Inducible nitric oxide synthase is key to peroxynitrite-mediated, LPS-induced protein radical formation in murine microglial BV2 cells.
Inducible nitric oxide synthase is key to peroxynitrite-mediated, LPS-induced protein radical formation in murine microglial BV2 cells.
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DOI：
10.1016/j.freeradbiomed.2014.04.014
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发表时间：
2014-08
影响因子：
7.4
通讯作者：
Mason, Ronald P.
中科院分区：
文献类型：
--
作者：
Kumar, Ashutosh;Chen, Shih-Heng;Kadiiska, Maria B.;Hong, Jau-Shyong;Zielonka, Jacek;Kalyanaraman, Balaraman;Mason, Ronald P.
关键词：
小胶质细胞是大脑中的常驻免疫细胞。小胶质细胞活化是几种炎性和神经退行性疾病的特征，包括阿尔茨海默病、多发性硬化和帕金森病。虽然在帕金森病（PD）模型中LPS诱导的小胶质细胞活化被充分记录，但在LPS诱导的小胶质细胞活化期间自由基介导的蛋白质自由基形成及其潜在机制尚不清楚。在这里，我们已经使用免疫自旋捕获和RNA干扰，研究诱导型一氧化氮合酶（iNOS）的作用，过氧亚硝基介导的蛋白质自由基的形成在小鼠小胶质细胞BV 2细胞与LPS处理。用LPS处理BV 2细胞导致形态学改变，诱导型一氧化氮合酶的诱导和蛋白自由基的形成增加。FeTPPS、L-NAME、1400 W和夹竹桃素预处理可明显抑制LPS诱导的蛋白质自由基形成和酪氨酸硝化。香豆素-7-硼酸，一个高度特异性的探针过氧亚硝酸盐检测，与LPS诱导的酪氨酸硝化，这表明参与蛋白质自由基的形成过氧亚硝酸盐得到的结果。1400 W和L-NAME所赋予的类似程度的保护使我们得出结论，只有iNOS，而没有其他形式的NOS，参与LPS诱导的过氧亚硝酸盐形成。随后，使用针对iNOS的siRNA、iNOS特异性抑制剂1400 W、NF-kB抑制剂PDTC和P38 MAPK抑制剂SB 202190直接或间接抑制iNOS。在LPS处理的BV 2细胞中，iNOS的抑制与蛋白自由基形成的减少精确相关。蛋白自由基形成的时间过程也与iNOS表达的时间过程相匹配。总之，这些结果证明了iNOS在LPS处理的小胶质细胞BV 2细胞中过氧亚硝酸盐介导的蛋白质自由基形成中的作用。
Microglia are the resident immune cells in the brain. Microglial activation is characteristic of several inflammatory and neurodegenerative diseases including Alzheimer’s disease, multiple sclerosis, and Parkinson’s disease. Though LPS-induced microglial activation in models of Parkinson’s disease (PD) is well documented, the free radical-mediated protein radical formation and its underlying mechanism during LPS-induced microglial activation is not known. Here we have used immuno-spin trapping and RNA interference to investigate the role of inducible nitric oxide synthase (iNOS) in peroxynitrite-mediated protein radical formation in murine microglial BV2 cells treated with LPS. Treatment of BV2 cells with LPS resulted in morphological changes, induction of iNOS and increased protein radical formation. Pretreatments with FeTPPS (a peroxynitrite decomposition catalyst), L-NAME (total NOS inhibitor), 1400W (iNOS inhibitor) and apocynin significantly attenuated LPS-induced protein radical formation and tyrosine nitration. Results obtained with coumarin-7-boronic acid, a highly specific probe for peroxynitrite detection, correlated with LPS-induced tyrosine nitration, which demonstrated involvement of peroxynitrite in protein radical formation. A similar degree of protection conferred by 1400W and L-NAME led us to conclude that only iNOS, and no other forms of NOS, are involved in LPS-induced peroxynitrite formation. Subsequently, siRNA for iNOS, the iNOS-specific inhibitor 1400W, the NF-kB inhibitor PDTC and the P38 MAPK inhibitor SB202190 were used to inhibit iNOS directly or indirectly. Inhibition of iNOS precisely correlated with decreased protein radical formation in LPS-treated BV2 cells. The time course of protein radical formation also matched the time course of iNOS expression. Taken together, these results prove the role of iNOS in peroxynitrite-mediated protein radical formation in LPS-treated microglial BV2 cells.
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影响因子：
15.9
作者：
Kreutzberg, GW
通讯作者：
Kreutzberg, GW
影响因子：
6.7
作者：
Han, Jung Eun;Choi, Ji Woong
通讯作者：
Choi, Ji Woong
影响因子：
2.9
作者：
He, Ya-fang;Chen, Hui-jin;Buzby, Jeffrey S.
通讯作者：
Buzby, Jeffrey S.
DOI：
10.1111/j.1440-1681.2012.05716.x
发表时间：
2012-07-01
影响因子：
2.9
作者：
Kaneko, Yoko S.;Ota, Akira;Nagatsu, Toshiharu
通讯作者：
Nagatsu, Toshiharu
影响因子：
3.3
作者：
BLASI, E;BARLUZZI, R;BISTONI, F
通讯作者：
BISTONI, F