Hypertonic saline alleviates cerebral edema by inhibiting microglia-derived TNF-α and IL-1β-induced Na-K-Cl Cotransporter up-regulation.
Hypertonic saline alleviates cerebral edema by inhibiting microglia-derived TNF-α and IL-1β-induced Na-K-Cl Cotransporter up-regulation.
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DOI：
10.1186/1742-2094-11-102
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发表时间：
2014-06-11
影响因子：
9.3
通讯作者：
Zeng HK
中科院分区：
文献类型：
--
作者：
Huang LQ;Zhu GF;Deng YY;Jiang WQ;Fang M;Chen CB;Cao W;Wen MY;Han YL;Zeng HK
高渗盐水(HS)已成功地用于临床治疗各种形式的脑水肿。Na-K-Cl协同转运蛋白1及其炎性介质肿瘤坏死因子-α和白介素1-β(IL-1β)的表达上调与多种脑损伤所致脑水肿的发生密切相关。本研究旨在探讨10%HS是否通过下调小胶质细胞炎症介质的表达，从而降低缺血中心区大脑皮质星形胶质细胞NKCC1的表达，从而减轻脑水肿。采用Western blotting、免疫荧光双标记法和实时逆转录聚合酶链式反应(RT-α)检测大鼠大脑中动脉闭塞后NKCC1、肿瘤坏死因子-β和IL-1β的表达，并测定脑含水量和脑梗塞面积。用p38和JNK信号通路的特异性抑制剂SB203580和SP600125处理原代培养的小胶质细胞，用Western blotting、双免疫荧光和酶联免疫吸附试验(ELISA)检测HS对小胶质细胞表达和分泌肿瘤坏死因子-α和IL-1β是否需要这两条信号通路。检测肿瘤坏死因子α和白介素1β对原代培养星形胶质细胞表达NKCC1的影响。此外，还采用Western blotting、免疫荧光双标记法和实时定量RT-PCR检测了HS对原代星形胶质细胞NKCC1表达的直接抑制作用。经10%HS治疗后，BWC和脑梗塞面积明显减少。小胶质细胞内肿瘤坏死因子-α和IL-1β的免疫表达明显降低。同时，星形胶质细胞中NKCC1的表达下调。原代培养的小胶质细胞经低氧暴露和100mM HS处理后，其释放的肿瘤坏死因子-α和IL-1β均减少。原代星形胶质细胞中NKCC1的表达随着外源性肿瘤坏死因子-α和IL-1β浓度的降低而同时和进行性下调。此外，100 mM HS可直接抑制低氧条件下星形胶质细胞NKCC1的上调。结果提示，10%HS通过抑制NKCC1共转运体减轻脑水肿，其机制可能与抑制肿瘤坏死因子α和IL-1β对NKCC1的刺激有关。
Hypertonic saline (HS) has been successfully used clinically for treatment of various forms of cerebral edema. Up-regulated expression of Na-K-Cl Cotransporter 1 (NKCC1) and inflammatory mediators such as tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) has been demonstrated to be closely associated with the pathogenesis of cerebral edema resulting from a variety of brain injuries. This study aimed to explore if alleviation of cerebral edema by 10% HS might be effected through down-regulation of inflammatory mediator expression in the microglia, and thus result in decreased NKCC1 expression in astrocytes in the cerebral cortex bordering the ischemic core. The Sprague-Dawley (SD) rats that underwent right-sided middle cerebral artery occlusion (MCAO) were used for assessment of NKCC1, TNF-α and IL-1β expression using Western blotting, double immunofluorescence and real time RT-PCR, and the model also was used for evaluation of brain water content (BWC) and infarct size. SB203580 and SP600125, specific inhibitors of the p38 and JNK signaling pathways, were used to treat primary microglia cultures to determine whether the two signaling pathways were required for the inhibition of HS on microglia expressing and secreting TNF-α and IL-1β using Western blotting, double immunofluorescence and enzyme-linked immunosorbent assay (ELISA). The effect of TNF-α and IL-1β on NKCC1 expression in primary astrocyte cultures was determined. In addition, the direct inhibitory effect of HS on NKCC1 expression in primary astrocytes was also investigated by Western blotting, double immunofluorescence and real time RT-PCR. BWC and infarct size decreased significantly after 10% HS treatment. TNF-α and IL-1β immunoexpression in microglia was noticeably decreased. Concomitantly, NKCC1 expression in astrocytes was down-regulated. TNF-α and IL-1β released from the primary microglia subjected to hypoxic exposure and treatment with 100 mM HS were decreased. NKCC1 expression in primary astrocytes was concurrently and progressively down-regulated with decreasing concentration of exogenous TNF-α and IL-1β. Additionally, 100 mM HS directly inhibited NKCC1 up-regulation in astrocytes under hypoxic condition. The results suggest that 10% HS alleviates cerebral edema through inhibition of the NKCC1 Cotransporter, which is mediated by attenuation of TNF-α and IL-1β stimulation on NKCC1.
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DOI：
10.1083/jcb.85.3.890
发表时间：
1980-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
McCarthy KD;de Vellis J
通讯作者：
de Vellis J
影响因子：
6.3
作者：
Chen, H;Luo, J;Sun, DD
通讯作者：
Sun, DD
影响因子：
15.9
作者：
Junger, WG;Hoyt, DB;Altman, A
通讯作者：
Altman, A
影响因子：
15.9
作者：
Kreutzberg, GW
通讯作者：
Kreutzberg, GW
DOI：
10.1124/jpet.109.157776
发表时间：
2010-02-01
影响因子：
3.5
作者：
Paulson, Jennifer R.;Yang, Tianzhi;Abbruscato, Thomas J.
通讯作者：
Abbruscato, Thomas J.