钙非依赖性磷脂酶A2抑制剂减少肝细胞脂性凋亡及改善胰岛素抵抗的研究  

Effect of calcium-independent phospholipase A2 inhibitor in reducing hepatocyte lipoapoptosis and improving insulin resistance

作　　者：施红波 傅君芬[2] 黄邀[2] 刘利蕊[2] 

机构地区：[1]宁波市医疗中心李惠利东部医院,315040 [2]浙江大学医学院附属儿童医院内分泌科,杭州310051

出　　处：《中华肝脏病杂志》2017年第1期27-31,共5页Chinese Journal of Hepatology

基　　金：浙江省医学重点学科（创新学科）（11-CX24）;国家自然科学基金（81270938）

摘　　要：目的研究钙非依赖性磷脂酶A2（iPLA：）抑制剂在减少肝细胞脂性凋亡和改善胰岛素抵抗中的意义。方法28只雄性SD大鼠随机分成3组：I组（正常对照组）12只，普通饲料喂养18周；Ⅱ组（非酒精性脂肪性肝病模型组）8只，高脂饮食喂养18周；Ⅲ组（iPLA：抑制剂组）8只，高脂饮食喂养18周，第15周开始腹腔注射溴烯醇内酯（iPLA：抑制剂），每次150μg／kg，隔日1次，共14次。然后同时处死，称体质量、肝质量。检测各组血脂、血清酶学、空腹血糖、空腹胰岛素、游离脂肪酸、血iPLA，浓度，检测肝组织病理学变化，用末端脱氧核苷酸转移酶介导的脱氧三磷酸尿昔缺口末端标记法检测肝细胞凋亡水平并计算凋亡指数；定量PCR法检测肝iPL-RNA的表达。采用Student—Newman-Keuls检验方法及卡方检验的方法比较Ⅰ组和Ⅱ组、Ⅱ组和Ⅲ组间的各指标的差异，P〈0．05为差异有统计学意义。结果I组与Ⅱ组比较：甘油三酯（0．75±0．05）mmol／L比（1．20±0．13）mmol／L、胆固醇（1．50±0．12）mmol／L比（2．94±0．34）mmol/L、低密度脂蛋白（0．65±0．06）mmol／L比（1．30±0．16）mmol／L、游离脂肪酸（0．58±0．09）mEq／L比（0．80±0．20）mEq／L、空腹血糖（4．85±0．22）mmol／L比（6．94±0．65）mmol／L、空腹胰岛素（0．89±0．52）mmol／L比（1．29±0．52）mmol／L均明显上升，胰岛素敏感指数0．52±0．21比0．27±0．11，明显下降护〈0．05）；肝脏病理学检测结果提示Ⅱ组明显的炎症及脂肪变性，可见凋亡细胞，凋亡指数（0．58％±0．17％比39．69％±4．96％）明显增高垆〈0．05）；血沪LA，浓度（2．92±0．08）ng／ml比（3．28±0．14）ng／ml及肝iPLA2mRNA表达（1．07±0．18比7．68±0．49）明显增高护〈0．05）。Ⅱ组与Ⅲ组比较：肝细胞的凋亡减少，凋亡指数（39．69％±4．96％比24．80％±2�Objective To investigate the effect of calcium-independent phospholipase A2 （iPLA2） inhibitor in reducing hepatocyte lipoapoptosis and improving insulin resistance. Methods A total of 28 male Sprague-Dawley rats were randomly divided into the following three groups： 12 rats in group Ⅰ （normal control group） were given normal diet for 18 weeks; 8 rats in group Ⅱ （non-alcoholic fatty liver disease model group） were given high-fat diet for 18 weeks; 8 rats in group Ⅲ （iPLA2 inhibitor group） were given high-fat diet for 18 weeks and inlraperitoneal injection of the iPLA2 inhibitor bromoenol lactone 150 μg/kg once every other day since week 15 （14 times of injection in total）. All the rats were sacrificed at the same time, and body weight and liver weight were measured. Blood lipids, serum enzymes, fasting blood glucose, fasting insulin, free fatty acid, and serum iPLA2 concentration were measured in each group, and liver pathological changes were evaluated. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to measure the level of hepatocyte apoptosis and the apoptotic index was calculated. Quantitative PCR was used to measure the mRNA expression of iPLA2. The Student-Newman-Keuls test and the chi-square test were used for comparison of parameters between groups Ⅰ, Ⅱ, and Ⅲ P 〈 0.05 was considered statistically significant. Results Compared with group Ⅰ, groupⅡ had significant increases in triglyceride （0.75±0.05 mmol/L vs 1.20±0.13 mmol/L,P 〈 0.05）, cholesterol （1.50±0.12 mmol/L vs 2.94±0.34 mmol/L, P 〈 0.05）, low-density lipoprotein （0.65±0.06 mmol/L vs 1.30±0.16 mmol/L, P 〈 0.05）, free fatty acid （0.58±0.09 mEq/L vs 0.80±0.20 mEq/L, P 〈 0.05）, fasting blood glucose （4.85±0.22 mmol/L vs 6.94±0.65 mmol/L, P 〈 0.05）, and fasting insulin （0.89±0.52 mmol/L vs 1.29±0.52 mmol/L, P 〈 0.05）, and a significant reduction in the insulin sensitivity index （0.52±0.21 vs 0.27±0.11, P 〈 0.05）; group Ⅱ

关 键 词：脂肪肝 非酒精性 大鼠 胰岛素 

分 类 号：R575[医药卫生—消化系统]