中山大学学报（医学科学版）
中山大學學報（醫學科學版）
중산대학학보（의학과학판）
JOURNAL OF SUN YAT-SEN UNIVERSITY(MEDICAL SCIENCES)
2010年
1期
64-68
,共5页
谈智%崔雨虹%向秋玲%林平%王庭槐
談智%崔雨虹%嚮鞦玲%林平%王庭槐
담지%최우홍%향추령%림평%왕정괴
雌激素%膜雌激素受体%内皮祖细胞%一氧化氮
雌激素%膜雌激素受體%內皮祖細胞%一氧化氮
자격소%막자격소수체%내피조세포%일양화담
estrogen%membrane estrogen receptor%endothelial progenitor cells%nitric oxide
[目的]观察一氧化氮(NO)信号途径在膜雌激素受体介导的内皮祖细胞(EPCs)增殖和凋亡中的作用.[方法] 在培养的EPCs上,分别使用E_2-BSA、或加雌激素受体阻断剂ICI 182,780、P13K抑制剂LY294002和NOS抑制剂l-NAME,利用噻唑蓝(MTT)法检测细胞增殖活性(10组,n=9),利用化学比色法测定NO的含量(6组,n=6),Hoechst 33258染色观察EPCs凋亡(8组,n=5)以及免疫印迹法检测EPCs磷酸化eNOS蛋白表达的情况(4组,n=4).[结果]与对照组相比,E_2-BSA可以促进EPCs的增殖,ICI 182.780可以抑制其增殖作用,表明膜雌激素受体介导的信号通路参与雌激素对EPCs的增殖作用;NO合成和细胞凋亡结果显示,E_2-BSA可以促进一氧化氮的合成和抑制血清撤退诱导的凋亡,P13K抑制剂LY294002、NOS抑制剂l-NAME以及和ICI 182,780可以抑制上述作用;免疫印迹结果显示,E_2-BSA可以促使eNOS的磷酸化,而LY294002可抑制上述作用.[结论]膜雌激素受体可通过P13K/Akt/eNOS信号途径抑制EPCs的凋亡从而促进其增殖.
[目的]觀察一氧化氮(NO)信號途徑在膜雌激素受體介導的內皮祖細胞(EPCs)增殖和凋亡中的作用.[方法] 在培養的EPCs上,分彆使用E_2-BSA、或加雌激素受體阻斷劑ICI 182,780、P13K抑製劑LY294002和NOS抑製劑l-NAME,利用噻唑藍(MTT)法檢測細胞增殖活性(10組,n=9),利用化學比色法測定NO的含量(6組,n=6),Hoechst 33258染色觀察EPCs凋亡(8組,n=5)以及免疫印跡法檢測EPCs燐痠化eNOS蛋白錶達的情況(4組,n=4).[結果]與對照組相比,E_2-BSA可以促進EPCs的增殖,ICI 182.780可以抑製其增殖作用,錶明膜雌激素受體介導的信號通路參與雌激素對EPCs的增殖作用;NO閤成和細胞凋亡結果顯示,E_2-BSA可以促進一氧化氮的閤成和抑製血清撤退誘導的凋亡,P13K抑製劑LY294002、NOS抑製劑l-NAME以及和ICI 182,780可以抑製上述作用;免疫印跡結果顯示,E_2-BSA可以促使eNOS的燐痠化,而LY294002可抑製上述作用.[結論]膜雌激素受體可通過P13K/Akt/eNOS信號途徑抑製EPCs的凋亡從而促進其增殖.
[목적]관찰일양화담(NO)신호도경재막자격소수체개도적내피조세포(EPCs)증식화조망중적작용.[방법] 재배양적EPCs상,분별사용E_2-BSA、혹가자격소수체조단제ICI 182,780、P13K억제제LY294002화NOS억제제l-NAME,이용새서람(MTT)법검측세포증식활성(10조,n=9),이용화학비색법측정NO적함량(6조,n=6),Hoechst 33258염색관찰EPCs조망(8조,n=5)이급면역인적법검측EPCs린산화eNOS단백표체적정황(4조,n=4).[결과]여대조조상비,E_2-BSA가이촉진EPCs적증식,ICI 182.780가이억제기증식작용,표명막자격소수체개도적신호통로삼여자격소대EPCs적증식작용;NO합성화세포조망결과현시,E_2-BSA가이촉진일양화담적합성화억제혈청철퇴유도적조망,P13K억제제LY294002、NOS억제제l-NAME이급화ICI 182,780가이억제상술작용;면역인적결과현시,E_2-BSA가이촉사eNOS적린산화,이LY294002가억제상술작용.[결론]막자격소수체가통과P13K/Akt/eNOS신호도경억제EPCs적조망종이촉진기증식.
[Objective] The aim of the present study was to investigate the role of membrane estrogen receptor (mER) mediated pathway in the proliferation and apoptosis of endothelial progenitor cells (EPCs). [Methods] Bone marrow (BM)-derived EPCs were cultured. The cells were divided into different groups, plus or not plus estrogen receptor blocker (ICI 182,780), PI3K inhibitors (LY294002), and NOS inhibitor (L-NAME) to show the effect of E_2-BSA on EPCs. The proliferation of EPCs was determined by MTT and nitric oxide (NO) release was measured by chromatometry. Apoptotic cell death was determined using the Hochest 33258 staining. The expression of phosphorylated eNOS (p-eNOS) were detected by Western blot. [Results] E_2-BSA could increase EPCs proliferation, and this effect was inhibited by estrogen receptor blocker ICI 182,780, thus indicated that mER-initiated membrane signaling pathways were involved in the action of estrogen on EPCs. E_2-BSA increased nitric oxide production and inhibited apoptosis induced by serum withdrawal, and this effect also inhibited by PI3K inhibitor (LY294002), NOS inhibitor (L-NAME)and estrogen receptor blocker(ICI 182,780), thus indicated that PI3K/Akt/NO pathway was involved the effect of estrogen on EPCs apoptosis. Moreover, E_2-BSA treatment increased phosphorylation of eNOS (p-eNOS). PI3K inhibitors (LY294002) also blocked these effects. [Conclusions] The results of present study suggested that mER mediated EPCs proliferation and apoptosis were related to the PI3K/Akt/eNOS pathway.
