依达拉奉干预LPS介导原代小胶质细胞的激活实验

乔廷廷; 陈忠义; 董宝莲; 殷燕; 郭玲

依达拉奉干预LPS介导原代小胶质细胞的激活实验

1. 大理大学药学与化学学院
2. 云南省第三人民医院大理大学第二附属医院

基金项目:

基金：国家自然科学基金资助项目 (81760228); 云南省教育厅科学研究基金资助项目 (2012Z093); 云南省科技厅科技计划青年基金资助项目 (2017FD193);

计量
- 文章访问数: 2387
- HTML全文浏览量: 884
- PDF下载量: 77
- 被引次数: 0
出版历程
- 收稿日期: 2018-01-11

The Experiment of Edaravone on Primary Microglia Activation by Lipopolysaccharide

Funds:

基金：国家自然科学基金资助项目 (81760228); 云南省教育厅科学研究基金资助项目 (2012Z093); 云南省科技厅科技计划青年基金资助项目 (2017FD193);

摘要

摘要: 目的探索依达拉奉 (edaravone, EDA) 对脂多糖 (lipopolysaccharide, LPS) 介导原代小胶质细胞 (Microglia) 激活后的一氧化氮 (nitric oxide, NO) 调控作用.方法麻醉SD大鼠新生鼠断头取脑, 消化离心等实验方法完成混合胶质细胞的原代培养, 通过生物摇床方法分离、纯化小胶质细胞.利用Griess法检测一氧化氮释放量.结果通过荧光方法鉴定小胶质细胞的纯化可达到95%以上, 与对照组 (Control) 相比, LPS介导小胶质细胞后激活可见NO浓度显著上升 (P<0.05) , 建模成功;用EDA预处理原代小胶质细胞24 h后, LPS介导的NO下调, 与LPS单独组比较, 差异有统计学意义 (P<0.05) .同时, EDA先预处理24 h, MAPK inhibitor (丝裂原活化蛋白激酶抑制剂) 预处理30 min, 10 ng/m L LPS介导小胶质细胞16 h, NO释放量显著降低, 差异有统计学意义 (P<0.05) .结论 EDA单独应用可抑制LPS介导原代小胶质细胞激活后NO的释放量;EDA同时加用MAPK信号传递通路抑制剂, 加强了EDA减轻LPS介导小胶质细胞激活NO产生.EDA可能与p38MAPK、JNK、ERK信号传导通路抑制剂协同抑制LPS介导的原代小胶质细胞激活有关.
- 依达拉奉 /
- 脂多糖 /
- 小胶质细胞 /
- 一氧化氮
Abstract: Objective To explore the effect of Edaravone (EDA) on oxidative stress such as release of nitric oxide (NO) induced by Lipopolysaccharide (LPS) in activation of primary microglia. Me thods There were four groups including control, LPS, LPS + EDA and MAPK inhibitors + EDA + LPS in this study. The primary mixed glial cells were derived from the cortex of Sprague-Dawley rats which were 2 to 3-day old neonatal pups. The primary microglia were separated and purified by Biological Shaker after growing for about ten days. Griess Assay was used to detect NO secreted by activated microglia in four groups. Re s ults The shake-off microglial cells were confirmed as purity of 95% or up via fluorescence staining of CD68. Compared with the control group, NO product of microglial activation mediated by LPS significantly increased while NO value decreased in the group of EDA which was pretreatedt of EDA ahead of LPS for 24 hrs (P<0.05) . After addition of LPS, the incubation was remained for additional 24 h. In addition to EDA pretreatment for 24 h, the addition of three Mitogen-activated protein kinase inhibitors of MAPK signaling pathways were ahead of addition of EDA for 30 min, and then 10 ng/m Lof LPS was added to microglia. All of them incubated with the primary microglia for 24 h after the last addition. NO release was significantly reduced, the difference was statistically significant (P <0.05) .Conclus ion EDA treatment alone could inhibit the release of NO induced by LPS in primary microglia while EDA plus MAPK signaling pathway inhibitor obviously reduced release of NO. EDA may take effect with p38 MAPK, JNK and ERK inhibitors of sub-pathways in some way (s) to co-operate with EDA in modulating oxidative stress in activated microglia induced by LPS. As to the working mechanisms how EDA co-operates with MAPK inhibitors is still remain unknown.
- Edaravaone /
- Lipopolysaccharide /
- Microglia /
- Nitric oxide

HTML全文

参考文献(27)

[1]章玲, 张衡, 孙秉贵.星形胶质细胞在阿尔茨海默病中的作用及其分子机制[J].2017, 29 (5) :501-506.
[2]沈婕, 李久胜.小胶质细胞在神经退行性疾病中的神经免疫调节作用[J].河南医学研究, 2017, 26 (2) :244-246.
[3]谭子健, 巨淑慧, 黄潇, 等.胶质细胞干细胞/前体细胞特性的研究进展[J].生理学报, 2017, 69 (2) :207-217.
[4]王茜, 程雪娇, 李学敏, 等.小胶质细胞在慢性神经退行性疾病中的作用研究进展[J].卫生研究, 2015, 44 (6) :1023-1027.
[5]蒋宏国.过表达Nurr1基因的小胶质细胞对神经干细胞向多巴胺能神经元分化的影响[D].昆明:昆明医科大学硕士论文, 2015.
[6]	[6]ZHANG P, LI W, LI L, et al.Treatment with edaravone attenuates ischemic brain injury and inhibits neurogenesis in the subventricular zone of adult rats after focal cerebral ischemia and reperfusion injury[J].Neuroscience, 2012, 201 (1) :297-306.
[7]邱奥望, 刘展, 郭军, 等.神经炎症与神经退行性疾病的关系[J].生理科学进展, 2011, 42 (5) :253-258.
[8]	[8]GLASS C K, SAIJO K, WINNER B, et al.Mechanisms underlying inflammation in neurodegeneration[J].Cell, 2010, 140 (6) :918-934.
[9]	[9]GUO L, LJ L D M E.A Dual role for apolipoprotein E in neuroinflammation anti-and pro-inflammatory activity[J], Journal of Molecular Neuroscience, 2004, 23 (3) :205-212.
[10]	[10]GUO L, SAWKAR A, ZASADZKI M, et al.Similar activation of glial cultures from different rat brain regions by neuroinflammatory stimuli and downregulation of the activation by a new class of small molecule ligands[J], Neurobiology ofAging, 2001, 22 (6) :975-981.
[11]	[11]RALAY R H, CRAFT J M, HU W, et al.Glia as a therapeutic target:Selective suppression of human amyloid-β-induced upregulation of brain proinflammatory cytokine production attenuates neurodegeneration[J], The Journal ofNeurobiology, 2006, 26 (2) :662-670.
[12]	[12]WATTERSON DM, MIRZOEVA S, GUO L, et al.Ligand modulation of glial activation:Cell permeable, small molecule inhibitors of serine-threonine protein kinases can block induction of interleukin 1βand nitric oxide synthase II[J].Neurochemistry International, 2001, 39 (5-6) :459-468.
[13]	[13]PETROVA TV, AKAMA KT, LINDA J, et al.Cyclopentenone prostaglandins suppress activation of microglia:Down-regulation of inducible nitric-oxide synthase by15-deoxy-△12, 14-prostaglandin J2[J].Neurobiology, 1999, 96 (8) :4668-4673.
[14]	[14]JINGRU H U, FRANCIS CASTETS, JOSE L.GUEVARA, et al.S100βstimulates inducible nitric oxide synthase activity and m RNA levels in rat cortical astrocytes[J].THE JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (5) :2543-2547.
[15]辛岗, 苏芸, 许燕璇, 等.脂多糖活化小鼠大脑小胶质细胞后炎性细胞因子的表达[J].中国热带医学, 2011, 11 (8) :983-988.
[16]袁龙.脂多糖所介导的小胶质细胞活化的分子机制[D].延吉:延边大学硕士论文, 2012.
[17]	[17]ALOISI F, RIA F, ADORINI L.Regulatiol of T-cell responses by CNS antigen-pres enting cells:different roles for microglia and astrocytes[J].Immunology Today, 2000, 21 (3) :141.
[18] 乔廷廷, 郭玲.MAPK信号传导通路调控Amyloid-beta介导神经炎症中的过氧化物[J].昆明医科大学学报, 2016, 37 (2B) :309-313.
[19]姚惠亚, 张连生, 伍进, 等.依达拉奉对急性脑梗死患者神经元特异性烯醇化酶和C反应蛋白水平及预后的影响[J].中国医师杂志, 2012, 14 (3) :411-413.
[20]袁云, 李璠, 韩宏, 等.依达拉奉通过Notch信号通路调节BV-2小胶质细胞激活的研究[J].神经解剖学杂志, 2017, 33 (5) :560-566.
[21]王漫.JNK抑制剂SP600125对AD脑内突触损害和炎性反应治疗作用及其机制研究[D], 西安:第四军医大学硕士论文, 2015.
[22]	[22]AGTHONG S, KOONAMJ, KAEWSEMA A, et al.Inhibition of MAPK ERK impairs axonal regeneration without an effect on neuronal loss after nerve injury[J].Neurol Res, 2009, 25 (10) :1068-1074.
[23]刘婷婷, 张淑萍, 覃筱燕, 等.MAPK信号传导通路与神经损伤研究进展[J].中国公共卫生, 2016, 32 (2) :248-254.
[24]吴晓, 金茂强, 陈含笑.闭合性弥漫性脑损伤后IL-1β.P38MAPK及VCAM-1的动态表达及意义[J].新疆医科大学学报, 2011, 34 (1) :62-65.
[25]	[25]AVRAMOVICH Y, AMIT T, YOUDIM MB.Non-steroidal anti-inflammatory drugs stimulate secretion of non-amyloidogenic precursor protein[J].The Journal of Biological Chemistry, 2002, 277 (35) :31466-31473.
[26]	[26]WANG S D, CHEN B C, KAO S T, et al.Genistein inhibits tumor invasion by suppressing multiple signal transduction pathways in human hepatocellular carcinoma cells[J].BMCComplementary and Alternative Medicine, 2013, 14 (1) :1-26.
[27]	[27]TSAI J P, HSIAO P C, YANG S F, et al.Licochalcone Asuppresses migration and invasion of human hepatocellular carcinoma cells through downregulation of MKK4/JNK via NF-KB mediated urokinase plasminogen activator exprssion[J].PLoS One, 2014, 9 (1) :e86537.

相关文章(20)

[1]	梁国晶, 冀琨, 张恺纯, 张玉芳, 安晶, 张钰鸽, 文娟, 任海燕. 益生菌对脑缺血再灌注损伤大鼠Aβ表达的影响及神经元的保护作用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240506
[2]	聂绍燕, 范苏苏, 朱钰珊, 彭学容, 王洋, 张旋. 喜炎平注射液对脂多糖诱导的急性肺损伤小鼠的保护作用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20241105
[3]	吕煜, 胡义波, 王嘉鑫, 苏有橦, 代佑果. 蛋白质S-亚硝基化与消化系统肿瘤关系的研究进展, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220911
[4]	赵琨, 肖云, 杨纯, 严志凌, 董敏娜, 向柄全, 肖茗耀. 白细胞介素-4在脂多糖诱导急性肺损伤模型中的保护作用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220805
[5]	朱婷娜, 刘鹏亮, 董文娟, 于浩, 吴亚梅, 黄兆奎, 洪仕君, 赵永娜. 不同剂量天麻素对甲基苯丙胺依赖大鼠条件位置偏爱及海马小胶质细胞激活的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210502
[6]	尹赛格, 张恒睿, 陈滟, 孙俊. 小鼠小胶质细胞原代培养的高产改良方法, 昆明医科大学学报.
[7]	陈忠义, 乔廷廷, 董宝莲, 郭玲. LPS介导三种啮齿动物原代胶质细胞炎症氧化反应的研究, 昆明医科大学学报.
[8]	傅希玥, 陆地, 边立功, 周莉. 小胶质细胞的激活与癫痫的关系, 昆明医科大学学报.
[9]	曹永梅, 吕毅, 刘玉静, 李颖川. miR-200及其靶基因ZEB1/2在急性肺损伤后早期肺纤维化中上皮-间质转化(EMT)过程, 昆明医科大学学报.
[10]	黄胜华, 李晓霞, 连希艳, 陈子摇, 赵劲涛, 冯志坚. 尿酸在氧化应激状态下对人脐静脉内皮细胞的影响, 昆明医科大学学报.
[11]	唐涛, 孙先润, 姜艳萍, 张敏, 马骅, 龙宇. NO/iNOS与关节软骨破坏的相关性, 昆明医科大学学报.
[12]	奎莉越. 降钙素基因相关肽对LPS诱导肺上皮细胞炎症因子的影响, 昆明医科大学学报.
[13]	王慧明. P2X4受体在大鼠脊髓小胶质细胞中的表达, 昆明医科大学学报.
[14]	边海霞. 单纯疱疹病毒性角膜炎抗氧化治疗的临床观察, 昆明医科大学学报.
[15]	王剑. 一重和三重脑震荡大鼠损伤后早期小胶质细胞变化的研究, 昆明医科大学学报.
[16]	姚洁梅. 纳洛酮联合依达拉奉对急性酒精中毒的临床疗效分析, 昆明医科大学学报.
[17]	李福萍. 阿伐他汀对原发性高血压患者硫化氢和一氧化氮水平的影响, 昆明医科大学学报.
[18]	邹智荣. 枫叶黄酮抑制脂多糖诱导破骨前体细胞Raw264.7细胞激活的作用, 昆明医科大学学报.
[19]	. 一重和三重脑震荡大鼠脑小胶质细胞定量变化研究, 昆明医科大学学报.
[20]	唐茂丹. 大鼠视神经切断后视网膜Muller细胞及小胶质细胞变化特点, 昆明医科大学学报.