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Monocyte-conditioned medium enhances thrombin-stimulated PGI_2 production by human umbilical vein endothelial cells in culture

单核细胞条件培养基增强培养物中人脐静脉内皮细胞凝血酶刺激的 PGI_2 产生

基本信息

批准号：
01570436
负责人：
YOSHIDA Minoru
金额：
$ 1.41万
依托单位：
Fukuoka University, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

1. HUVEC after incubation with LPS (1mug/ml), IL-1alpha (10 U/ml), IL-1beta (10 U/ml), IFN-gamma (1000 U/ml) for 24 hours produced more PGI_2 than the control HUVEC in response to thrombin. Monocyte-conditioned medium was prepared by the incubation of human peripheral blood monocytes in tissue culture flasks with (LPS-Mo-CM) or without LPS (Mo-CM). HUVEC after incubation with Mo-CM produced much more PGI_2 than LPS-, IL-1- or IFN-gamma-treated HUVEC.2. The effect of incubation time in making Mo-CM was examined on thrombin-stimulated PGI_2 production by HUVEC treated with the Mo-CM. The longer was the incubation time in making Mo-CM, the more was PGI_2 production by HUVEC. IL-1beta in Mo-CM was also increased with the incubation time in making Mo-CM.3. HUVEC were cultured for 24 hours with Mo-CM to which excessive-amount of anti-serum for IL-1alpha and/or TNF was added, and were then stimulated with thrombin. Consequent PGI_2 production was decreased, but still much higher than not only the control but also LPS-, IL-1- IFN-gamma-treated HUVEC. There could be produced any other cytokines from monocytes except IL-1 or TNF which stimulate PGI_2 production by HUVEC.4. When HUVEC were cultured with Mo-CM or LPS-Mo-CM, the concentration of IL-1beta in the postculture medium of HUVEC was not significantly changed, compared with Mo-CM or LPS-Mo-CM before HUVEC were cultured with. IL-1beta did not enhance the production of IL-1beta from HUVEC.

1. HUVEC与LPS（1 μ g/ml）、IL-1 α（10 U/ml）、IL-1 β（10 U/ml）、IFN-γ（1000 U/ml）共同孵育24小时后，凝血酶刺激HUVEC产生PGI_2的能力明显高于对照组。单核细胞条件培养基通过将人外周血单核细胞在含有LPS（LPS-Mo-CM）或不含LPS（Mo-CM）的组织培养瓶中孵育来制备。与LPS、IL-1或IFN-γ处理的HUVEC相比，Mo-CM处理的HUVEC产生更多的PGI_2。用凝血酶刺激HUVEC产生PGI_2，观察Mo-CM制备过程中孵育时间对PGI_2产生的影响。培养时间越长，HUVEC产生PGI_2的能力越强。Mo-CM中IL-1 β的含量也随着Mo-CM制备时间的延长而增加。HUVEC与Mo-CM一起培养24小时，Mo-CM中加入过量的IL-1 α和/或TNF的抗血清，然后用凝血酶刺激。结果表明，LPS、IL-1-IFN-γ处理后，HUVEC PGI_2的分泌明显减少，但仍明显高于对照组和LPS、IL-1-IFN-γ处理组。单核细胞除产生IL-1和TNF刺激HUVEC产生PGI_2外，还可产生其它细胞因子. HUVEC与Mo-CM或LPS-Mo-CM共同培养后，培养液中IL-1 β浓度与Mo-CM或LPS-Mo-CM共同培养前相比无明显变化。IL-1 β不促进HUVEC产生IL-1 β。