摘要
目的探讨KCa3.1通道对内皮祖细胞(endothelial progenitor cell,EPCs)生物学功能及分化的影响。方法应用qRTPCR检测EPCs上KCa3.1、vWF、CD31基因表达;采用CCK-8法、细胞贴壁法及Matrigel基质胶分别检测细胞增殖、黏附及体外血管形成能力的变化;应用细胞免疫荧光、Western blot及流式细胞术分别检测KCa3.1通道蛋白、内皮分化标志v WF和CD31、整合素β_1、β_3的表达。结果阻断KCa3.1功能或干扰KCa3.1的表达能够降低EPCs的增殖、黏附及血管形成能力,但是却可以促进EPCs向内皮分化;激活或过表达KCa3.1通道能够增强EPCs的增殖、黏附及血管形成能力,但是阻碍EPCs向内皮分化。阻断KCa3.1离子通道后,细胞膜整合素β_1表达下调,而激活KCa3.1离子通道整合素β_1表达上调。结论 KCa3.1通道功能变化及表达变化能够改变EPCs的生物学特性及向内皮分化水平。
OBJECTIVE To investigate the effect of KCa3. 1 channel on the function of EPCs. METHODS The gene expression of KCa3. 1,v WF and CD31 on EPCs were detected by qRT-PCR. CCK-8 kit,cell adherent method and Matrigel were used to detect the changes of cell proliferation,adhesion and in vitro angiogenesis; cell immunofluorescence or fluorescence activated cell sorter( FACS) was used to detect the protein expression of KCa3. 1,v WF,CD31,integrin β1,integrin β3 separately. RESULTS Blocking the function of KCa3. 1 or interfering with the expression of KCa3. 1 can attenuated EPC function of proliferation,adhesion and angiogenesis,but it can promote the differentiation of EPCs. Overexpression or activation of KCa3. 1 channel can enhance EPCs proliferation,adhesion and angiogenesis but decrease the level of differentiation. The expression of integrin β1 on EPCs was attenuated with blocking KCa3. 1 channel,but the expression effect was reversible by the activator. CONCLUSION The alteration of KCa3. 1 channel function or expression affect the biological characteristics and differentiation of EPCs.
引文
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