40(6): 1322-1332.
［25］Ben-David H, Aruna BV, Seger R, et al. A 50-kDa ERK-like protein is up-regulated by a dual altered peptide ligand that suppresses myasthenia gravis-associated responses ［J］.Proc Natl Acad Sci USA, 2006, 103(48): 18232-18237.
［26］Dygai AM, Zhdanov VV, Miroshnichenko LA, et al. Role of PI3K, MAPK/ERK1/2, and p38 in implementation of the proliferative and differentiation potential of erythroid progenitors after blood loss ［J］.Bull Exp Biol Med, 2015, 158(4): 417-420.
［27］Fuhrman B, Nitzan O, Karry R, et al. Urokinase plasminogen activator (uPA) stimulates cholesterol biosynthesis in macrophages through activation of SREBP-1 in a PI3-kinase and MEK-dependent manner ［J］.Atherosclerosis, 2007, 195(2): e108-116.