棕榈酸通过肝细胞氧化应激反应激活炎症小体  

Palmitic acid induces hepatocellular oxidative stress and activation of inflammasomes

作　　者：许雯[1] 郭予斌[1] 李旭[1] 何美蓉[1] 刘思德[1] 

机构地区：[1]南方医科大学南方医院消化科,广东广州510515

出　　处：《南方医科大学学报》2016年第5期655-659,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81470844);广州市临床医学研究与转化中心(胃肠道早期肿瘤)试点建设项目(741569196402)~~

摘　　要：目的观察棕榈酸(PA)对肝细胞氧化应激反应及炎症小体产生的影响。方法(1)将正常小鼠肝细胞AML12分别用不同浓度的棕榈酸(0、0.15、0.25、0.4 mmol/L)处理;(2)将AML12细胞分为空白对照组(control组)、棕榈酸组(PA组)及棕榈酸+N-乙酰半胱氨酸组(PA+NAC组)并予以相应药物处理。24 h后检测细胞中脂肪沉积情况、细胞总ROS、线粒体来源ROS、NOX4蛋白表达、NOX4的定位以及炎症小体和IL-1β的表达。结果与control组相比,PA组细胞质中脂滴增加,细胞总ROS(12463.09±2.72 vs 6691.23±2.45,P=0.00)及线粒体来源ROS含量(64.98±0.94 vs 45.04±0.92,P=0.00)上升,NOX4、NLRP3、ASC、caspase-1蛋白表达增加,IL-1β表达增加(1603.52±1.32 vs 2629.33±2.57,P=0.00),且发现线粒体和NOX4在细胞质中存在共定位,而抗氧化剂NAC除了能使PA诱导的ROS产生减少(7782.15±2.87 vs 5445.6±1.17,P=0.00),还可使NLRP3、ASC及caspase-1蛋白表达下降。结论棕榈酸刺激肝细胞后,可以导致脂滴在肝细胞质中大量沉积,亦可以通过线粒体和NOX4途径导致肝细胞氧化应激反应,并因此促进细胞炎症小体的激活和IL-1β的分泌。Objective To evaluate the effect of palmitic acid （PA） on oxidative stress and activation of inflammasomes in hepatocytes. Methods To test the dose-dependent effect of PA on normal murine hepatocytes AML12, the cells were treated with 0, 0.15, 0.25 and 0.4 mmol/L of palmitic acid （PA）. The cells were also divided into blank control group, 0.25 mmol/L PA group and 0.25 mmol/L PA＋N-acetylcysteine （NAC） group to examine the effect of reactive oxygen species （ROS） on the activation of inflammasomes. After 24 h of treatment, lipid accumulation, total ROS, mitochondrial ROS, expression and localization of NOX4, and expressions of inflammasomes and IL-1βwere detected in the hepatocytes. Results Compared with the control cells, PA treatment of the cells significantly increased cytoplasmic lipid accumulation, concentrations of total ROS （12 463.09 ± 2.72 vs 6691.23 ± 2.45, P=0.00） and mitochondrial ROS （64.98 ± 0.94 vs 45.04 ± 0.92, P=0.00）, and the expressions of NOX4, NLRP3, ASC, caspase-1, and IL-1β（1603.52 ± 1.32 vs 2629.33 ± 2.57, P=0.00）. The mitochondria and NOX4 were found to be co-localized in the cytoplasm. NAC obviously reduced cellular ROS level stimulated by PA （7782.15±2.87 vs 5445.6±1.17, P=0.00） and suppressed the expressions of NLRP3, ASC and caspase- 1. Conclusion PA treatment can stimulate lipid accumulation in hepatocytes and induce oxidative stress through NOX4 and mitochondria pathway to activate inflammasomes and stimulate the secretion of IL-1β.

关 键 词：棕榈酸 NADPH氧化酶4 线粒体 氧化应激 炎症小体 

分 类 号：R965[医药卫生—药理学]