Inhibition of endotoxin‐induced nitric oxide synthase production in microglial cells by the presence of astroglial cells: A role for transforming growth factor β
Inhibition of endotoxin‐induced nitric oxide synthase production in microglial cells by the presence of astroglial cells: A role for transforming growth factor β
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星形胶质细胞的存在抑制小胶质细胞中内毒素诱导的一氧化氮合酶的产生：转化生长因子β的作用
DOI：
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
A. van Dam
中科院分区：
文献类型：
--
作者：
V. Vincent;F. Tilders;A. van Dam
在新生大鼠大脑皮质的混合胶质细胞培养物中，内毒素诱导小胶质细胞产生诱导型一氧化氮（iNOS）、一氧化氮（NO）和白细胞介素-1 β（IL-1β）。早期我们证明了内毒素诱导的小胶质细胞中iNOS而不是IL-1β的表达被星形胶质细胞的存在抑制。在本文中，我们描述了星形胶质细胞对小胶质细胞表达iNOS的选择性抑制作用的机制的研究。通过单标记或双标记免疫细胞化学技术研究iNOS和IL-1β的表达，并用GSA-I-B4-同工凝集素和抗GFAP抗体进行细胞鉴定。通过分别测量细胞裂解物和培养基中的IL-1β和亚硝酸盐浓度来确定IL-1β和NO的产生。使用生物测定法在混合胶质细胞培养物的培养基中测量了TGFβ（一种已知可抑制内毒素攻击的巨噬细胞产生NO的细胞因子）。小胶质细胞、星形胶质细胞和混合胶质细胞培养物产生相似浓度的TGFβ。应用免疫中和抗体检测TGFβ1和TGFβ2对星形胶质细胞诱导小胶质细胞iNOS表达的影响。用这些TGFβ抗体（3 μg/ml）孵育混合胶质细胞培养物显著增加了小胶质细胞中内毒素诱导的NO产生和iNOS表达，而IL-1β的产生不受影响。抗TGFβ1和TGFβ2的抗体在纯的小胶质细胞培养物中略微增加NO的产生，然而在纯化的小胶质细胞培养物中，重组TGFβ1和TGFβ2与内毒素一起抑制NO的产生。我们的结论是，星形胶质细胞的存在是必要的抑制作用的TGFβ对小胶质细胞的NO产生（可能）通过激活TGFβ或通过增加小胶质细胞的敏感性TGFβ。GLIA 19：190-198，1997.© 1997 Wiley利斯公司
In mixed glial cell cultures from cerebral cortices of newborn rats, endotoxin induces inducible nitric oxide (iNOS), nitric oxide (NO), and interleukin‐1β (IL‐1β) production in microglial cells. Earlier we demonstrated that endotoxin induced iNOS but not IL‐1β expression in microglial cells is inhibited by the presence of astroglial cells. In the present paper we describe studies on the mechanism by which astroglial cells exert selective suppressive action on iNOS expression by microglial cells. Expression of iNOS and IL‐1β was studied by single or double label immunocytochemical techniques and cell identification was performed with GSA‐I‐B4‐isolectin and an antibody against GFAP. Production of IL‐1β and NO was determined by measurement of IL‐1β and nitrite concentrations in cell lysates and the culture medium, respectively. TGFβ, a cytokine known to inhibit NO production by endotoxin challenged macrophages, was measured in culture medium of mixed glial cell cultures using a bioassay. Microglial, astroglial, and mixed glial cell cultures produced similar concentrations of TGFβ. The potential effect of TGFβ was studied by using immunoneutralizing antibodies against TGFβ1 and TGFβ2 on the induction of iNOS in microglial cells in the presence of astroglial cells. Incubation of the mixed glial cell culture with these TGFβ antibodies (3 μg/ml) markedly increased endotoxin‐induced NO production and iNOS expression in microglial cells, whereas the production of IL‐1β was not affected. The antibodies against TGFβ1 and TGFβ2 marginally increased NO production in pure microglial cell cultures, nonetheless in cultures of purified microglial cells recombinant TGFβ1 and TGFβ2 together with endotoxin inhibited NO production. We conclude that the presence of astroglial cells is essential for the inhibitory effect of TGFβ on NO production by microglial cells (possibly) by activation of TGFβ or by increasing the sensitivity of microglial cells for TGFβ. GLIA 19:190–198, 1997. © 1997 Wiley‐Liss, Inc.
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发表时间：
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期刊：
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DOI：
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发表时间：
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期刊：
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影响因子：
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