摘要
目的观察增加胰岛细胞SOCS1表达后胰岛细胞功能及抗凋亡能力的变化。方法通过胆总管插管、逆行灌注胶原酶P溶液的方法分离大鼠胰岛,采用Ficoll密度梯度离心法纯化胰岛,采用AdMax法构建腺病毒载体Ad5 F35-SOCS1并转染大鼠胰岛细胞,使用Western blot法检测转染胰岛细胞的SOCS1表达水平,通过胰岛的糖刺激指数检测大鼠胰岛细胞的胰岛素释放功能,通过加入凋亡诱导剂及应用流式细胞仪检测凋亡细胞的比例。结果与对照组及未转染胰岛细胞相比,经Ad5 F35-SOCS1转染的大鼠胰岛细胞在具有高表达SOCS1的特性时其正常的胰岛素释放功能不受影响(三组的糖刺激指数分别为3.98±0.45、4.06±0.61和3.53±0.39,P>0.05),经凋亡诱导剂诱导后其凋亡细胞数明显低于对照组及未转染组,三组细胞凋亡率分别为(8.89±4.03)%、(24.60±6.88)%和(21.14±5.12)%,P<0.05。结论增加胰岛细胞SOCS1表达不影响胰岛细胞的功能,并能有效减少细胞的凋亡。
Objective To investigate the change of pancreas islet function and apoptosis of islet cells after increasing the SOCS1 expression in islet. Methods Pancreatic islets were isolated by collagenase P digestion and purified by discontinuous Ficoll density gradient centrifugation. Then transfected the purified islet cells by adenovirus vector Ad5 F35-SOCS1, which was constructed by AdMax method. The expression of SOCS1 in the transfected islets were analyzed by Western blot, and the insulin release function was tested by glucose stimulation index. After incubated with apoptosis inducers （recombinant rat TNF-α and cycloheximide）, islet cells were checked for apoptosis by flow cytometry. Results Islet cells, which transfected by Ad5 F35-SOCS1, expressed more SOCS1 and resistant to apoptosis, while remain the normal insulin release function, compared to the control group and un-transfected group. The glucose stimulation index in each group showed no difference （3.98±0.45, 4.06±0.61 and 3.53±0.39, P〉0.05）. The apoptosis cells rate in Ad5 F35-SOCS1 transfected group, which was （8.89±4.03）%, was greatly decreased compared to Ad5 F35-EGFP transfected group （24.60±6.88）%and blank group （21.14±5.12）%（P〈0.05）. Conclusion Up-regulation of SOCS1 decreased the apoptosis level of islet cells without affected its insulin release function.
出处
《中华临床医师杂志（电子版）》
CAS
2014年第9期55-58,共4页
Chinese Journal of Clinicians(Electronic Edition)
