多巴胺D1类受体在小鼠巨噬细胞上的表达及其对ox.docVIP

多巴胺D1类受体在小鼠巨噬细胞上的表达及其对ox-LDL诱导的小鼠巨噬细胞增殖的影响 杨迪，韩 愈，寇 恂，符金娟，刘渔凯，王 震，何多芬，曾春雨 （400042 重庆，第三军医大学大坪医院心血管病专科医院心内科、重庆市心血管病研究所） 摘要：目的 检测小鼠巨噬细胞上的多巴胺D1类表达及探讨其对ox-LDL诱导的小鼠巨噬细胞增殖的影响。方法 以小鼠巨噬细胞RAW264.7为靶细胞，Western blot法检测多巴胺D1类受体的表达。不同浓度的ox-LDL（10、20、40ug/ml）诱导RAW264.7细胞的增殖，观察在多巴胺受体激动剂（Fenoldopam）10-7mol./L、拮抗剂（SCH23390）10-6mol./L及MAPK信号通路的特异性阻断剂（PD98059）10-7mol./L存在的情况下，RAW264.7增殖的变化情况，细胞增殖用3H- TdR掺入量表示。Western blot法检测磷酸化的ERK、总ERK1/2及增殖细胞核抗原PCNA的表达。结果 多巴胺D1类受体在RAW264.7细胞上表达。Fenoldopam（10-7mol./L）、PD98059（10-6mol./L）能明显抑制ox-LDL诱导的RAW264.7细胞增殖，且SCH23390预处理可以逆转Fenoldopam对RAW264.7细胞增殖的抑制作用。在MAPK 阻断剂PD98059存在的情况下，Fenoldopam作用丧失，此外，Fenoldopam（10-7mol./L）可降低ox-LDL升高的PCNA以及磷酸化ERK水平。结论 小鼠巨噬细胞RAW264.7上有多巴胺D1类受体的表达，且刺激多巴胺D1类受体可明显抑制ox-LDL诱导的RAW264.7细胞增殖作用，该作用可能通过影响MAPK/ERK这一信号转导通路来实现。 关键词：多巴胺D1类受体；巨噬细胞增殖；动脉粥样硬化 Expression and Role of Dopamine D1-like Receptors in ox-LDL-induced Proliferation of Macrophages Abstract：Objective To investigate the expression of dopamine D1-like receptors in RAW264.7 mouse macrophages and role of the receptors in ox-LDL-induced proliferation of macrophages..Methods Mouse macrophages RAW264.7 were used as target cells,Western blotting investigate the expression of dopamine D1-like receptors in RAW264.7 mouse macrophages.The RAW264.7 mouse macrophages were treated by ox-LDL(20、40、60ug/ml) with or without the presence of Fenoldopam,PD98059（10-7mol/L）and SCH23390(10-6mol./L). The proliferation of RAW264.7 mouse macrophages was determined by 3H- TdR incorporation and Western blotting investigate the expression of p-ERK and PCNA.Results RAW264.7 cell express D1-like Receptors. Fenoldopam（10-7mol./L）、PD98059（10-6mol/L）can Significantly inhibit the ox-LDL-induced proliferation of macrophages,and SCH23390 blocked the stimulatory effect of Fenoldpam on RAW264.7 cell proliferation.In addition,Fenoldpam decreased the expression of PCNA and p- ERK.Conclusion These data indicate that macrophage express.. D1-like Receptors. Activation of D1-like Receptors r