摘要
目的:以热水浴结合脂多糖(LPS)诱导的SD幼鼠高热惊厥(Febrile seizure, FS)模型探究羚羊角抗高热惊厥的作用。方法:72只21 d龄雄性SD大鼠,随机分为正常对照组、模型对照组、羚羊角1、1.5、3 g/kg组和地西泮6.7 mg/kg组。各组大鼠预先灌胃相应药物3 d,末次灌胃前1 h腹腔注射LPS。灌胃后1.5 h,造模幼鼠用热水浴(46±0.5)℃诱导惊厥。观察并记录惊厥延迟时间、惊厥评分、惊厥死亡率;尼氏染色观察大鼠脑组织病变情况;ELISA法检测大鼠脑组织中神经递质谷氨酸(Glu)、去甲肾上腺素(NE)、γ-氨基丁酸(GABA)含量、白介素1β(IL-1β)、IL-6、肿瘤坏死因子(TNF)-α、IL-18含量及大鼠海马中半胱氨酸天冬氨酸蛋白酶1(Caspase-1)蛋白含量;RT-PCR法检测大鼠海马组织中Caspase-1、核苷酸结合寡聚化结构域样受体蛋白3(Nlrp3)、凋亡相关微粒蛋白(Asc)、Il1b、Il18 mRNA表达;Western blot法检测海马组织中细胞核与细胞质中核转录因子-κB(NF-κB)蛋白表达。结果:与正常对照组相比,模型对照组惊厥评级、死亡率明显增加,惊厥延迟时间明显降低(P<0.05或P<0.01),脑组织中Glu、IL-1β、IL-18、IL-6和TNF-α含量明显增加,NE和GABA含量明显降低(P<0.05或P<0.01),脑组织中Caspase-1蛋白含量显著增加,海马中Caspase-1、Nlrp3、Asc、Il1β及Il18 mRNA表达显著上调,海马组织核NF-κB表达显著上调(P<0.01),胞NF-κB表达下调(P<0.01);与模型对照组相比,羚羊角3 g/kg组能显著延长FS大鼠惊厥延迟时间,降低惊厥评分和惊厥死亡率(P<0.01),且对高热惊厥造成的神经病理损伤均有明显改善;羚羊角各组脑组织中Glu、GABA含量显著降低,NE含量显著升高(P<0.01);IL-1β、IL-6、TNF-α和IL-18含量明显降低(P<0.05),海马Caspase-1、Nlrp3、Asc、Il1b及Il18 mRNA表达均明显下调(P<0.01或P<0.05)。羚羊角3 g/kg组明显下调FS幼鼠海马组织中NF-κB蛋白的核转位并降低脑中Caspase-1蛋白含量(P<0.01)。结�
Objective:To investigate the effect of Lingyangjiao(羚羊角)on febrile seizures(FS)in young SD rats induced by warm water bath combined with lipopolysaccharide(LPS).Methods:Seventy-two male SD rats of 21 days old were randomly assigned into a normal control group,a model group,Lingyangjiao(1,1.5,and 3 g/kg)groups,and a diazepam(6.7 mg/kg)group.Rats in each group were administrated with corresponding drugs for 3 days,and LPS was intraperitoneally injected 1 h before the last time of administration.FS was induced by warm water bath at(46±0.5)℃1.5 h after administration.The FS delay time,score,and mortality were recorded.Nissl’s staining was employed to reveal the pathological changes of rat brain tissue.Enzyme-linked immunosorbent assay was employed to measure the levels of glutamate(Glu),norepinephrine(NE),γ-aminobutyric acid(GABA),interleukin-1β(IL-1β),IL-6,tumor necrosis factor(TNF)-α,and IL-18 in rat brain tissue and the level of cysteinyl aspartate-specific proteinase 1(Caspase-1)in the hippocampus.RT-PCR was employed to determine the mRNA levels of Caspase-1,nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3(Nlrp3),apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain(Asc),Il-1β,and Il-18 in the hippocampus,and western blotting to determine the protein levels of nuclear transcription factor-κB(NF-κB)in the nucleus and cytoplasm of hippocampus.Results:Compared with the normal control group,the model group showed increased FS score and mortality,shortened FS delay time(P<0.05 or P<0.01),elevated levels of Glu,IL-1β,IL-18,IL-6,and TNF-α,and lowered levels of NE and GABA in the brain tissue(P<0.05 or P<0.01).Furthermore,the modeling up-regulated the protein level of Caspase-1 in the brain tissue,the mRNA levels of Caspase-1,Nlrp3,Asc,Il-1β,and Il-18 in the hippocampus tissue,and the protein level of NF-κB in the nucleus of the hippocampus(P<0.01),while it down-regulated the protein level of NF-κB in the cytoplasm
作者
孙雪
张礼林
王成维
李金虎
叶洵
宋林江
吴纯洁
SUN Xue;ZHANG Lilin;WANG Chengwei;LI Jinhu;YE Xun;SONG Linjiang;WU Chunjie(Chengdu University of Traditional Chinese Medicine,Chengdu 615000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2023年第5期82-88,共7页
Pharmacology and Clinics of Chinese Materia Medica
基金
四川省科技厅项目(编号:2021ZYCD012)
成都中医药大学杏林学者苗圃人才专项(编号:MPRC2021012)
成都中医药大学科技园创新创业项目(编号:KJYYB2212)。
关键词
羚羊角
高热惊厥
神经递质
神经炎症
细胞焦亡
Lingyangjiao(羚羊角)
Febrile seizure
Neurotransmitter
Neuroinflammation
Pyroptosis
