脂质诱导的巨噬细胞M1/M2型极化对肝细胞脂质代谢的影响  

Effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes

作　　者：王祺 许钦瑜 吴惠敏 华静[1] Wang Qi;Xu Qinyu;Wu Huimin;Hua Jing(Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200001, China)

机构地区：[1]上海交通大学医学院附属仁济医院消化内科 上海市消化疾病研究所,200001

出　　处：《中华肝脏病杂志》2018年第4期276-281,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金（81170374、81470842）

摘　　要：目的探讨脂质诱导的巨噬细胞M1／M2型极化对肝细胞脂质代谢的影响。方法分别以饱和脂肪酸——棕榈酸（PA）、单不饱和脂肪酸——油酸（OA）和多不饱和脂肪酸——二十二碳六烯酸（DHA）孵育巨噬细胞RAW264．7，并制备条件培养液（CM）。胶原酶原位灌注法分离C57BL／6小鼠肝细胞，体外巨噬细胞CM培养原代肝细胞。Real—time PCR检测巨噬细胞M1／M2型极化基因表达，以Real-time PCR和Western blot检测肝细胞脂质合成和分解代谢相关基因的mRNA和蛋白表达，油红O染色检测肝细胞内脂质沉积情况。多组样本数据间比较采用单因素方差分析，两两比较采用t检验。结果与对照组相比，PA诱导巨噬细胞M1型极化[肿瘤坏死因子α、白细胞介素（IL）-6mRNA表达水平显著升高，F≥22．68，P值均〈0．01]，OA诱导巨噬细胞M1／M2型混合极化[IL-6、甘露糖受体c2（Mrc2）和IL-10mRNA表达水平升高，F≥4．94，P值均〈0．05]，而DHA则诱导巨噬细胞M2型极化（Mrc2、IL-10mRNA表达水平增加，F≥4．94，P值均〈0．01）。CM—PA显著增加肝细胞脂质合成基因固醇调节元件结合蛋白1C（SREBP1C）、乙酰辅酶A羧化酶1（ACC1）mRNA表达（F≥5．66，P值均〈0．01）和脂肪酸合酶、ACCl蛋白表达（F≥38．34，P值均〈0．05），并降低脂质分解蛋白脂酰辅酶A氧化酶1（ACOX1）表达（F=154．48，P〈0．01），促进肝细胞内大量脂滴沉积；CM—OA影响肝细胞部分脂质代谢基因表达；而CM—DHA显著增加肝细胞脂质分解基因肉碱棕榈酰转移酶1A（CPT1A）mRNA表达（F=10．30，P〈0．01）和ACOX1、CPT1A蛋白表达（P值均〈0．05），并降低脂质合成蛋白SREBP1C、ACC1的表达（F≥65．84，P值均〈0．05），CM—DHA孵育的肝细胞内见少量脂滴沉积。结论不同脂肪酸通过诱导巨噬细胞M1／M2型极化改变影响肝细胞和肝脏脂质代谢平衡，从而促进或延缓Objective This study aims to explore the effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes. Methods RAW264.7 macrophages were incubated with different kinds of fatty acids including saturated fatty acids-palmitic acid （PA）, monounsaturated fatty acidsoleic acid （OA） and polyunsaturated fatty acids-docosahexaenoic acid （DHA）, and cell culture supernatants were collected to prepare conditioned medium （CM）. Hepatocytes were isolated by in situ petfusion of the liver with collagenase in mice, and a macrophage-hepatocyte CM co-culture system was established. Macrophage M1/ M2 phenotype markers were detected by Real-time PCR. Lipid synthesis and decomposition related mRNA and protein expressions in hepatocytes were detected by Real-time PCR and Western Blot. Lipid depositions in hepatocytes were detected by oil red O staining. An analysis of variance was used for comparison of means between multiple groups. Results Compared with control groups, PA polarized macrophages to a M1 phenotype （expression of TNF-α and IL-6 significantly increased, F≥ 22.68,P〈 0.01）, OA polarized macrophages to a M1/ M2 mixed phenotype （expression of IL-6, Mrc2 and 1L-10 increased F≥4.94, P 〈 0.05） and DHA polarized macrophages to a M2 phenotype （expression of Mrc2 and IL-10 significantly increased, F ≥ 4.94,P〈 0.01）. CMPA significantly increased lipid synthesis related genes, including SREBP 1 C, ACC 1 mRNA expression （F ≥ 5.66, P 〈 0.01） and FASN, ACC1 protein expression （F ≥ 38.34, P 〈 0.05） in hepatocytes, and decreased lipid decomposition gene ACOX1 protein expression （F=154.48, P 〈 0.01）. CM-OA affected several lipid metabolism genes expression. CM-DHA significantly increased CPT1A mRNA expression （F= 10.30,P 〈 0.01） and ACOX1, CPT1A protein expression （F ≥ 47.06,P 〈 0.05）, and decreased SREBP1C, ACC1 protein expression （F≥ 65.84, P 〈 0.05） in hepatocytes. Massive lipid droplets were deposited in hepatocytes in CM-P

关 键 词：脂肪肝 脂肪酸 巨噬细胞极化 脂质代谢 

分 类 号：R575.5[医药卫生—消化系统]