血管紧张素Ⅱ对人单核细胞CD40信号系统的作用研究

Effect of angiotensinⅡ on CD40 signaling system in human monocyte cells

目的：以体外培养的单核细胞系为研究对象,从细胞水平探讨血管紧张素Ⅱ对CD40信号系统的影响。方法：用1640培养液进行单核细胞的培养,并用佛波醇肉豆蔻乙酸酯（PMA）诱导其分化为巨噬细胞。将培养的细胞置于不同浓度的血管紧张素Ⅱ（10-10~10-5mol/L）及拮抗剂氯沙坦、PD123319中培养24 h,用实时聚合酶链反应（PCR）的技术检测细胞CD40mRNA的变化;同时酶联免疫法测定细胞上清分泌的CD40蛋白水平。结果：给予PMA（50ng/ml）刺激4 h后,单核细胞贴壁分化为巨噬细胞,与不同浓度血管紧张素Ⅱ（10-10~10-5mol/L）共培养24 h后,与对照组相比较,血管紧张素Ⅱ的10-6mol/L组与10-5mol/L组CD40 mRNA的表达均明显增加（P均〈0.05）;受体AT1拮抗剂氯沙坦（25μg/ml）抑制了CD40 mRNA的表达,另一受体AT2拮抗剂PD123319（25μg/ml）显示了明显的相反作用（P〈0.05）。含单核细胞分化成的巨噬细胞上清液中CD40蛋白水平在血管紧张素Ⅱ及其受体拮抗剂的刺激下显示与CD40 mRNA水平一致的结果。结论：血管紧张素Ⅱ对于单核细胞转化生成的巨噬细胞CD40-CD40L系统的表达具有上调作用。血管紧张素Ⅱ两种受体拮抗剂对CD40系统的作用不同,提示肾素-血管紧张素系统与CD40信号系统之间具有一定相互作用。

Objective：To investigate the influence of angiotensin Ⅱ（AngⅡ） on CD40 signaling system in THP-1 monocyte cell line and macrophages.Methods：THP-1 monocyte cells were cultured in 1640 medium and induced to macrophages by phorbol myristate acetate（PMA）.Then the macrophages were cultured in medium containing AngⅡ（10-10～10-5mol/L） and antagonist losartan,PD123319 for 24h.CD40 mRNA level was analyzed by real-time Taqman polymerase chain reaction（PCR）.And supernatant CD40 level was examined by enzyme-linked immunosorbent assay.Results：THP-1 monocyte cells were induced to macrophages by PMA（50ng/ml） after 4h.After exposed to varied concentrations of AngⅡ（10-10～10-5mol/L）for 24h,expressions of CD40 mRNA in AngⅡ10-6mol/L group and 10-5mol/L group significantly increased compared with control group（P0.05 both）;AT1 antagonist losartan（25μg/ml） inhibited the expression of CD40 mRNA whereas AT2 antagonist PD123319（25μg/ml） showed significant opposite effect（P0.05）.After exposed to AngⅡ and its antagonists,CD40 protein levels of supernatant of macrophages showed results parallel to those of CD40 mRNA levels.Conclusion：Angiotensin Ⅱ enhanced the expression of CD40 signalling system in macrophages;two kinds of AT receptor antagonists showed opposite effects on CD40 system.It indicates some relationship between renin-angiotensin-system and CD40 signalling system.