腺病毒36感染对人脂肪源性间充质干细胞PPARγ和adiponectin基因表达的影响

发布时间：2019-01-27 10:56

【摘要】：目的探讨腺病毒36感染对人脂肪源性间充质干细胞过氧化物酶体增殖物激活受体(peroxisome proliferator-activated receptor,PPARγ)和脂联素(adiponectin)基因表达的影响。方法无菌条件下获取人皮下脂肪组织、I型胶原酶消化并收集人脂肪源性间充质干细胞(human adipose-derived mesenchymal stem cell,h AMSC),细胞传至第3代(P3代)对h AMSC进行定向成脂诱导,并进行油红O染色;流式细胞仪检测h AMSC表面免疫表型;人类腺病毒36(adenouirus-36,Ad36)感染h AMSC,采用荧光实时定量PCR(Real time-PCR)方法检测Ad36感染的h AMSC PPARγ和adiponectin基因表达的变化规律。结果 (1)h AMSC在体外呈梭形生长,且能稳定传代;(2)细胞在加入成脂诱导剂后可定向分化成脂肪细胞;(3)流式细胞术检测结果显示h AMSC免疫表型:CD105(93.0±1.3)%,CD44(99.6±0.3)%,CD34(0.2±0.1)%,CD45(0.2±0.3)%,CD31(0.3±0.3)%,CD29(3.4±0.2)%;(4)Ad36感染h AMSC后细胞内脂滴逐渐增多、增大。PPARγ和adiponectin基因在Ad36感染h AMSC后的第1天开始表达,感染后第2~6天较0 MOI组显著升高(P0.05),至感染后第8天10 MOI剂量组表达水平开始下降。结论 Ad36使h AMSC定向分化成脂肪细胞,并上调了PPARγ和adiponectin基因的表达。
[Abstract]:Objective to investigate the effects of adenovirus 36 infection on peroxisome proliferator activated receptor (peroxisome proliferator-activated receptor,PPAR 纬) and adiponectin (adiponectin) gene expression in human adipose derived mesenchymal stem cells. Methods Human subcutaneous adipose tissue was obtained under aseptic condition. Human adipose-derived mesenchymal stem cells (human adipose-derived mesenchymal stem cell,h AMSC), cells) were digested by type I collagenase and then transferred to the 3rd generation (P3 generation) to induce direct adipogenesis of AMSC. And oil red O staining; The surface immunophenotype of h AMSC was detected by flow cytometry, and the expression of h AMSC PPAR 纬 and adiponectin genes in h AMSC, infected by human adenovirus 36 (adenouirus-36,Ad36) was detected by fluorescence real time quantitative PCR (Real time-PCR. Results (1) h AMSC grew fusiform in vitro and could be subcultured stably. (2) adipocytes could be differentiated into adipocytes after adding lipogenic inducer. (3) the results of flow cytometry showed that CD105 was (93.0 卤1.3)%, CD44 was (99.6 卤0.3)%, CD34 was (0.2 卤0.1)%, CD45 was (0.2 卤0.3)%, CD31 was (0.3 卤0.3)%. CD29 (3.4 卤0.2)%; (4) the lipid droplets in the cells increased gradually after Ad36 infection with AMSC, and the expression of PPAR 纬 and adiponectin genes began on the first day after Ad36 infection with AMSC, and increased significantly on the 2nd day after Ad36 infection compared with 0 MOI group (P0.05). The expression level of 10 MOI dose group began to decrease on the 8th day after infection. Conclusion Ad36 can induce h AMSC to differentiate into adipocytes and up-regulate the expression of PPAR 纬 and adiponectin genes.
【作者单位】：新疆医科大学基础医学院生物化学教研室;新疆维吾尔自治区人民医院;
【基金】：国家自然科学基金项目(30960363) 博士(后)科研启动基金(2012-7)
【分类号】：R373

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