慢病毒载体介导的脂肪细胞SOCS-3RNA干扰及对瘦素反应的研究  

Research on the Effect of SOCS-3 RNA Interference Mediated by Lentiviral Vector in Adipocytes with Leptin Treatment

作　　者：刘莉[1] 刘亚莉[2] 马爽[3] 赵越[1] 安丽[1] 于飞[1] 任亚浩[1] 杨军[1] 

机构地区：[1]中国医科大学公共卫生学院,中国辽宁沈阳110001 [2]辽宁中医药大学职业技术学院,中国辽宁沈阳110101 [3]中国医科大学附属第一医院,中国辽宁沈阳110001

出　　处：《生命科学研究》2010年第5期377-380,共4页Life Science Research

基　　金：国家自然科学基金资助项目(30500409)

摘　　要：体内存在瘦素抵抗是大多数肥胖者的共同特征,但如何缓解瘦素抵抗目前还无有效方法.通过应用慢病毒载体介导的RNA干扰技术对大鼠脂肪细胞中瘦素信号转导通路的负反馈抑制因子—细胞因子信号转导抑制因子-3(suppressor of cytokine signaling 3,SOCS-3)基因进行敲减,观察其对大鼠重组瘦素作用的反应.研究发现,阴性对照慢病毒和SOCS-3shRNA慢病毒对成熟脂肪细胞的感染率约为80%.利用real-time PCR和Westernblot法检测发现SOCS-3 shRNA慢病毒通过在成熟脂肪细胞内表达shRNA而造成SOCS-3基因在mRNA和蛋白水平的敲减,有效率分别为72%和57%;经50nmol/L瘦素处理6h后,感染阴性对照病毒的脂肪细胞SOCS-3mRNA表达有明显升高(P<0.05),而感染SOCS-3 shRNA慢病毒的脂肪细胞SOCS-3mRNA表达则无明显变化(P>0.05).结果表明,利用SOCS-3shRNA慢病毒去敲减脂肪细胞中SOCS-3的表达可能对于解除肥胖者外周瘦素抵抗具有一定的作用.Leptin resistance is very common in obese people.However there has no effective methods in relieving leptin resistance.A negative feedback inhibition factor （suppressor of cytoki-ne signaling 3,SOCS-3） in leptin signaling transduction pathway was knocked-down by using lentiviral vector-mediated RNA interference in rat adipocytes.The effect of adipocytes with or without SOCS-3 knock-down treated with rat recombinant leptin was observed.The infection efficiency of both negative lentivirus and SOCS-3 shRNA lentivirus was 80%.SOCS-3 shRNA lentivirus could inhibit SOCS-3 mRNA and protein significantly through expressing shRNA in mature adipocytes by using real-time PCR and western blot determination,and the inhibition efficiency was 72% and 57%,respectively.After 6 h treatment with 50 nmol /L leptin,the expression of SOCS-3 mRNA in negative lentivirus group increased significantly （P 0.05）,while the expression of SOCS-3 mRNA in SOCS-3 shRNA lentivirus group was no change（P0.05）.The results indicated that the inhibition of SOCS-3 in adipocytes by using SOCS-3 shRNA lentivirus perhaps contributed to relieve peripheral leptin resistance of obese people.

关 键 词：细胞因子信号转导抑制因子-3 RNA干扰 慢病毒载体 瘦素 脂肪细胞 

分 类 号：Q291[生物学—细胞生物学]