木犀草素对癫痫持续状态幼鼠海马组织神经细胞保护作用的机制研究  

Protective effect of luteolin on hippocampal neurons in juvenile rats with status epilepticus and its mechanism

作　　者：周琴[1] 孔海波 王艳茹 何保梅 ZHOU Qin;KONG Haibo;WANG Yanru;HE Baomei(Department of Pediatrics,Zhejiang Provincial People's Hospital(People's Hospital Affiliated to Hangzhou Medical College),Hangzhou 310014,China)

机构地区：[1]浙江省人民医院(杭州医学院附属人民医院)儿科,杭州310014

出　　处：《浙江医学》2022年第14期1481-1485,1490,I0004,I0005,共8页Zhejiang Medical Journal

基　　金：浙江省实验动物科技计划项目(2018C37086);浙江省中医药科学研究基金计划项目(2020ZB022);浙江省医药卫生科技计划项目(2022KY073)。

摘　　要：目的探讨木犀草素对癫痫持续状态(SE)幼年大鼠(下称幼鼠)海马组织神经细胞保护作用的机制。方法将156只21日龄雄性SD幼鼠按随机数字表法分为对照组12只、模型组48只、木犀草素1组(低剂量20 mg/kg)48只和木犀草素2组(高剂量50 mg/kg)48只,其中模型组、木犀草素1组和木犀草素2组在癫痫发作后的1、2、3和7 d分别处死12只幼鼠。采用氯化锂-匹罗卡品法制作幼鼠SE模型,木犀草素1组和木犀草素2组在造模成功后,腹腔注射相应剂量的木犀草素,此后每天重复注射1次,直至处死。采用qRT-PCR法检测幼鼠海马组织中NF-κB、Toll样受体4(TLR4)、半胱氨酸门冬氨酸特异性蛋白酶3(Caspase-3)、TNF-αmRNA表达水平,Western blot法检测幼鼠海马组织中TLR4蛋白表达水平,TUNEL法观察幼鼠海马CA1区神经细胞凋亡情况。结果与对照组比较,癫痫发作后1、2、3和7 d模型组幼鼠海马组织中TLR4、NF-κB、Caspase-3、TNF-αmRNA及TLR4蛋白表达水平均升高(均P<0.01);与模型组比较,癫痫发作后1、2、3和7 d木犀草素2组幼鼠海马组织中TLR4、NF-κB、Caspase-3、TNF-αmRNA及TLR4蛋白表达水平均降低(均P<0.05),癫痫发作后3和7 d木犀草素1组幼鼠海马组织中TLR4 mRNA表达水平均降低(均P<0.05),癫痫发作后1和3 d木犀草素1组幼鼠海马组织中Caspase-3 mRNA表达水平均降低(均P<0.05),癫痫发作后3 d木犀草素1组幼鼠海马组织中TNF-αmRNA表达水平降低(P<0.01),癫痫发作后7 d木犀草素1组幼鼠海马组织中TLR4蛋白表达水平降低(P<0.05)。与对照组比较,癫痫发作后1、2、3和7 d模型组大鼠海马组织中神经细胞凋亡率均升高(均P<0.01);与模型组比较,癫痫发作后1、2、3和7 d木犀草素2组幼鼠海马组织神经细胞凋亡率均降低(均P<0.01),癫痫发作后2、3和7 d木犀草素1组幼鼠海马组织神经细胞凋亡率均降低(均P<0.05)。结论木犀草素可减轻SE幼鼠海马组织的炎症反应和细胞凋亡Objective To investigate the protective effect of luteolin on hippocampal neurons in juvenile rats with status epilepticus(SE)and its association with TLR4/NF-κB signaling pathway.Methods One hundred and fifty-six 21-day-old male SD rats were randomly divided into control group(n=12),model group(n=48),low luteolin group(n=48)and high luteolin group(n=48).The SE was induced by the lithium chloride-pilocarpine method.After the successful modeling,20 mg/kg or 50 mg/kg luteolin were injected intraperitoneally(q.d for 2 d)in low and high luteolin groups,respectively.A batch of 12 rats in model group,low and high luteolin groups were sacrificed at 1,2,3 and 7 days after epileptic seizure,respectively.qRT-PCR was used to detect mRNA expression of NF-κB,Toll-like receptor 4(TLR4),TNF-α,cysteinecontaining aspartate-specific proteases-3(Caspase-3);Western blot was used to detect TLR4 protein expression in hippocampus;TUNEL was used to detect hippocampal CA1 neuronal apoptosis.Results Compared with the control group,the expression levels of TLR4,NF-κB,Caspase-3,TNF-α mRNA and TLR4 protein in the hippocampus of the model group were increased at 1,2,3 and 7 days after epileptic seizure(all P<0.01).Compared with the model group,the expression levels of TLR4,NF-κB,Caspase-3,TNF-α mRNA and TLR4 protein in the hippocampus of high luteolin group were decreased at 1,2,3 and 7 days after epileptic seizure(all P<0.05);the expression levels of TLR4 mRNA were in the hippocampus of low luteolin group were decreased at 3 and 7 days after the seizure(all P<0.05),and the Caspase-3 mRNA expression levels in the hippocampus of low luteolin group were decreased at 1,3 days after epileptic seizure(all P<0.05),TNF-α mRNA expression levels in the hippocampus of low luteolin group were decreased 3 days after epilepsy(P<0.01),and the expression level of TLR4 protein in the hippocampus of low luteolin group was decreased 7 days after epileptic seizure(P<0.05).Compared with the control group,the apoptosis rate of neurons in the hippocampus o

关 键 词：木犀草素 癫痫持续状态 凋亡 炎症反应 TOLL样受体4 

分 类 号：R285[医药卫生—中药学]