TLR4/MyD88介导的炎症反应在体外培养癫痫海马组织中的作用
摘要
目的研究在无镁液所致自发性癫痫样放电海马组织中Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(myeloid differentiation factor 88,My D88)、IL-1β和IL-6的表达,探讨TLR4介导的炎症反应在癫痫发病中的作用。方法取新生6～9 d SD鼠的海马,体外培养至7 d,随机分为正常对照组、癫痫样放电脑片模型组(无Mg~(2+)ACSF灌流3 h),于造模前和造模后1 d、3 d、7 d,采用qRT-PCR检测TLR4、MyD88、IL-1β、IL-6基因,采用Western blot检测TLR4和MyD88蛋白的表达。结果与体外培养条件下正常海马组织相比,无镁细胞外液灌流后的癫痫样放电模型的TLR4、MyD88、IL-1β、IL-6 mRNA升高,TLR4和MyD88的蛋白表达升高,并且随时间延长而增强(P<0.001)。结果体外培养无Mg~(2+)ACSF所致的癫痫样放电海马组织中,可通过TLR4-MyD88信号通路,引起炎症因子IL-1β、IL-6增多,在癫痫的发病机制中可能发挥一定的作用。
引文
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[2]DO NASCIMENTO AL,DOS SANTOS NF,CAMPOS PELáGIOF,et al.Neuronal degeneration and gliosis time-course in the mouse hippocampal formation after pilocarpine-induced status epilepticus[J].Brain Research,2012,1470(35):98-110.
[3]SHEN Y,QIN H,CHEN J,et al.Postnatal activation of TLR4in astrocytes promotes excitatory synaptogenesis in hippocampal neurons[J].Journal of Cell Biology,2016,215(5):719-734.
[4]孙影,吴兴饶,许志峰,等.粒细胞集落刺激因子对癫痫模型小鼠神经元及神经炎症的影响[J].中国神经精神疾病杂志,2018,44(10):609-612.
[5]MAROSO M,BALOSSO S,RAVIZZA T,et al.Toll-like receptor 4 and high-mobility group box-1 are involved in ictogenesis and can be targeted to reduce seizures[J].Nature Medicine,2010,16(4):413-419.
[6]RODGERS K M,HUTCHINSON M R,NORTHCUTT A,et al.The cortical innate immune response increases local neuronal excitability leading to seizures[J].Brain,2009,132(9):2478-2486.
[7]DAS A,TH W G,HOLMES C,et al.Hippocampal tissue of patients with refractory temporal lobe epilepsy is associated with astrocyte activation,inflammation,and altered expression of channels and receptors[J].Neuroscience,2012,220(38):237-246.
[8]ZHANG Z,LIU Q,MING L,et al.Upregulation of HMGB1-TLR4 inflammatory pathway in focal cortical dysplasia type II[J].Journal of Neuroinflammation,2018,15(1):27.
[9]HU QP,MAO DA.Histone deacetylase inhibitor SAHA attenuates post-seizure hippocampal microglia TLR4/MYD88 signaling and inhibits TLR4 gene expression via histone acetylation[J].Bmc Neuroscience,2016,17(1):22.
[10]WEN X,HAN XR,WANG YJ,et al.MicroRNA-421 suppresses the apoptosis and autophagy of hippocampal neurons in epilepsy mice model by inhibition of the TLR/MYD88 pathway[J].Journal of Cellular Physiology,2018,233(9):7022-7034.
[11]张菲菲,程艳伟,石向群.大鼠癫痫持续状态后海马TNF-α、IL-1β的动态变化[J].中国神经精神疾病杂志,2017,43(6):362-367.