硫化氢对局灶性脑缺血损伤大鼠脑组织神经元凋亡的影响  

Effects of hydrogen sulfide on neuron apoptosis of in brain tissue in rats with focal cerebral ischemia injury

作　　者：李国风[1] 张勤增[1] 解丽君[1] 姜红[1] 李立萍[1] 郝娜[1] 张建新[1] LI Guofeng;ZHANG Qinzeng;XIE Lijun(Institute of Pharmaceutical Reserch,Hebei Center for Disease Control and Prevention,Hebei,Shijiazhuang 050021,China)

机构地区：[1]河北省疾病预防控制中心药物研究所,石家庄市050021

出　　处：《河北医药》2020年第8期1135-1139,共5页Hebei Medical Journal

基　　金：河北省自然科学基金(编号:H2014303005);河北省科技厅应用基础研究平台项目(编号:179676127D)。

摘　　要：目的观察硫化氢(H2S)对大鼠局灶性脑缺血损伤所致的神经元凋亡的影响,从神经元凋亡途径探讨H2S对脑缺血损伤的作用及其机制。方法健康雄性SD大鼠共40只,随机分为假手术组、缺血模型组、H2S低剂量组、H2S中剂量组、H2S高剂量组,每组8只。10%的水合氯醛350 mg/kg麻醉大鼠,采用线栓法复制大鼠大脑中动脉闭塞模型。H2S低、中、高剂量组分别于大鼠脑缺血3 h时腹腔注射0.7、1.4、2.8 mg/kg的硫氢化钠(NaHS),假手术组和缺血模型组注射等容量的0.9%氯化钠溶液。5组大鼠均于缺血24 h腹主动脉取血并断头取脑,HE染色法观察脑组织形态学变化,Tunel法检测神经细胞凋亡率,免疫组织化学法检测Bcl-2和Bax在脑组织中的定位和表达,Western blot法分析Caspase-3在脑组织中表达的变化。结果与缺血模型组比较,H2S中、高剂量组大鼠神经细胞受损程度明显减轻,神经细胞凋亡率明显降低(P<0.01),脑组织Bax和Caspase-3蛋白表达明显下降(P<0.01),Bcl-2阳性细胞数明显增多(P<0.01)。结论H2S可以明显降低局灶性脑缺血损伤引起的神经元凋亡,其抗凋亡作用可能与降低Bax和Caspase-3蛋白表达、增加Bcl-2蛋白表达有关。Objective To investigate the effects of hydrogen sulfide(H2S)on neuron apoptosis of in brain tissue in rats with focal cerebral ischemia injury,and to explore its action mechanism.Methods Forty male SD rats(weighed 250~280g)were randomly divided into five groups:sham operation group,ischemia model group,H2S low,middle and high dose groups,with 8 rats in each group.All the rats were anesthetized by 10%chloral hydrate at 350mg/kg,and the animal models with focal cerebral ischemia were established by inserting cranially a nylon thread with rounded tip into internal carotid artery.The rats in sham operation group underwent surgical operation without modeling.The rats in H2S low,middle and high dose groups were respectively administrated with H2S at the dose of 0.7mg,1.4mg and 2.8mg/kg at 3 hours after modeling.However the rats in sham operation group and ischemia model group were given equal volume of 0.9%sodium chloride solution.These rats were sacrificed at 24h after ischemia.The pathological changes of brain tissue were observed under light microscope after HE staining.The neuronal apoptosis was detected by TUNEL assay.The expression levels of Bcl-2 and Bax in brain tissue were detected by immunohistochemisty.Moreover the expression levels of Caspase-3 in the brain tissue were detected by Western Blot.Results As compared with those in ischemia model group,the pathological changes in H2S middle and high dose groups were obviously relieved,and the apoptotic rate of neurons was significantly decreased(P<0.01).The expression levels of Bax and Caspase-3 were significantly decreased in H2S middle and high dose groups,however,the positive cell counts of Bcl-2 were significantly increased(P<0.01).Conclusion The administration of H2S can decrease obviously the apoptotic rate of neurons in brain tissue of rats with focal cerebral ischemic injury,and its action mechanism may be correlated with the decrease of the expression levels of Bax and Caspase-3,and the increase of the expression levels of Bcl-2.

关 键 词：硫化氢 局灶性脑缺血 细胞凋亡 大鼠 

分 类 号：R743.31[医药卫生—神经病学与精神病学]