S100B抑制剂减少颞叶内侧癫痫模型鼠海马中的COX-2、IL-1β的表达

S100B Inhibitor Reduces the Expressions of COX-2 and IL-1βin Hippocampus of Rats with Mesial Temporal Lobe Epilepsy

目的探究S100B抑制剂Pentamidine对内侧颞叶癫痫(MTLE)不同时期COX-2、IL-1β蛋白表达的影响及其可能的作用机制。方法160只小鼠随机分为对照组和模型组,模型组小鼠右背海马区给予注射50 nL海人酸-PBS溶液(1μg/μL),对照组小鼠则注射等体积生理盐水。分别于建模前1 d和建模6 h后向各组小鼠尾静脉注射Pentamidine(5mg/kg)或等体积的PBS,每天1次,连续7 d。MTLE成功诱导后12 h(急性期)、7 d(潜伏期)、40 d(慢性期)取小鼠海马组织行后续实验。采用RT-PCR检测小鼠海马组织S100B mRNA表达水平;采用尼氏染色检测小鼠海马组织尼氏小体和神经元丢失;采用Western blot检测小鼠海马组织COX-2和IL-1β蛋白表达。结果与对照-PBS组比较,急性期、潜伏期和慢性期-PBS组海马组织中S100B mRNA表达不同程度增加(P<0.05);与各PBS组比较,不同时期Pentamidine组S100B mRNA的表达下降(P<0.05)。尼氏染色显示,与对照-PBS组比较,急性期、潜伏期和慢性期-PBS组神经元结构不同程度受损,尼氏小体数量下降(P<0.05);与各PBS组比较,不同时期Pentamidine组的神经元受损程度减轻,尼氏小体数量明显增加(P<0.05)。Western blot显示,与对照-PBS组比较,急性期、潜伏期和慢性期-PBS组海马组织COX-2、IL-1β和phospho-NF-κB的蛋白表达增加(P<0.05);与各PBS-组比较,不同时期Pentamidine组COX-2、IL-1β和phospho-NF-κB蛋白表达下调(P<0.05)。结论S100B抑制剂Pentamidine可通过下调COX-2和IL-1β的表达降低MTLE小鼠海马炎症反应和神经损伤,为MTLE的临床治疗提供重要的参考价值。

Objective To investigate the effects of Pentamidine on the expressions of COX-2 and IL-1βin different stages of medial temporal lobe epilepsy(MTLE)and its potential mechanism.Methods 160 mice were randomly divided into the control group and model group.The right hippocampus of the model group was given 50nl of KA-PBS(1μg/μL),while the control group was injected with an equal volume of PBS solution.Each group of mice were injected with Pentamidine(5mg/kg)or an equal volume of PBS once a day for 7 days before modeling and 6 hours after modeling.Subsequent experiments were performed on hippocampus of mice at 12h(acute phase),7d(latency),and 30d(chronic phase)after a successful MTLE induction.The mRNA expression of S100B in mouse hippocampus was detected by RT-PCR.The Nissl bodies and neuronal loss in mouse hippocampus were detected by Nissl staining.The protein expressions of COX-2 and IL-1βwere analyzed by western blot.Results Compared with the control-PBS group,the relative mRNA expression of S100B in hippocampus of the acute phase,latent phase and chronic phase-PBS group was increased with varying degrees(P<0.05).Compared with the PBS-group,the relative expression of S100B mRNA of Pentamidine group in different periods were decreased(P<0.05).Nissl staining showed the neuron structure in the acute phase,latent phase and chronic phase-PBS group was damaged with varying degrees,compared with the control-PBS group,and the number of Nissl bodies decreased.Compared with the PBS group,the neuron damage of Pentamidine group in different periods was reduced and the number of Nissl bodies was increased(P<0.05).Western blot showed that the expressions of COX-2,IL-1βand phospho-NF-κB of the acute phase,latent phase and chronic phase-PBS group were increased significantly,compared with the control-PBS group(P<0.05).Compared with the PBS-group,the expressions of COX-2,IL-1βand phospho-NF-κB of Pentamidine group in different periods were significantly down-regulated(P<0.05).Conclusion Pentamidine can reduce the hippocampal inflammatory response and nerve damage in MTLE mice by down-regulating the expressions of COX-2 and IL-1β,thus providing an important reference value for the clinical treatment of MTLE.