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中华卫生应急电子杂志 ›› 2018, Vol. 04 ›› Issue (01) : 45 -49. doi: 10.3877/cma.j.issn.2095-9133.2018.01.008

所属专题: 文献

论著

TLR4受体抑制剂对脂多糖诱导的小胶质细胞炎症因子的影响
陈超1, 杨炼红1,()   
  1. 1. 510920 广东广州,中山大学孙逸仙纪念医院神经科
  • 收稿日期:2018-01-30 出版日期:2018-02-18
  • 通信作者: 杨炼红
  • 基金资助:
    广东省自然科学基金(2014A030313085)

Effect of TLR4 inhibitor on the inflammatory cytokines in microglia induced by LPS

Chao Chen1, Lianhong Yang1,()   

  1. 1. Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou 510120, China
  • Received:2018-01-30 Published:2018-02-18
  • Corresponding author: Lianhong Yang
  • About author:
    Corresponding author: Yang Lianhong, Email:
引用本文:

陈超, 杨炼红. TLR4受体抑制剂对脂多糖诱导的小胶质细胞炎症因子的影响[J/OL]. 中华卫生应急电子杂志, 2018, 04(01): 45-49.

Chao Chen, Lianhong Yang. Effect of TLR4 inhibitor on the inflammatory cytokines in microglia induced by LPS[J/OL]. Chinese Journal of Hygiene Rescue(Electronic Edition), 2018, 04(01): 45-49.

目的

探讨TLR4受体抑制剂TAK-242对脂多糖(LPS)诱导的BV2小胶质细胞炎症因子的影响。

方法

用脂多糖诱导BV2细胞构建炎症反应模型。(1)利用CCK-8法检测不同浓度的TAK-242预处理BV2小胶质细胞后的细胞存活率,确定最佳TAK-242浓度。(2)BV2小胶质细胞加入0,0.1,0.5,1μg/mL脂多糖(LPS)刺激24 h后,实时荧光定量PCR(qRT-PCR)法检测TLR4炎症因子mRNA表达的变化。(3)将BV2小胶质细胞分为四组:对照组,TAK-242组,LPS组和TAK-242预处理组。实时荧光定量PCR(qRT-PCR)法检测TLR4、MyD88,IL-1β、IL-6炎症因子mRNA表达的变化。

结果

(1)CCK-8:低剂量的TAK-242不影响BV2细胞活力;与LPS组相比,TAK-242预处理组细胞活力明显升高(P<0.05)。(2)实时荧光定量PCR法:与正常对照组相比,LPS处理组TLR4的mRNA表达明显增加,差异具有统计学意义(P<0.05)。(3)实时荧光定量PCR法:与正常对照组相比,LPS处理组TLR4、MyD88、IL-1β、IL-6的mRNA表达明显增加,差异具有统计学意义(P<0.05);与LPS组相比,TAK-242预处理组TLR4、MyD88、IL-1β、IL-6的mRNA表达明显下降,差异有统计学意义(P<0.05)。

结论

LPS可激活TLR4信号通路;TAK-242可降低疾病因子TLR4、MyD88、IL-1β、IL-6等的mRNA表达,并从而增加细胞存活率。

Objective

To explore the effect of TAK-242, TLR4 inhibitor, on the secreting inflammatory cytokines in BV2 microglia caused by LPS.

Methods

The inflammatory reaction cells were inducedby LPS. (1)Microglias BV2 were cultured with different concentrations of TAK-242, and then , the cell survival rate was detected by cck-8 kit to select the suitable concentration; (2)Microglias BV2 were cultured with different concentrations of lipopolysaccharide(LPS)(0, 0.1, 0.5, 1μg/mL), and then, the TLR4 mRNA expression was detected by Quantitative real-time PCR(qRT-PCR); (3)Microglias BV2 were divided into four groups: Control group, TAK-242 group, LPS group and LPS+ TAK-242 pretreatment group, and then, the TLR4, MYD88, IL-1β, IL-6 mRNA expression were detected by Quantitative real-time PCR(qRT-PCR).

Results

(1)CCK-8: Compared with control group, low concentration of TAK-242 have no effect on the viability of BV2 cells. Compared with LPS group, pretreating of TAK-242(1μM) especially increased the viability of BV2 cells (P<0.05). (2)qRT-PCR: compared with control group, LPS can increase the TLR4 mRNA expression (P<0.05); (3)qRT-PCR: compared with control group, LPS can increase the TLR4mRNA, MyD88mRNA, IL-1βmRNA and IL-6 mRNA expression (P<0.05); Compared with LPS group, pretreating of TAK-242(1μM) can significantly decrease the TLR4mRNA, MyD88mRNA, IL-1βmRNA and IL-6 mRNA expression (P<0.05).

Conclusion

TLR4 signal pathway can be activateed by LPS; TAK-242 can decrease TLR4mRNA, MyD88mRNA, IL-1βmRNA and IL-6 mRNA expression and increase the cells viability.

图1 CCK-8法检测不同浓度的TAK-242对BV2细胞活力的影响
图2 CCK-8法检测TAK-242预处理BV2细胞活力的影响
图3 荧光定量RCR法检测不同浓度的LPS作用于BV2后TLR4mRNA表达情况
图4 荧光定量RCR法检测各组细胞TLR4mRNA表达情况
图5 荧光定量RCR法检测各组细胞MyD88mRNA表达情况
图6 荧光定量RCR法检测各组细胞IL-1βmRNA表达情况
图7 荧光定量RCR法检测各组细胞IL-6mRNA表达情况
1
Vezzani A. Epilepsy and inflammation in the brain: overview and pathophysiology[J]. Epilepsy Curr, 2014, 14(1 Suppl): 3-7.
2
Walker L, Sills GJ.Inflammation and epilepsy: the foundations for a new therapeutic approach in epilepsy?[J]. Epilepsy Curr, 2012, 12(1): 8-12.
3
Kim YS, Joh TH.Microglia, major player in the brain inflammation: their roles in the pathogenesis of Parkinson’s disease[J]. Exp Mol Med, 2006, 38(4): 333-347.
4
Walker DG, Whetzel AM, Lue LF.Expression of suppressor of cytokine signaling genes in human elderly and Alzheimer’s disease brains and human microglia[J]. Neuroscience, 2015, 302: 121-137.
5
Emsley HC, Appleton RE, Whitmore CL, et al. Variations in inflammation-related genes may be associated with childhood febrile seizure susceptibility[J]. Seizure, 2014, 23(6): 457-461.
6
Choy M, Dubé CM, Ehrengruber M, et al.Inflammatory processes, febrile seizures, and subsequent epileptogenesis[J]. Epilepsy Curr, 2014, 14(1 Suppl): 15-22.
7
Matsunaga N, Tsuchimori N, Matsumoto T, et al. TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules[J]. Mol Pharmacol, 2011, 79(1): 34-41.
8
贾建平,陈生弟.神经病学[M].北京:人民卫生出版社,2013:297.
9
Song P, Liu Y, Yu X, et al. Prevalence of epilepsy in China between 1990 and 2015: A systematic review and meta-analysis[J]. J Glob Health, 2017, 7(2): 20706.
10
Nau AL, Mwape KE, Wiefek J, et al.Cognitive impairment and quality of life of people with epilepsy and neurocysticercosis in Zambia[J]. Epilepsy Behav, 2017, S1525-5050(17): 30464-X.
11
Wilcox KS, Vezzani A. Does brain inflammation mediate pathological outcomes in epilepsy?[J]. Adv Exp Med Biol, 2014, 813: 169-183.
12
Devinsky O, Vezzani A, Najjar S, et al.Glia and epilepsy: excitability and inflammation[J]. Trends Neurosci, 2013, 36(3): 174-184.
13
Riazi K, Galic MA, Pittman QJ.Contributions of peripheral inflammation to seizure susceptibility: cytokines and brain excitability[J]. Epilepsy Res, 2010, 89(1): 34-42.
14
Rodgers KM, Hutchinson MR, Northcutt A, et al.The cortical innate immune response increases local neuronal excitability leading to seizures[J]. Brain, 2009, 132(Pt 9): 2478-2486.
15
Klein M, Obermaier B, Angele B, et al.Innate immunity to pneumococcal infection of the central nervous system depends on toll-like receptor (TLR) 2 and TLR4[J]. J Infect Dis, 2008, 198(7): 1028-1036.
16
Medzhitov R, Preston-Hurlburt P, Janeway C J. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J]. Nature, 1997, 388(6640): 394-397.
17
Wu Y, Li W, Zhou C, et al.Ketamine inhibits lipopolysaccharide-induced astrocytes activation by suppressing TLR4/NF-kB pathway[J]. Cell Physiol Biochem, 2012, 30(3): 609-617.
18
Cohen P. The TLR and IL-1 signalling network at a glance[J]. J Cell Sci, 2014, 127(Pt 11): 2383-2390.
19
Akira S, Uematsu S, Takeuchi O. Pathogen recognition and innate immunity[J]. Cell, 2006, 124(4): 783-801.
20
Rice TW, Wheeler AP, Bernard GR, et al.A randomized, double-blind, placebo-controlled trial of TAK-242 for the treatment of severe sepsis[J]. Crit Care Med, 2010, 38(8): 1685-1694.
21
Rajamani U, Jialal I. Hyperglycemia induces Toll-like receptor-2 and -4 expression and activity in human microvascular retinal endothelial cells: implications for diabetic retinopathy[J]. J Diabetes Res, 2014, 2014: 790902.
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