天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
11期
1069-1072
,共4页
黎金凤%林安华%邓颖%霍亚南%刘精东%吴明斌%王晨秀
黎金鳳%林安華%鄧穎%霍亞南%劉精東%吳明斌%王晨秀
려금봉%림안화%산영%곽아남%류정동%오명빈%왕신수
基质细胞衍生因子-1%内皮祖细胞%PI3K/AKT%糖尿病
基質細胞衍生因子-1%內皮祖細胞%PI3K/AKT%糖尿病
기질세포연생인자-1%내피조세포%PI3K/AKT%당뇨병
stromal cell-derived-factor-1%endothelial progenitor cells%phosphoinositide3-kinase/protein kinase B%diabetes
目的：观察基质细胞衍生因子-1（SDF-1）对糖尿病外周血内皮祖细胞（EPCs）功能的影响，探讨SDF-1对EPCs的影响是否与PI3K/AKT信号通路有关。方法采集糖尿病患者（DM组）和健康对照者（HC组）外周血30 mL，提取并培养EPCs。（1）SDF-1干预组加入100μg/L SDF-1培养液，非干预组加入EGM-2MV培养基，采用Boyden小室和体外血管生成试剂盒观察EPCs的迁移和体外血管生成能力。（2）将培养的EPCs分为空白对照组、1μg/L SDF-1组、10μg/L SDF-1组、100μg/L SDF-1组、单纯AMD3100组及100μg/L SDF-1+AMD3100组，通过Western blot法检测各组EPCs中AKT蛋白的表达水平。结果（1）无SDF-1干预时，DM组EPCs迁移和血管形成能力低于HC组，SDF-1干预后，2组的EPCs迁移和血管形成能力均较干预前增强，但DM组增强的幅度高于HC组。（2）同一浓度下，DM组的AKT蛋白表达水平均低于HC组（均P＜0.01）。无论是DM组还是HC组，AKT蛋白的表达均随着加入SDF-1浓度的增加而呈递增趋势（P＜0.05）；100μg/L SDF-1+AMD3100组AKT蛋白的表达水平较100μg/L SDF-1组明显降低（P＜0.05）。结论 SDF-1可增强外周血EPCs迁移和血管形成能力，对糖尿病患者效果更为明显，且SDF-1对EPCs的影响与PI3K/AKT信号通路有关。
目的：觀察基質細胞衍生因子-1（SDF-1）對糖尿病外週血內皮祖細胞（EPCs）功能的影響，探討SDF-1對EPCs的影響是否與PI3K/AKT信號通路有關。方法採集糖尿病患者（DM組）和健康對照者（HC組）外週血30 mL，提取併培養EPCs。（1）SDF-1榦預組加入100μg/L SDF-1培養液，非榦預組加入EGM-2MV培養基，採用Boyden小室和體外血管生成試劑盒觀察EPCs的遷移和體外血管生成能力。（2）將培養的EPCs分為空白對照組、1μg/L SDF-1組、10μg/L SDF-1組、100μg/L SDF-1組、單純AMD3100組及100μg/L SDF-1+AMD3100組，通過Western blot法檢測各組EPCs中AKT蛋白的錶達水平。結果（1）無SDF-1榦預時，DM組EPCs遷移和血管形成能力低于HC組，SDF-1榦預後，2組的EPCs遷移和血管形成能力均較榦預前增彊，但DM組增彊的幅度高于HC組。（2）同一濃度下，DM組的AKT蛋白錶達水平均低于HC組（均P＜0.01）。無論是DM組還是HC組，AKT蛋白的錶達均隨著加入SDF-1濃度的增加而呈遞增趨勢（P＜0.05）；100μg/L SDF-1+AMD3100組AKT蛋白的錶達水平較100μg/L SDF-1組明顯降低（P＜0.05）。結論 SDF-1可增彊外週血EPCs遷移和血管形成能力，對糖尿病患者效果更為明顯，且SDF-1對EPCs的影響與PI3K/AKT信號通路有關。
목적：관찰기질세포연생인자-1（SDF-1）대당뇨병외주혈내피조세포（EPCs）공능적영향，탐토SDF-1대EPCs적영향시부여PI3K/AKT신호통로유관。방법채집당뇨병환자（DM조）화건강대조자（HC조）외주혈30 mL，제취병배양EPCs。（1）SDF-1간예조가입100μg/L SDF-1배양액，비간예조가입EGM-2MV배양기，채용Boyden소실화체외혈관생성시제합관찰EPCs적천이화체외혈관생성능력。（2）장배양적EPCs분위공백대조조、1μg/L SDF-1조、10μg/L SDF-1조、100μg/L SDF-1조、단순AMD3100조급100μg/L SDF-1+AMD3100조，통과Western blot법검측각조EPCs중AKT단백적표체수평。결과（1）무SDF-1간예시，DM조EPCs천이화혈관형성능력저우HC조，SDF-1간예후，2조적EPCs천이화혈관형성능력균교간예전증강，단DM조증강적폭도고우HC조。（2）동일농도하，DM조적AKT단백표체수평균저우HC조（균P＜0.01）。무론시DM조환시HC조，AKT단백적표체균수착가입SDF-1농도적증가이정체증추세（P＜0.05）；100μg/L SDF-1+AMD3100조AKT단백적표체수평교100μg/L SDF-1조명현강저（P＜0.05）。결론 SDF-1가증강외주혈EPCs천이화혈관형성능력，대당뇨병환자효과경위명현，차SDF-1대EPCs적영향여PI3K/AKT신호통로유관。
Objective To observe the effects of stromal cell-derived-factor-1(SDF-1) on the function of endotheli?al progenitor cells(EPCs)of peripheral blood in patients with diabetes, and to discuss the effects of PI3K/AKT signaling path?way on the role of SDF-1 in EPCs. Methods The peripheral blood samples (30 mL) were collected in 10 diabetes patients (DM group) and 10 healthy controls (HC group). (1) The 100μg/L SDF-1 was added in intervention group. EGM-2MV was added in non-intervention group. The Boyden chamber and in vitro angiogenesis kit were used to analyze the migration and in vitro angiogenesis of EPCs. (2) Cultured EPCs were divided into blank control group, 1μg/L SDF-1 group, 10μg/L SDF-1 group, 100μg/L SDF-1 group, pure AMD3100 group and 100μg/L SDF-1+AMD3100 group. AKT protein expression lev?els of endothelial progenitor cells were detected by Western blot assay in each group. Results (1) Without intervention with SDF-1, EPCs’migration and angiogenesis ability were lower in DM group than those in HC group. After intervention with SDF-1, the migration and angiogenesis ability were enhanced in two groups, but the increased level was higher in DM group than that of HC group. (2) Under the same concentration, AKT protein expression level was significantly lower in DM group than that in HC group (P<0.01). AKT protein expression levels were increased with the increased levels of SDF-1 in DM group and HC group (P<0.05). AKT protein expression was significantly lower in 100μg/L SDF-1+AMD3100 group than that of 100μg/L SDF-1 group (P<0.05). Conclusion SDF-1 can increase the chemotactic migration and angiogenesis ability of EPCs in peripheral blood, especially for patients with diabetes. The effects of SDF-1 on EPCs were related to the PI3K/AKT signaling pathway.
