论文部分内容阅读
目的探讨bcl-2基因转染对培养的胰岛细胞凋亡和生物活性的影响。方法用腺病毒介导的基因转染方法把bcl-2转染入胰岛细胞,用RT-PCR和免疫细胞化学方法检测转染细胞中bcl-2的表达,原位末端标记(TUNEL)检测细胞凋亡,台盼蓝测定细胞活度,放免法测定培养液中胰岛素水平了解胰岛功能。结果转染的胰岛细胞中70%的细胞有bcl-2蛋白的表达,转染细胞的凋亡率为6%,对照组为22%,转染胰岛细胞的活度为91%,对照组为68%,转染胰岛细胞在高糖培养基中胰岛素水平高于对照组结论腺病毒介导的基因转染方法能成功转染胰岛细胞,bcl-2基因转染能降低细胞凋亡率,改善胰岛细胞功能。
Objective To investigate the effect of bcl-2 gene transfection on the apoptosis and bioactivity of cultured islet cells. Methods Transfection of bcl-2 into islet cells was performed by adenovirus-mediated gene transfection. The expression of bcl-2 in transfected cells was detected by RT-PCR and immunocytochemistry. TUNEL assay was used to detect the expression of bcl- Apoptosis and trypan blue were used to determine the cell viability. The level of insulin in culture medium was determined by radioimmunoassay to understand the function of islets. Results The expression of bcl-2 protein in 70% of transfected islet cells was 6% in the transfected cells, 22% in the control group and 91% in the transfected islet cells. The control group was 68%. The insulin level of transfected islet cells in high glucose medium was higher than that in control group. Conclusion The adenovirus-mediated gene transfection method can successfully transfect pancreatic islet cells. The bcl-2 gene transfection can reduce the apoptosis rate and improve Islet cell function.