药根碱多重调控脂肪胰岛素抵抗细胞葡萄糖转运蛋白的降糖效应研究
投稿时间:2017-10-23  修订日期:2017-11-02  责任编辑:  点此下载全文
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作者中文名作者英文名单位中文名单位英文名E-Mail
朱水兰 Zhu Shuilan 江西中医药大学 Jiangxi University of Traditional Chinese Medicine 604985245@qq.com 
雷婷 Lei ting 江西省中医药大学 Jiangxi Province Key Laboratory of TCM Etiopathogenisis,Research Center for Differentiation and Development of TCM Basic Theory, Jiangxi University of Traditional Chinese Medicine  
高旭 Gao Xu 江西中医药大学 Jiangxi Province Key Laboratory of TCM Etiopathogenisis,Research Center for Differentiation and Development of TCM Basic Theory, Jiangxi University of Traditional Chinese Medicine  
涂珺* Tu Jun 江西中医药大学 Jiangxi University of Traditional Chinese Medicine jtu2012@163.com 
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
中文摘要:研究药根碱对脂肪细胞胰岛素抵抗(IR)糖代谢的调节及降糖效应机制。本实验采用3T3-L1前脂细胞诱导分化的成熟脂肪细胞,1 μmol·L-1地塞米松诱导建立稳定IR模型。细胞给药分组如下:正常组、IR模型组、罗格列酮阳性组、药根碱组(0.5,1,5,10,20 μmol·L-1)。以葡萄糖氧化酶法检测不同时间点(12,24,30,36,48 h)3T3-L1细胞培养液葡萄糖含量,甘油磷酸氧化酶法测定细胞内TG含量,CCK-8法检测细胞活力;Western blot 检测胰岛素受体底物2(IRS2),磷脂酰肌醇3-激酶调节亚基1(PI3KR1),磷酸化蛋白激酶B(p-AKT(Ser473)),磷酸化腺苷酸活化蛋白激酶(p-AMPK(Thr172)),葡萄糖转运蛋白(GLUT4/1/2),等蛋白的表达水平。研究结果表明,与正常组相比,IR模型组葡萄糖消耗量显著降低(P<0.01);与IR组比较,0.5,1,5,10,20 μmol·L-1药根碱给药36和48 h能使IR-3T3-L1脂肪细胞葡萄糖消耗量显著增加(P<0.01),确定药根碱最佳作用时间为48 h。1,5,10,20 μmol·L-1药根碱作用48 h后,3T3-L1脂肪细胞内TG含量降低(P<0.05)。各浓度药根碱不同程度显著上调IRS2,PI3KR1,p-AKT,p-AMPK,GLUT4/1/2等蛋白表达水平(P<0.01)。结果表明药根碱明显增加 IR-3T3-L1脂肪细胞葡萄糖摄取和消耗,降低胞内TG含量,激活胰岛素降糖通路IRS2/PI3KR1/p-AKT/GLUT4,增加p-AMPK表达激活GLUT4/1/2来减轻脂肪IR
中文关键词:药根碱  葡萄糖转运蛋白  降糖效应  胰岛素抵抗  3T3-L1脂肪细胞
 
Jatrorrhizine regulates the GLUTs with multiple manners for anti-hyperglycemic effect in the insulin-resistance 3T3-L1 adipocytes
Abstract:To investigate the anti-hyperglycemic effect and relative mechanism by regulating the glucose metabolism in insulin-resistance (IR)-3T3-L1 adipocytes. The 3T3-L1 preadipocytes was used to induce mature adipocytes, then was established as a stable IR model with 1 μmol·L-1 dexamethasone. The adipocytes were divided into normal group, IR model group, rosiglitazone positive group and jatrorrhizine group (0.5, 1, 5, 10, 20 μmol·L-1). After different time points (12, 24, 30, 36, 48 h) treatment, glucose consumptions of 3T3-L1 adipocytes was detected by the glucose oxidase peroxidase method and TG content were measured by glycerol phosphate oxidase method, whereas cell viability was detected by CCK-8 assay. Furtherly, the protein expression levels of insulin receptor substrate 2(IRS2), phosphinositide-3-kinase regulatory subunit 1(PI3KR1), phosphorylated protein kianse B (p-AKT(Ser473)), phosph-AMP-activated protein kianse(p-AMPK(Thr172)), glucose transporter type 4/1/2(GLUT4/1/2) were detected by western blot assay. The results showed that compared with the normal group, the glucose consumption of IR model group decreased (P<0.01); whereas 0.5, 1, 5, 10, 20 μmol·L-1 jatrorrhizine could elevate IR-3T3-L1 cells glucose consumption (P<0.01) for 36 h and 48 h administration when compared with IR group. The optimal administration time of anti-hyperglycemic effect was 48 h according to time-course study. 1, 5, 10, 20 μmol·L-1 of jatrorrhizine decreased the TG content in 3T3-L1 adipocytes for 48 h administration (P<0.05). The protein expression levels of IRS2, PI3KR1, p-AKT(Ser473), p-AMPK(Thr172), GLUT4/1/2 could be significantly up-regulate by different concentrations of jatrorrhizine (P<0.01). The results showed that jatrorrhizine could increase glucose uptake with elevated glucose consumption, whereas reduce intracellular TG content in IR-3T3-L1 adipocytes. Moreover, it could intervene classic insulin signal pathway IRS2/PI3KR1/p-AKT/GLUT4 and increase AMPK protein phosphorylation level for the activation of GLUT1/4 for insulin sensibility. Thus, jatrorrhizine could effectively regulate the GLUTs with multiple manners for anti-hyperglycemic effect.
keywords:jatrorrhizine  GLUTs  anti-hyperglycemic effect  insulin resistance (IR)  3T3-L1 adipocytes
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