Berberine reverses free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKβ
Berberine reverses free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKβ
复制标题
DOI：
10.3748/wjg.14.876
复制
发表时间：
2008-02-14
影响因子：
4.3
通讯作者：
Wang, Kai-Fu
中科院分区：
文献类型：
--
作者：
Yi, Ping;Lu, Fu-Er;Wang, Kai-Fu
目的：探讨小檗碱改善游离脂肪酸诱导的3T3-L1脂肪细胞胰岛素抵抗的作用及其分子机制。方法：在培养液中加入0.5 mmol/L棕榈酸，建立3T3-L1脂肪细胞胰岛素抵抗模型。同时进行小檗碱治疗。葡萄糖摄取率采用2-脱氧-[H-3]- d -葡萄糖法测定。Western blotting检测IkB激酶β (IKK β) Ser（181）磷酸化水平、胰岛素受体底物-1(IRS-1) Ser（307）磷酸化水平、IKK β、IRS-1、核转录因子kappaB p65 （NF-kappa B p65）、磷脂酰肌醇-3激酶p85 （PI-3K p85）和葡萄糖转运蛋白4 （GLUT4）蛋白表达水平。通过激光共聚焦扫描显微镜（CLSM）观察NF-kappa B p65蛋白在脂肪细胞内的分布。结果：棕榈酸干预24 h后，胰岛素刺激的3T3-L1脂肪细胞葡萄糖转运被抑制67%。同时，IRS-1和PI-3K p85蛋白表达降低，IKK β Ser（181）和IRS-1 Ser（307）磷酸化水平升高，NF-kappa B p65蛋白核易位升高。而上述指标均被小檗碱逆转，表明存在胰岛素抵抗，但本研究并未改变GLUT4、IKK β和总NF-kappa B p65蛋白的表达。结论：小檗碱可改善FFAs诱导的3T3-L1脂肪细胞胰岛素抵抗。小檗碱通过靶向IKK β逆转游离脂肪酸诱导的3T3-L1脂肪细胞胰岛素抵抗。(c) 2008年版权所有。
AIM: To investigate the effects and molecular mechanisms of berberine on improving insulin resistance induced by free fatty acids (FFAs) in 3T3-L1 adipocytes.METHODS: The model of insulin resistance in 3T3-L1 adipocytes was established by adding palmic acid (0.5 mmol/L) to the culture medium. Berberine treatment was performed at the same time. Glucose uptake rate was determined by the 2-deoxy-[H-3]-D-glucose method. The levels of IkB kinase beta (IKK beta) Ser(181) phosphorylation, insulin receptor substrate-1(IRS-1) Ser(307) phosphorylation, expression of IKK beta, IRS-1, nuclear transcription factor kappaB p65 (NF-kappa B p65), phosphatidylinositol-3-kinase p85 (PI-3K p85) and glucose transporter 4 (GLUT4) proteins were detected by Western blotting. The distribution of NF-kappa B p65 proteins inside the adipocytes was observed through confocal laser scanning microscopy (CLSM).RESULTS: After the intervention of palmic acid for 24 h, the insulin-stimulated glucose transport in 3T3-L1 adipocytes was inhibited by 67%. Meanwhile, the expression of IRS-1 and PI-3K p85 protein was reduced, while the levels of IKK beta Ser(181) and IRS-1 Ser(307) phosphorylation, and nuclear translocation of NF-kappa B p65 protein were increased. However, the above indexes, which indicated the existence of insulin resistance, were reversed by berberine although the expression of GLUT4, IKK beta and total NF-kappa B p65 protein were not changed during this study.CONCLUSION: Insulin resistance induced by FFAs in 3T3-L1 adipocytes can be improved by berberine. Berberine reversed free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKK beta. (C) 2008 WJG. All rights reserved.