中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2014年
9期
1658-1662
,共5页
细胞凋亡%SOCS1%胰岛细胞
細胞凋亡%SOCS1%胰島細胞
세포조망%SOCS1%이도세포
Apoptosis%SOCS1%Islet cell
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。
目的：觀察增加胰島細胞SOCS1錶達後胰島細胞功能及抗凋亡能力的變化。方法通過膽總管插管、逆行灌註膠原酶P溶液的方法分離大鼠胰島，採用Ficoll密度梯度離心法純化胰島，採用AdMax法構建腺病毒載體Ad5 F35-SOCS1併轉染大鼠胰島細胞，使用Western blot法檢測轉染胰島細胞的SOCS1錶達水平，通過胰島的糖刺激指數檢測大鼠胰島細胞的胰島素釋放功能，通過加入凋亡誘導劑及應用流式細胞儀檢測凋亡細胞的比例。結果與對照組及未轉染胰島細胞相比，經Ad5 F35-SOCS1轉染的大鼠胰島細胞在具有高錶達SOCS1的特性時其正常的胰島素釋放功能不受影響（三組的糖刺激指數分彆為3.98±0.45、4.06±0.61和3.53±0.39，P＞0.05），經凋亡誘導劑誘導後其凋亡細胞數明顯低于對照組及未轉染組，三組細胞凋亡率分彆為（8.89±4.03）%、（24.60±6.88）%和（21.14±5.12）%，P＜0.05。結論增加胰島細胞 SOCS1錶達不影響胰島細胞的功能，併能有效減少細胞的凋亡。
목적：관찰증가이도세포SOCS1표체후이도세포공능급항조망능력적변화。방법통과담총관삽관、역행관주효원매P용액적방법분리대서이도，채용Ficoll밀도제도리심법순화이도，채용AdMax법구건선병독재체Ad5 F35-SOCS1병전염대서이도세포，사용Western blot법검측전염이도세포적SOCS1표체수평，통과이도적당자격지수검측대서이도세포적이도소석방공능，통과가입조망유도제급응용류식세포의검측조망세포적비례。결과여대조조급미전염이도세포상비，경Ad5 F35-SOCS1전염적대서이도세포재구유고표체SOCS1적특성시기정상적이도소석방공능불수영향（삼조적당자격지수분별위3.98±0.45、4.06±0.61화3.53±0.39，P＞0.05），경조망유도제유도후기조망세포수명현저우대조조급미전염조，삼조세포조망솔분별위（8.89±4.03）%、（24.60±6.88）%화（21.14±5.12）%，P＜0.05。결론증가이도세포 SOCS1표체불영향이도세포적공능，병능유효감소세포적조망。
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.
