UGT2B28、FABP5、CYP2C9基因在脂肪酸代谢异常所致大鼠肝纤维化发生中的作用

肝脏 ›› 2022, Vol. 27 ›› Issue (9): 999-1003.

UGT2B28、FABP5、CYP2C9基因在脂肪酸代谢异常所致大鼠肝纤维化发生中的作用

宋敬茹, 武超, 曹红燕, 谢咚, 王铮, 刘璐, 王丹, 孙明瑜, 边艳琴

201203 上海中医药大学附属曙光医院肝病研究所(宋敬茹,武超,谢咚,王铮,刘璐,王丹,孙明瑜);上海中医药大学附属上海市中西医结合医院(曹红燕);上海中医药大学附属光华中西医结合医院(边艳琴)

收稿日期:2021-11-27 出版日期:2022-09-30 发布日期:2022-10-27
通讯作者: 边艳琴,Email:xiaobian504@126.com
基金资助:
国家中医药管理局第四批中医优才(2017-124);光华育人工程培育计划(2020-M-1);上海中医药大学后备中医卓越人才计划(2020);上海市科委专项(19401972300);国家中医药管理局中医肝胆病重点学科、慢性肝病虚损重点研究室和上海市中医临床重点实验室资助项目(20DZ2272200)。

Important roles of UGT2B28, FABP5, and CYP2C9 in the development of rat liver fibrosis induced by abnormal fatty acid metabolism

SONG Jing-ru, WU Chao, CAO Hong-yan, XIE Dong, WANG Zheng, LIU Lu, WANG Dan, SUN Ming-yu, BIAN Yan-qin

Shuguang Hospital, Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Shanghai Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai, 200082, China; Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai 200052, China

Received:2021-11-27 Online:2022-09-30 Published:2022-10-27
Contact: BIAN Yan-qin,Email:xiaobian504@126.com

摘要/Abstract

摘要： 目的脂肪酸代谢异常是二甲基亚硝胺(Dimethylniyrosamine,DMN)诱导导致大鼠肝纤维化发生的重要机制之一,本研究进一步阐明DMN诱发肝纤维化发生的关键调控基因。方法通过复制经典的DMN诱导肝纤维化大鼠模型,在造模2周、4周时分别留取大鼠血清及肝脏组织。血清进行肝功能及血脂分析,肝组织进行全基因芯片分析,借助生物信息分析软件,对芯片数据进行深度挖掘。结果造模2周、造模4周模型组大鼠血清ALT分别为(45.83 ± 5.03)U/L、(88.06±15.51)U/L高于对照组(25.28 ± 3.03)U/L;血清AST分别为(69.17 ± 5.20)、(108.10 ± 17.31)U/L,高于对照组(61.44 ± 11.82)U/L;血清TG分别为(1.11 ± 0.30)、(0.77 ± 0.10)mmol/L,高于对照组(0.77 ± 0.22)mmol/L;血清FFA分别为(0.26 ± 0.04)、(0.49 ± 0.18)mmol/L,高于对照组(0.51 ± 0.18)mmol/L,差异均有统计学意义(均P<0.05)。55个共享差异基因(Log Ratio >2)主要参与了肝纤维化发生的9条信号通路,主要涉及细胞增殖、脂肪酸代谢、细胞凋亡、细胞死亡等生物学功能。共享差异基因UGT2B28在2周模型组和4周模型组中明显上调大于5倍,FABP5在2周模型组中明显上调大于5倍,CYP2C9在4周模型组中明显下调大于5倍。共享差异基因UGT2B28、FABP5和CYP2C9三个基因均参与了脂代谢的所有过程。结论共享差异基因UGT2B28、FABP5和CYP2C9在脂肪酸代谢异常引起的肝纤维发生机制中具有关键作用。

关键词: 肝纤维化, 脂肪酸代谢, 生物信息分析, 二甲基亚硝胺

Abstract: Objective Purpose Abnormal fatty acid metabolism has been found to be one of the mechanisms of dimethylnitrosamine (DMN)-induced hepatic fibrosis. In this study, we further elucidated the key regulatory genes of fatty acid metabolism in DMN-induced rat model of hepatic fibrosis. Methods The classical rat model of DMN-induced liver fibrosis was replicated. Serum and liver tissues were collected at 2 and 4 weeks of modeling. The serum samples were analyzed for liver function and lipid metabolism. Whole gene microarray was performed with the liver tissues and the microarray data were deeply mined with bioinformatic analysis software. Results Serum ALT and AST levels were (25.28 ± 3.03)U/L and (69.17 ± 5.20)U/L, and (45.83 ± 5.03)U/L and (108.10 ± 17.31)U/L in the model group at 2 weeks and 4 weeks of modeling, respectively, which was significantly higher than those of (25.28 ± 3.03)U/L and (61.44 ± 11.82)U/L in the control group (P<0.05); Similarly, serum TG and FFA levels were (1.11 ± 0.30) and (0.26 ± 0.04), and (0.77 ± 0.10) and (0.49 ± 0.18) mmol/L in the model group at 2 weeks and 4 weeks of modeling, respectively, which was significantly higher than (0.77 ± 0.22) and (0.51 ± 0.18) mmol/L in the control group (P<0.01 for TG, and P<0.05 for FFA). There were 55 shared differential genes (Log Ratio >2) were identified, which were enriched in 9 signaling pathways associated with cell proliferation, fatty acid metabolism, apoptosis, cell death and other biological functions. The shared differential gene UGT2B28 was significantly up-regulated (>5-fold) in 2 weeks and 4 weeks liver samples of the model group, while FABP5 in the liver samples of the model group at 2 weeks and CYP2C9 in the liver samples of the model group at 4 weeks were significantly up-regulated (>5-fold). UGT2B28, FABP5 and CYP2C9 were involved in all processes of lipid metabolism. Conclusion The shared differential genes UGT2B28, FABP5 and CYP2C9 play key roles in the mechanism of liver fibrogenesis associated with abnormal fatty acid metabolism.

Key words: Hepatic fibrosis, Fatty acid metabolism, Bioinformatic analysis, Dimethylnitrosamine

宋敬茹, 武超, 曹红燕, 谢咚, 王铮, 刘璐, 王丹, 孙明瑜, 边艳琴. UGT2B28、FABP5、CYP2C9基因在脂肪酸代谢异常所致大鼠肝纤维化发生中的作用[J]. 肝脏, 2022, 27(9): 999-1003.

SONG Jing-ru, WU Chao, CAO Hong-yan, XIE Dong, WANG Zheng, LIU Lu, WANG Dan, SUN Ming-yu, BIAN Yan-qin. Important roles of UGT2B28, FABP5, and CYP2C9 in the development of rat liver fibrosis induced by abnormal fatty acid metabolism[J]. Chinese Hepatolgy, 2022, 27(9): 999-1003.

参考文献

[1] 邝胜利, 胡兵. 肝纤维化大鼠模型研究进展. 实验动物与比较医学, 2008(01):62-66.
[2] 边艳琴, 曹红燕, 李建缘, 等. 脂肪酸代谢异常是二甲基亚硝胺诱导的大鼠肝纤维化发病机制之一. 世界科学技术-中医药现代化, 2016, 18(02):241-249.
[3] Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev, 2017, 121:27-42.
[4] Ala-Kokko L, Pihlajaniemi T, Myers JC, et al. Gene expression of type I, III and IV collagens in hepatic fibrosis induced by dimethylnitrosamine in the rat. Biochem J, 1987, 244(1):75-79.
[5] 边艳琴. 茵陈蒿汤及其组分复方抗肝纤维化的效应机制研究. 上海中医药大学, 2012.
[6] Zhang Z, Sun ZZ, Xiao X, et al. Mechanism of BDE209-induced impaired glucose homeostasis based on gene microarray analysis of adult rat liver. Arch Toxicol, 2013, 87: 1557-1567.
[7] 刘佳萱, 李彬彬, 张利芬, 等. 脂质代谢和肝纤维化关系的研究进展. 第二军医大学学报, 2018, 39(05):531-534.
[8] Molenaar MR, Vaandrager AB, Helms JB. Some lipid droplets are more equal than others: different metabolic lipid droplet pools in hepatic stellate cells. Lipid Insights, 2017, 10:1178635317747281.
[9] 边艳琴, 刘平, 孙明瑜. 基于方证相关理论解析肝硬化湿热内蕴病机. 世界科学技术-中医药现代化, 2016, 18(09):1477-1482.
[10] Senda A, Mukai Y, Hayakawa T, et al. Angiotensin II receptor blockers inhibit the generation of epoxyeicosatrienoic acid from arachidonic acid in recombinant CYP2C9, CYP2J2 and human liver microsomes. Basic Clin Pharmacol Toxicol, 2017, 121: 239-245.
[11] Louet M, Labbé C M, Fagnen C, et al. Insights into molecular mechanisms of drug metabolism dysfunction of human CYP2C9*30. PloS one, 2018, 13(5):e0197249.
[12] Ohnishi A, Murakami S, Akizuki S, et al. In vivo metabolic activity of CYP2C19 and CYP3A in relation to CYP2C19 genetic polymorphism in chronic liver disease. J Clin Pharmacol, 2005, 45: 1221-1229.
[13] Frye RF, Zgheib NK, Matzke GR, et al. Liver disease selectively modulates cytochrome P450--mediated metabolism. Clin Pharmacol Ther, 2006, 80: 235-245.
[14] Iso T, Maeda K, Hanaoka H, et al. Capillary endothelial fatty acid binding proteins 4 and 5 play a critical role in fatty acid uptake in heart and skeletal muscle. Arterioscler Thromb Vasc Biol, 2013, 33(11):2549-2557.
[15] Thumser AE, Moore JB, Plant NJ. Fatty acid binding proteins: tissue-specific functions in health and disease. Curr Opin Clin Nutr Metab Care, 2014, 17(2):124-129.
[16] Ohata T, Yokoo H, Kamiyama T, et al. Fatty acid-binding protein 5 function in hepatocellular carcinoma through induction of epithelial-mesenchymal transition. Cancer Med, 2017, 6(5):1049-1061.
[17] Jeong CY, Hah YS, Im Cho B, et al. Fatty acid-binding protein 5 promotes cell proliferation and invasion in human intrahepatic cholangiocarcinoma. Oncol Rep, 2012, 28(4):1283-1292.
[18] Dallaglio K, Marconi A, Truzzi F, et al. E-FABP induces differentiation in normal human keratinocytes and modulates the differentiation process in psoriatic keratinocytes in vitro. Exp Dermatol, 2013, 22(4):255-261.
[19] Song J, Zhang H, Wang Z, et al. The role of FABP5 in radiation-induced human skin fibrosis. Radiat Res, 2018, 189(2):177-186.
[20] Belledant A, Hovington H, Garcia L, et al. The UGT2B28 sex-steroid inactivation pathway is a regulator of steroidogenesis and modifies the risk of prostate cancer progression. Eur Urol, 2016, 69(4):601-609.
[21] Le PH, Kuo CJ, Hsieh YC, et al. Ages of hepatocellular carcinoma occurrence and life expectancy are associated with a UGT2B28 genomic variation. BMC cancer, 2019, 19: 1190.