棕榈酸诱导BRL-3A细胞产生胰岛素抵抗的作用机制  

Mechanism of Palmitic Acid Induces Insulin Resistance in BRL-3A Cells

作　　者：崔敏萱 沈歆 盖守昌 井娟 王力彬 张生勇[2] 刘雪英[2] CUI Minxuan;SHEN Xin;GAI Shouchang;JING Juan;WANG Libin;ZHANG Shengyong;LIU Xueying(Fourth Group of Students,School of Basic Medicine,the Air Force Medical University of PLA,Xi'an 710032,China;Teachings and Research Section of Pharmaceutical Chemistry,Department of Pharmacy,the Air Force Medical University of PLA,Xi'an 710032,China)

机构地区：[1]中国人民解放军空军军医大学基础医学院四大队,西安710032 [2]中国人民解放军空军军医大学药学系药物化学教研室,西安710032

出　　处：《医药导报》2021年第5期575-581,共7页Herald of Medicine

基　　金：陕西省重点研发计划(2017KW-047)。

摘　　要：目的建立BRL-3A细胞系胰岛素抵抗模型,筛选棕榈酸诱导的最佳条件,并探究其作用机制。方法采用CCK8法确定棕榈酸对BRL-3A细胞增殖的影响;采用葡萄糖检测试剂盒检测葡萄糖含量,确定不同浓度的棕榈酸对BRL-3A细胞摄取葡萄糖的影响;采用免疫印迹法(Western blotting)检测胰岛素受体底物1(IRS-1)、磷脂酰肌醇-3'-羟基激酶(PI3K)、磷酸化丝氨酸-苏氨酸激酶(p-AKT)、丝氨酸-苏氨酸激酶(AKT)、葡萄糖转运蛋白4(GLUT4)的蛋白表达。结果0.2 mmol·L^(-1)棕榈酸作用36 h对BRL-3A细胞增殖无影响;0.4 mmol·L^(-1)棕榈酸诱导6 h、0.25和0.2 mmol·L^(-1)棕榈酸诱导12 h,BRL-3A细胞开始出现明显胰岛素抵抗,移除刺激后48 h内仍然能够抑制胰岛素刺激状态下的糖消耗;在蛋白水平,棕榈酸降低了IRS-1、PI3K、p-AKT、GLUT4在BRL-3A细胞内的表达。结论高浓度棕榈酸能够通过抑制IRS-1等靶蛋白表达诱导BRL-3A细胞产生胰岛素抵抗。Objective To establish a cell model of insulin resistance in BRL-3A cells,and to elucidate the underlying mechanisms of palmitic acid(PA).Methods Using CCK8 method to observe the effect of different concentration and time of PA on proliferation in BRL-3A cells.The glucose consumption were detected at different concentration and time points by glucose detection kit.The expression of insulin receptor substance 1(IRS-1),phosphoinositide-3-kinase(PI3K),phosphorylated serine-threonine kinase(p-AKT),serine-threonine kinase(AKT),glucose transporter 4(GLUT4)in BRL-3A cells were detected by Western blotting.Results The 0.2 mmol·L^(-1) PA had no effect on proliferation in BRL-3A cells within 36 h.The insulin resistance model of BRL-3A cells can be successful established by incubating in cultured medium with PA in 3 conditions:0.4 mmol·L^(-1) PA for 6 h,0.25 mmol·L^(-1) PA for 12 h,0.2 mmol·L^(-1) PA for 12 h.And insulin stimulated glucose transport was inhibited by high concentration of PA in BRL-3A cells.Palmitic acid decreased IRS-1,PI3K,p-AKT,GLUT4 expression in BRL-3A cells.Conclusion High concentration of palmitic acid may induce insulin resistance in BRL-3A cells via inhibiting the expression of IRS-1 and other target proteins.

关 键 词：棕榈酸 胰岛素抵抗 胰岛素受体底物1 

分 类 号：R977[医药卫生—药品] R587.1[医药卫生—药学]