黄芪多糖对高游离脂肪酸所致巨噬细胞脂质沉积及腺苷酸活化蛋白激酶-脂肪酸结合蛋白4通路的作用  

Influence of astragalus polysaccharides on macophage intracellular lipid accumulation induced by free fatty acid and adenosine monophosphate activated protein kinase-fatty acid binding protein 4 signaling

作　　者：胡阳黔[1] 李静[2] 刘坚[2] 王文峰[1] 宋杰[1] 

机构地区：[1]湖北医药学院附属东风医院消化内科,湖北省十堰市442000 [2]湖北医药学院基础医学院解剖教研室

出　　处：《中国医药》2014年第9期1388-1392,共5页China Medicine

基　　金：国家自然科学基金（30370673）;湖北省教育厅科学技术研究（B2013120）;湖北省十堰市科学技术研究与开发项目（ZD2012037）

摘　　要：目的：探讨黄芪多糖对高游离脂肪酸导致巨噬细胞内脂质沉积的作用及机制。方法将培养的单核源性巨噬细胞（ THP-1）分为对照组（加入与游离脂肪酸组等量的无水乙醇及牛血清清蛋白）、黄芪多糖组（培养液中加入黄芪多糖200 mg/L）、游离脂肪酸组（培养液中加入长链游离脂肪酸0．25 mmol/L）、游离脂肪酸加黄芪多糖组（培养液中加入黄芪多糖200 mg/L及长链游离脂肪酸0．25 mmol/L）及游离脂肪酸加黄芪多糖及腺苷酸活化蛋白激酶（AMPK）阻断剂（compound C）组（培养液中加入黄芪多糖200 mg/L、长链游离脂肪酸0．25 mmol/L及compound C 50μmol/L）。采用游离脂肪酸检测试剂盒和三酰甘油试剂盒分别检测游离脂肪酸及三酰甘油含量，油红O染色观察细胞内脂质蓄积，蛋白质印迹法检测细胞内AMPK、磷酸化腺苷酸活化蛋白激酶（p-AMPK）、脂肪酸结合蛋白4（FABP4）的蛋白表达情况。结果①游离脂肪酸组三酰甘油及游离脂肪酸含量[（3．35±0．79）mmol/L、（29．3±1．7）μmol/L]明显高于对照组[（0．81±0．19）mmol/L、（15．7±1．3）μmol/L]和游离脂肪酸加黄芪多糖组[（1．46±0．25）mmol/L、（15．5±2．1）μmol/L]，差异均有统计学意义（均P＜0．01）。②油红O染色结果：游离脂肪酸组脂质含量较对照组增多，游离脂肪酸加黄芪多糖组脂质含量较游离脂肪酸组减少，提示游离脂肪酸增加细胞内脂质蓄积，而黄芪多糖减少细胞内脂质蓄积。③蛋白质印迹法检测AMPK、p-AMPK、FABP4的蛋白表达情况发现，与对照组相比，黄芪多糖组各种蛋白表达无明显变化，游离脂肪酸组p-AMPK表达明显少于对照组和游离脂肪酸加黄芪多糖组[（54．3±3．0）％比（100．0±1．4）％、（96．5±3．9）％]，FABP4表达明显高于对照组和游离脂肪酸加黄芪多糖组[（138．3±2．3）％比（100．0±3．1）％、（72．Objective To investigate the effects of astragalus polysaccharide （ APS） on lipid accumula-tion in macrophages induced by free fatty acids （ FFA） and mechanism.Methods The cultured derived THP-1 monocyte macrophages were divided into 5 groups： the control group （ ethanol and bovine serum albumin , equal with FFA group）, APS group （APS 200 mg/L）, FFA group （long chain FFA 0.25 mmol/L）, FFA and APS group （APS 200 mg/L＋long chain FFA 0.25 mmol/L）, FFA ＋APS ＋amp activated protein kinase（AMPK） inhibitor （compound C） group （APS 200 mg/L＋long chain FFA 0.25 mmol/L＋compound C 50μmol/L）.The FFA detection kit and triacylglycerol （ TG） kit were used to detect FFA and TG .The intracellular lipid accumula-tion was analyzed by oil red O staining , and protein expressions of AMPK , phosphorylated AMPK （ p-AMPK） and fattyacidbindingprotein4（FABP4）wereanalyzedbywesternblotting.Results ①ThecontentsofTGandFFA were significantly higher in FFA group than in the control group [（3.35 ±0.79） mmol/L vs （0.81 ±0.19） mmol/L, （29.3 ±1.7） μmol/L vs （15.7 ±1.3） μmol/L]; the contents of TG and FFA were significantly decreased in FFA and APS group than those in FFA group [（1.46 ±0.25） mmol/L vs （3.35 ±0.79） mmol/L, （15.5 ± 2.1） μmol/L vs （29.3 ±1.7） μmol/L], the differences were statistically significant （all P〈0.01）.②The oil red O staining results indicated that lipid content was significantly increased in FFA group than in the control group;lipid content was significantly lower in FFA and APS group than in FFA group , which suggested that APS could reduce lipid accumulation induced by FFA .③The expression of AMPK , p-AMPK and FABP4 protein were no difference between APS group and control group .Compared with the control group , the FFA group decreased p-AMPK expression, and increased FABP4 expression [（54.3 ±3.0）%vs （100.0 ±1.4）%,（138.3 ±2.3）%vs （100.0 ±3.1）%];compared with the FFA group , the FFA and APS gr

关 键 词：黄芪多糖 游离脂肪酸 腺苷酸活化蛋白激酶 脂肪酸结合蛋白4 巨噬细胞 脂质沉积 

分 类 号：R589.2[医药卫生—内分泌]