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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(P<0.05),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。
Objective To investigate the effect of adenovirus 36 infection on the gene expression of peroxisome proliferator-activated receptor (PPARγ) and adiponectin in human adipose-derived mesenchymal stem cells. Methods Human subcutaneous adipose tissue was obtained under sterile conditions. Human adipose-derived mesenchymal stem cells (h AMSCs) were digested with type I collagenase and collected. The cells were passaged to passage 3 (P3) h AMSCs were induced to adipocyte and oily red O staining. Flow cytometry was used to detect the immunophenotype of h AMSC surface. Human Adenovirus-36 (Ad36) cells were infected with h AMSC. Real-time PCR -PCR) was used to detect the changes of PPARγ and adiponectin gene expression in Ad36-infected h AMSCs. Results (1) h AMSC grew spindle-shaped in vitro and could stably passaged; (2) Cells were differentiated into adipocytes after adding adipogenic inducers; (3) Flow cytometry results showed that h AMSC immunization table (93.0 ± 1.3)%, CD44 (99.6 ± 0.3)%, CD34 (0.2 ± 0.1)%, CD45 (4) After Ad36 infected with hMSC, the number of lipid droplets in the cells gradually increased and increased. PPARγ and adiponectin genes began to express on the 1st day after Ad36 infected with hMSC, and were significantly increased from the 2nd to the 6th day after infection (P <0.05), and began to express on the 8th day after infection decline. Conclusion Ad36 directs h AMSCs into adipocytes and up-regulates the expression of PPARγ and adiponectin genes.