【作者】 王淼；

【导师】 邹大进；

【作者基本信息】 第二军医大学 ， 内分泌与代谢病， 2007， 博士

【摘要】 目的明确蛋白酪氨酸磷酸酶1B(PTP-1B)在人脂肪干细胞(hADSC)分化中的作用及机制。方法1、hADSC分离培养并鉴定。2、以不同浓度UA(PTP-1B特异性抑制剂)和原钒酸钠(SV,PTP广谱抑制剂)干预hADSC分化,检测油红染色、脂肪酸结合蛋白4(FABP4)mRNA。3、以UA孵育hADSC测定PTP-1B活性和胰岛素受体(IR)β亚基磷酸化水平;检测UA干预下诱导1-10天PTP-1B、PPARλ、C/EBPα和C/EBPβ表达。结果1、hADSC符合干细胞特性。2、UA组脂滴增多、增大,FABP4表达随UA剂量增大而升高,呈剂量依赖性;SV组未见脂滴。3、UA抑制PTP-1B活性,IRβ亚基磷酸化水平、PPARγ和C/EBPα表达增高,C/EBPβ表达高峰前移。结论1、原代培养hADSC成功。2、抑制PTP-1B增强了胰岛素信号转导,PPARγ和C/EBPα表达增高并提前激活C/EBPβ,促进hADSC分化。3、抑制PTP可抑制hADSC分化。更多还原

【Abstract】 Objective: to study effects of protein tyrosine phosphatase 1B (PTP-1B) on differentiation of human adipose-derived stem cell(hADSC). Methods: 1. hADSC were detected according to stem cell’s characteristics. 2.hADSC differentiation was intervented by UA(PTP-1B inhibitor) and sodium orthovanadate(SV, PTP inhibitor). Oil red O staining and FABP4 mRNA were detected. 3. PTP-1B and IR were extracted from hADSC that incubated with UA to detect PTP-1B activity and lever of phosphorylated IRβsubunit. Expression of PTP-1B, PPARγ, C/EBPαand C/EBPβmRNA were detected from 1st to 10th day of induction. Results: 1.hADSC accorded with stem cell’s characteristics. 2. Lipid droplets in UA groups were more and bigger and FABP4 increased according as UA dose. No lipid droplet in SV group. 3. UA inhibited PTP-1B activity, made phosphorylated IRβsubunit, PPARγand C/EBPαexpress higher, and C/EBPβearlier. Conclusions: 1. hADSC were cultured successfully. 2. Inhibiton of PTP-1B enhances insulin signal and expression of PPARγand C/EBPα, and makes C/EBPβexpress earlier, which promote differentiation of hADSC. 3.Inhibition of PTP inhibits differentiation of hADSC.更多还原