介入放射学杂志
介入放射學雜誌
개입방사학잡지
JOURNAL OF INTERVENTIONAL RADIOLOGY
2014年
6期
511-515
,共5页
魏黎明%付银鑫%朱悦琦%鲁海涛%王珏%程永德%李晓聪%汪泱%赵俊功
魏黎明%付銀鑫%硃悅琦%魯海濤%王玨%程永德%李曉聰%汪泱%趙俊功
위려명%부은흠%주열기%로해도%왕각%정영덕%리효총%왕앙%조준공
糖尿病%内皮祖细胞%生物活性
糖尿病%內皮祖細胞%生物活性
당뇨병%내피조세포%생물활성
diabetic mellitus%endothelial progenitor cells%biological activity
目的:探讨糖尿病大鼠骨髓来源内皮祖细胞与正常大鼠在数量及生物学活性方面的差异。方法雄性SD大鼠按60 mg/kg腹腔快速注射1%STZ的枸椽酸缓冲液建立糖尿病动物模型。密度梯度法分离糖尿病大鼠和正常大鼠骨髓单个核细胞,用EGMTM-2MV BulletKitTM培养基进行诱导培养,观察细胞形态学特征,流式细胞仪检测细胞表面抗原,摄取Dil-Ac-LDL 与结合 FITC-UEA-1双荧光鉴定细胞及检测细胞比例,CCK-8、Transwell小盒等检测细胞增殖、黏附及迁移能力。结果内皮祖细胞在体外培养时呈梭形,贴壁生长,均可表达CD34、CD133和VEGFR-2,且能摄取Dil-Ac-LDL并结合FITC-UEA-1,证明所培养细胞为内皮祖细胞。糖尿病大鼠骨髓来源的内皮祖细胞数目与正常大鼠相比无明显差异,但其黏附能力明显低于正常大鼠,增殖能力、黏附及迁移能力均降低。结论与正常大鼠相比,糖尿病大鼠骨髓来源内皮祖细胞数量无明显差异,但生物学活性降低,主要表现在增殖能力、黏附及迁移能力均减弱。
目的:探討糖尿病大鼠骨髓來源內皮祖細胞與正常大鼠在數量及生物學活性方麵的差異。方法雄性SD大鼠按60 mg/kg腹腔快速註射1%STZ的枸椽痠緩遲液建立糖尿病動物模型。密度梯度法分離糖尿病大鼠和正常大鼠骨髓單箇覈細胞,用EGMTM-2MV BulletKitTM培養基進行誘導培養,觀察細胞形態學特徵,流式細胞儀檢測細胞錶麵抗原,攝取Dil-Ac-LDL 與結閤 FITC-UEA-1雙熒光鑒定細胞及檢測細胞比例,CCK-8、Transwell小盒等檢測細胞增殖、黏附及遷移能力。結果內皮祖細胞在體外培養時呈梭形,貼壁生長,均可錶達CD34、CD133和VEGFR-2,且能攝取Dil-Ac-LDL併結閤FITC-UEA-1,證明所培養細胞為內皮祖細胞。糖尿病大鼠骨髓來源的內皮祖細胞數目與正常大鼠相比無明顯差異,但其黏附能力明顯低于正常大鼠,增殖能力、黏附及遷移能力均降低。結論與正常大鼠相比,糖尿病大鼠骨髓來源內皮祖細胞數量無明顯差異,但生物學活性降低,主要錶現在增殖能力、黏附及遷移能力均減弱。
목적:탐토당뇨병대서골수래원내피조세포여정상대서재수량급생물학활성방면적차이。방법웅성SD대서안60 mg/kg복강쾌속주사1%STZ적구연산완충액건립당뇨병동물모형。밀도제도법분리당뇨병대서화정상대서골수단개핵세포,용EGMTM-2MV BulletKitTM배양기진행유도배양,관찰세포형태학특정,류식세포의검측세포표면항원,섭취Dil-Ac-LDL 여결합 FITC-UEA-1쌍형광감정세포급검측세포비례,CCK-8、Transwell소합등검측세포증식、점부급천이능력。결과내피조세포재체외배양시정사형,첩벽생장,균가표체CD34、CD133화VEGFR-2,차능섭취Dil-Ac-LDL병결합FITC-UEA-1,증명소배양세포위내피조세포。당뇨병대서골수래원적내피조세포수목여정상대서상비무명현차이,단기점부능력명현저우정상대서,증식능력、점부급천이능력균강저。결론여정상대서상비,당뇨병대서골수래원내피조세포수량무명현차이,단생물학활성강저,주요표현재증식능력、점부급천이능력균감약。
Objective To study the defects of bone marrow-derived endothelial progenitor cells (EPCs) in number ratio and biological abilities (proliferation, adhesion and migration) in diabetic rats. Methods (1) Establishment of diabetic rat model:1%STZ solution was quickly injected into the abdominal cavity of the male SD rats with the dose of 60 mg/kg. (2). Isolation, culture and identification of bone marrow-derived EPCs in diabetic and normal rats. Bone marrow mononuclear cells were isolated from diabetic and normal rats by density gradient centrifugation methods and cultured by EGM-2 MV medium. The cells were identified by morphological observation, FITC-UEA-1 binding and Dil-Ac-LDL uptake assay, and fluorescent immunocytochemistry was used for detection of CD34 , CD133 and VEGFR-2 expression. CCK-8 method and Transwell kit method were used to determine biological activities of EPCs. Results (1) When cultured in vitro, both bone marrow-derived EPCs in diabetic and normal rats were fusiform in shape, the cells snuggled up to the wall. The expression of CD34, CDl33, VEGFR-2 could be detected in these cells, and the cells could uptake Dil-Ac-LDL and bind FITC-UEA-1, which proved that these cells were EPCs. (2) No significant difference in the number of EPCs derived from bone marrow existed between diabetic rats and normal rats, but the proliferation ability, migration ability and adhesion ability of bone marrow-derived EPCs in diabetic rats were obviously lower than those in normal rats. Conclusion The number of bone marrow-derived EPCs in diabetic rats is not obviously different from that in normal rats, but the biologic activity of EPCs derived from bone marrow in diabetic rats is degraded, which is manifested as weakened abilities of the proliferation, adhesion and migration.