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褪黑素受体激动剂Neu-P11对3T3-L1小鼠脂肪细胞蛋白激酶B及其磷酸化的影响
李秀平1,蔡世昌2*,尹卫东3,张素君4,胡蓉1,李兴2,Moshe,Laudon5
0
(1. 湖南医药学院医学检验系, 怀化 418000;
2. 湖南医药学院基础医学部, 怀化 418000;
3. 南华大学医学院心血管疾病研究所, 衡阳 412000;
4. 南华大学实验动物学部, 衡阳 412000;
5. Neurim Pharmaceuticals Ltd., Tel-Aviv 69710, Israel
*通信作者)
摘要:
目的:研究褪黑素受体激动剂Neu-P11对胰岛素抵抗3T3-L1脂肪细胞PKB、P-PKB的蛋白表达变化,并研究其变化机制。方法:采用“鸡尾酒”法培养与分化3T3-L1前脂肪细胞,建立胰岛素抵抗细胞模型,用Western-blot检测药物作用前后PKB, P-PKB的蛋白质表达变化。结果:胰岛素抵抗组的脂肪细胞PKB, P-PKB的表达明显低于对照组(P < 0.05)。药物Mel, Neu-P11均能明显增加PKB、P-PKB蛋白表达量(P < 0.05)。Luzindole能够阻断Mel, Neu-P11增加PKB的作用(P < 0.05),也能够阻断Mel增加P-PKB的作用(P < 0.05),但不能阻断Neu-P11增加P-PKB的作用。结论:褪黑素、Neu-P11可以提高胰岛素抵抗组细胞PKB, P-PKB的蛋白表达。褪黑素和Neu-P11可能是通过与褪黑素受体2结合发挥增加PKB蛋白表达作用的;褪黑素可能是通过与褪黑素受体2结合来增加P-PKB蛋白表达的。而Neu-P11提高P-PKB的蛋白表达可能是通过其他的途径,还需要进一步研究。
关键词:  Neu-P11  褪黑素  胰岛素抵抗  蛋白激酶B  Luzindole
DOI:10.3724/SP.J.1008.2015.00802
投稿时间:2015-03-07修订日期:2015-04-21
基金项目:湖南省教育厅科学研究项目(14C0909),湖南医药学院科学研究项目(2014KY02).
Effect of melatonin receptor agonist Neu-P11 on phosphorylation of protein kinase B in mouse 3T3-L1 adipocytes
LI Xiu-ping1,CAI Shi-chang2*,YIN Wei-dong3,ZHANG Su-jun4,HU Rong1,LI Xing2,Moshe Laudon5
(1. Department of Laboratory, Hunan University of Medicine, Huaihua 418000, Hunan, China;
2. Division of Basic Medical Sciences, Hunan University of Medicine, Huaihua 418000, Hunan, China;
3. Institute of Cardiovascular Disease, University of South China Medical School, Hengyang 412000, Hunan, China;
4. Division of Experimental Animal, University of South China, Hengyang 412000, Hunan, China;
5. Neurim Pharmaceuticals Ltd., Tel-Aviv 69710, Israel
*Corresponding author)
Abstract:
Objective: The objective of present study was to examine the changes of melatonin receptor agonist Neu-P11 on expression of PKB and P-PKB in 3T3-L1 adipocytes and to research the variation mechanisms.Methods: 3T3-L1 pre-adipocytes were cultivated and differentiated in adipogenic cocktail by IBMX, DEX and insulin; Confirm the establishment of insulin resistance model. The changes of PKB and P-PKB protein expressions before and after drug action were detected by Western blot; Results: Expression of PKB and P-PKB reduced significantly by comparison with IS (P < 0.05). Melatonin and Neu-P11 can increase obviously the expression of PKB and P-PKB (P < 0.05). But Luzindole can block the effect of melatoninl, Neu-P11 increasing PKB (P < 0.05), will also be able to block that the melatonin increase P- PKB (P < 0.05), but it can"t block that Neu-P11 increases P - PKB .
Key words:  Neu-P11  melatonin  insulin resistance  protein kinase B  Luzindole