摘要
目的探究对氧磷酶-1(Paraoxonase 1,PON1)与非酒精性脂肪肝(Nonalcoholic fatty liver disease,NAFLD)脂质代谢与胰岛素抵抗关系及机制。方法选取129例NAFLD患者和130非NAFLD患者为研究对象,收集患者年龄、身高、体重、腰围,血压等信息,检测血脂、空腹血糖(Fast Glucose)、胰高血糖素(Glucagon)、胰岛素(Fast Insulin)、评估胰岛素抵抗的稳态指数(homeostatic index of insulin resistance,HOMA-IR)、肝肾功能、脂肪因子等,LO2细胞接种12孔板培养24 h后分别加入0~1.2 mmol OA溶液处理24 h或10 ng/mL TNF-α处理6 h,以PEI转染空载质粒和PON1质粒1μg 24 h。使用尼罗红染料测定细胞内脂质含量。ELISA法检测PON1活性,蛋白质印迹检测PON1、p-IRS1、T-IRS1、p-AKT T-AKT、FAS、HMGCR、SREBP2和SREBP1C蛋白表达。结果NAFLD患者腰围、BMI、外周血ALT、AST、FFA、TG、TC、LDL-C、VLDL-C、空腹血糖、胰岛素、HOMA-IR、抵抗素、瘦素、IL-6和TNF-α显著高于非NAFLD,PON1活性、含量、HDL-C、胰高血糖素和脂联素显著降低;血PON1活性和含量与FFAs、TG、TC、LDL-C、VLDL-C、空腹血糖、胰岛素、HOMA-IR、抵抗素、瘦素、IL-6和TNF-α显著负相关,与HDL-C、胰高血糖素和脂联素显著正相关;OA组LO2细胞PON1的活性逐渐降低,且0.2~1.2 mM OA组LO2细胞PON1的活性显著低于Control组,过表达PON1不影响PON1活性,0.2~1.2 mM OA组LO2细胞内脂质含量显著高于Control组,显著低于OA组;TNF-α组LO2细胞PON1、p-IRS1、p-AKT、FAS、HMGCR、SREBP2和SREBP1C蛋白表达量相比于Control组显著降低,转染PON1质粒显著增加PON1,同时p-IRS1、p-AKT、FAS、HMGCR、SREBP2和SREBP1C蛋白表达量显著增加。结论PON1活性与含量和NAFLD血脂代谢及胰岛素抵抗显著相关,且可能通过调控p-IRS1/AKT通路影响胰岛素抵抗,并调控脂质代谢。
Objective To investigate the role and mechanism of paraoxonase 1(PON1)and lipid metabolism,insulin resistance in nonalcoholic fatty liver disease(NAFLD).Methods One hundred and twenty-nine NAFLD patients and 130 non-NAFLD individuals were enrolled in the study.The age,height,weight,waist circumference,blood pressure and other information were collected.Plasma blood lipids,fast glucose,and glucagon were detected by routine blood tests,homeostatic index of insulin resistance(HOMA-IR)was calculated,liver and kidney function were evaluated in biochemical assay,LO2 cells were inoculated in 12-well plates for 24 h and then added 0~1.2 mM OA solution was treated for 24 h or 10 ng/mL TNF-αfor 6 h,and the 1μg control and PON1 plasmid were transfected with PEI for 24 h.The intracellular lipid content was determined using a Nile Red dye.PON1 activity was detected by ELISA,and PON1,p-IRS1,T-IRS1,p-AKT T-AKT,FAS,HMGCR,SREBP2 and SREBP1C protein were detected by Western Blot.Results Waist circumference,BMI,peripheral blood ALT,AST,FFAs,TG,TC,LDL-C,VLDL-C,fasting blood glucose,insulin,HOMA-IR,resistin,leptin,IL-6 and TNF-αwere significantly higher in patients with NAFLD.In non-NAFLD,PON1 activity,content,HDL-C,glucagon and adiponectin were significantly reduced;PON1 activity and content were significantly negatively correlated with FFAs,TG,TC,LDL-C,VLDL-C,fasting blood glucose,insulin,HOMA-IR,resistin,leptin,IL-6 and TNF-α,and positively correlated with HDL-C,glucagon and adiponectin;PON1 activity in LO2 cells of OA group gradually decreased,and the activity of PON1 in LO2 cells of 0.2~1.2 mM OA group was significantly lower than that of control group.Overexpression of PON1 did not affect PON1 activity,the lipid content of LO2 cells in 0.2~1.2 mM OA group was significantly higher.In the control group,it was significantly lower than the OA group;the expression levels of PON1,p-IRS1,p-AKT,FAS,HMGCR,SREBP2 and SREBP1C in the TNF-αgroup were significantly lower than those in the control group,and the transfected PON1 plasmid w
作者
张荣
陈婧
黄江涛
吕海龙
杨一邨
王浩斌
张抒
ZHANG Rong;CHEN Jing;HUANG Jiang-tao;LU Hai-long;YANG Yi-cun;WANG Hao-bin;ZHANG Shu(Department of general surgery,Third People's Hospital of Chengdu,610031;Department of gastroenterology,Dujiangyan People′s Hospital,611830)
出处
《肝脏》
2020年第2期149-154,共6页
Chinese Hepatology
基金
国家自然基金(81702963)。
