中华胸心血管外科杂志
中華胸心血管外科雜誌
중화흉심혈관외과잡지
Chinese Journal of Thoracic and Cardiovascular Surgery
2010年
2期
116-119
,共4页
闫醒军%边忠平%刘勇%黄启荣%崔驰%周秀娟%杨辉%钟武%曾宏%施森%姜隽%何延政
閆醒軍%邊忠平%劉勇%黃啟榮%崔馳%週秀娟%楊輝%鐘武%曾宏%施森%薑雋%何延政
염성군%변충평%류용%황계영%최치%주수연%양휘%종무%증굉%시삼%강준%하연정
糖基化产物,高级%内皮,血管%细胞凋亡
糖基化產物,高級%內皮,血管%細胞凋亡
당기화산물,고급%내피,혈관%세포조망
Glycation end products,advanced%Endothelium,vascular%Apoptosis
目的 观察糖基化终末产物(AGEs)对体外培养的人外周血内皮祖细胞(EPCs)生物学特性的影响.方法 以密度梯度离心法获取人外周血单个核细胞(MNCs),由激光共聚焦显微镜鉴定FITC-UEA-1和Dil-acLDL双染色阳性细胞为正在分化的EPCs.加入不同浓度AGEs培养48h,然后分别采用MTT法、Boyden小室测定来观察EPCs的增殖、迁移能力;以人纤维连接蛋白(hFN)检测EPCs的黏附能力;分别采用甲醛和Dnase Ⅰ诱导EPCs凋亡作为阳性对照组,用AnnexinV-FITC/PI和TUNEL法流式细胞仪检测AGEs对EPCs凋亡率的影响.结果 高浓度AGEs可减少EPCs贴壁细胞数量(P<0.01),明显减弱EPCs的黏附(P<0.01)、增殖(P<0.001)、迁移能力(P<0.05),提高EPCs的早期凋亡率(P<0.001).结论 AGEs可减少EPCs的数昔并使EPCs的功能受损.
目的 觀察糖基化終末產物(AGEs)對體外培養的人外週血內皮祖細胞(EPCs)生物學特性的影響.方法 以密度梯度離心法穫取人外週血單箇覈細胞(MNCs),由激光共聚焦顯微鏡鑒定FITC-UEA-1和Dil-acLDL雙染色暘性細胞為正在分化的EPCs.加入不同濃度AGEs培養48h,然後分彆採用MTT法、Boyden小室測定來觀察EPCs的增殖、遷移能力;以人纖維連接蛋白(hFN)檢測EPCs的黏附能力;分彆採用甲醛和Dnase Ⅰ誘導EPCs凋亡作為暘性對照組,用AnnexinV-FITC/PI和TUNEL法流式細胞儀檢測AGEs對EPCs凋亡率的影響.結果 高濃度AGEs可減少EPCs貼壁細胞數量(P<0.01),明顯減弱EPCs的黏附(P<0.01)、增殖(P<0.001)、遷移能力(P<0.05),提高EPCs的早期凋亡率(P<0.001).結論 AGEs可減少EPCs的數昔併使EPCs的功能受損.
목적 관찰당기화종말산물(AGEs)대체외배양적인외주혈내피조세포(EPCs)생물학특성적영향.방법 이밀도제도리심법획취인외주혈단개핵세포(MNCs),유격광공취초현미경감정FITC-UEA-1화Dil-acLDL쌍염색양성세포위정재분화적EPCs.가입불동농도AGEs배양48h,연후분별채용MTT법、Boyden소실측정래관찰EPCs적증식、천이능력;이인섬유련접단백(hFN)검측EPCs적점부능력;분별채용갑철화Dnase Ⅰ유도EPCs조망작위양성대조조,용AnnexinV-FITC/PI화TUNEL법류식세포의검측AGEs대EPCs조망솔적영향.결과 고농도AGEs가감소EPCs첩벽세포수량(P<0.01),명현감약EPCs적점부(P<0.01)、증식(P<0.001)、천이능력(P<0.05),제고EPCs적조기조망솔(P<0.001).결론 AGEs가감소EPCs적수석병사EPCs적공능수손.
Objective In addition to be involved in the angiogenesis, endothelial progenitor cells (EPCs) have roles in endothelium repairing, wound healing, and for protecting blood vessels from restenosis, Advanced glycation end products (AGEs) facilitate the development and progression of atherosclerosis, diabetes associated vascular complications and uremia through various mechanisms such as damaging the endothelium, promoting leukocyte adhension, increasing the aggregation of platelets, and stimulating the proliferation of vascular smooth muscles. This study was designed to explore whether AGEs have effects on biological characteristics of EPCs in cultured human peripheral blood cells. Methods Total mononuclear cells (MNCs), isolated from human peripheral blood by density gradient centrifugatian and adherence cells filtration, were incuba-ted in fibronectin-coated culture dishes. Endothelial cells were identified by means of the adsorption of ulex eurepaeus-aggluti-nin- Ⅰ (UEA- Ⅰ) labelled with fluorescein isothiacyanate (FITC) and Dil-acLDL internalization. Four days later,various con-centrations of AGEs were added to the adherent cells and remained for48 hours. MTT assay and Boyden chamber were used for observing the proliferation and migration of EPCs. Human fibronectin was used to examine the adhesion ability of EPCs. Apop-tosis was induced in the EPCs with formaldehyde and Dnase Ⅰ as a positive control group. Annexin V-FITC/PI and TUNEL method of flow cytometry were used for evaluating the effects of AGEs on the rate of apeptosis in the EPCs. Results AGEs at high concentration decreased the number of EPCs independently (P < 0.01) ; reduced the proliferation (P < 0.01), migration (P<0.001) and adhesive capacity (P<0.05) of EPCs significantly,as well as increasing the apoptasis rate of EPCs in the early stage (P < 0.001). Conclusion AGEs may have adverse effects on EPCs from cultured human peripheral MNCs, such as decreasing their numbers and impairing their functions.
