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戊四唑癫痫模型小鼠急性期海马神经炎症和神经元损伤

Hippocampal neuroinflammation and neuronal injury in the acute phase of pentylenetetrazol induced epilepsy mouse model

  • 摘要:
    目的 探讨戊四唑(pentylenetetrazol,PTZ)诱导癫痫模型小鼠急性痫样发作后24 h海马小胶质细胞和星形胶质细胞活化、促炎因子分泌以及神经元存活情况。
    方法 采用随机数字法将成年雄性C57BL/6小鼠随机分为对照组和PTZ组,每组28只。PTZ组诱导小鼠急性痫样发作,并于发作后24 h使用免疫荧光评估海马CA1区小胶质细胞和星形胶质细胞的活化状态,提取海马组织RNA检测炎症因子mRNA表达水平,采用HE染色评估海马组织神经元的存活情况。
    结果 PTZ组CA1区Iba1+小胶质细胞(55.72±4.29)个 vs (35.71±9.66)个,P<0.001和GFAP+星形胶质细胞(51.61±8.21)个 vs (37.64±5.27)个,P<0.01活化数量明显多于对照组;小胶质细胞活化表型M1型占比显著高于对照组(0.58±0.02 vs 0.35±0.08,P0.0001),M2型占比明显低于对照组(0.25±0.08 vs 0.44±0.09,P<0.01)。PTZ组海马组织中促炎因子IL-1β mRNA相对水平显著高于对照组(2.49±1.61 vs 1.02±0.68,P<0.05);急性期CA1区(284.91±73.32)个 vs (498.88±176.79)个,P<0.05及CA3区(247.42±50.20)个 vs (457.78±159.08)个,P<0.05存活神经元少于对照组。
    结论 PTZ诱导癫痫模型小鼠急性痫样发作后24 h,海马CA1区小胶质细胞和星形胶质细胞激活,小胶质细胞以活化表型M1型为主发挥促炎作用,促炎因子分泌增加,存活的海马神经元数量减少。

     

    Abstract:
    Objective To investigate the activation of microglia and astrocytes, the secretion of pro-inflammatory factors, and the survival of neurons in the hippocampus of mice with acute seizures induced by pentylenetetrazol (PTZ) 24 hours after the onset of seizures.
    Methods Adult male C57BL/6 mice were randomly assigned to the control group and the PTZ-induced acute epileptic seizure group using random numbers, with 28 mice in each group. The activation status of microglia and astrocytes in the CA1 region of the hippocampus was evaluated by immunofluorescence 24 hours after the onset of seizures. RNA was extracted from the hippocampal tissue to detect the expression level of inflammatory factor mRNA, and HE staining was used to assess the survival of neurons in the hippocampus.
    Results Twenty-four hours after PTZ-induced acute seizures in mice, the numbers of activated Iba1+ microglia (55.72±4.29 vs 35.71±9.66, P<0.001) and GFAP+ astrocytes (51.61±8.21 vs 37.64±5.27, P<0.01) in the CA1 region were significantly increased compared with the control group; the proportion of M1 microglia was significantly increased (0.58±0.02 vs 0.35±0.08, P0.0001), while the proportion of M2 microglia was significantly decreased (0.25±0.08 vs 0.44±0.09, P<0.01); the mRNA level of pro-inflammatory factor IL-1β in the hippocampus was significantly higher than that in the control group (2.49±1.61 vs 1.02±0.68, P<0.05); the numbers of neurons in the CA1 (284.91±73.32 vs 498.88±176.79, P<0.05) and CA3 (247.42±50.20 vs 457.78±150.08, P<0.05) regions during the acute phase was significantly reduced compared with the control group.
    Conclusions Twenty-four hours after PTZ-induced acute epileptic seizures in mice, microglia and astrocytes in the CA1 region of the hippocampus are activated, with microglia mainly presenting the M1 phenotype to exert pro-inflammatory effects, secretions of pro-inflammatory factors increasing, and surviving hippocampal neurons decreasing.

     

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