新型H_2S供体激活PI3K/Akt对癫痫大鼠脑内Bcl-2/Bax蛋白表达的影响  

Effect of New Hydrogen Sulfide on the Expression of Bcl-2/Bax Protein in the Brain of Epileptic Rats by Activating PI3K/Akt Pathway

作　　者：张思远 蔡昕 卢晓桦 铁子慧 叶芷钘 刘丹琼 都昇 雷水生[2] 朱晓琴 Zhang Siyuan;Cai Xin;Lu Xiaohua(Department of Physiology,School of Basic Medical Sciences,Guangzhou Medical Universily,Guangzhou 511436,China)

机构地区：[1]广州医科大学基础医学院生理学教研室,广州511436 [2]广州医科大学附属第五医院皮肤美容科,广州510700

出　　处：《华中科技大学学报（医学版）》2020年第3期296-300,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81870992,No.81870856);广东省自然科学基金资助项目(No.2020A1515010985);广东省科技计划资助项目(No.2017ZC0234,No.2017ZC0261);广州医科大学大学生实验室开放项目(No.2018-9,No.2020-1);广东省大学生创新创业训练计划项目(No.S201910570083);广州医科大学大学生科技创新项目(No.2018A117)。

摘　　要：目的研究新型H_2S供体对癫痫模型大鼠的作用及其可能的机制,即通过激活PI3K/Akt通路、影响Bcl-2/Bax蛋白表达进而减少癫痫大鼠海马神经元凋亡。方法通过腹腔注射戊四氮(pentylenetetrazole,PTZ)的方法建立大鼠癫痫持续状态(status epilepticus,SE)模型,并经侧脑室注射给予不同药物处理,将实验大鼠随机分为5组:正常对照组(Control组)、PTZ致痫组(SE组)、H_2S预处理组(H_2S+SE组)、DMSO预处理组(DMSO+SE组)、抑制剂组(LY294002+SE组)。采用行为学观察记录大鼠癫痫发作情况;植入脑电引流电极记录大鼠海马脑电变化;TUNEL免疫荧光标记法检测大鼠海马组织细胞凋亡情况;Western blot检测大鼠海马组织中PI3K、Akt、Bcl-2、Bax蛋白表达情况。结果行为学观察发现LY294002+SE组大鼠癫痫发作级别最高,DMSO+SE组和SE组次之,H_2S+SE组较低;H_2S+SE组癫痫发作潜伏期最长,DMSO+SE组和SE组次之,而LY294002+SE组最短。脑电图检测提示LY294002+SE组平均波幅最高,其次为DMSO+SE组和SE组,高于H_2S+SE组,对照组无痫样波。TUNEL染色结果显示,与DMSO+SE组和SE组相比,LY294002+SE组海马CA1区凋亡最严重,而H_2S+SE组凋亡细胞较少。Western blot结果显示:PI3K、Akt、Bcl-2/Bax表达量从低到高依次为正常对照组、LY294002+SE组、SE/DMSO+SE组、H_2S+SE组;与SE组相比,H_2S+SE组Bcl-2/Bax表达量显著上调(P<0.05)。结论H_2S通过激活PI3K/Akt通路,改变该通路所介导的凋亡蛋白Bcl-2和Bax的表达比例,从而发挥抑制癫痫、保护海马神经元的作用,这种作用可能与抑制海马神经元的凋亡有关。Objective To study the effect of novel carbazole-based H2S donor on epileptic rats and the possible mechanism.Methods The status epilepticus(SE)rat model was established by intraperitoneal injection of pentylenetetrazole(PTZ).Drugs were injected in the lateral ventricle.The rats were randomly divided into 5 groups:Control group,PTZ epileptic group(SE group),H2S pre-treatment epileptic group(H2S+SE group),Dimethyl sulfoxide(DMSO)pre-treatment epileptic group,inhibitor group(LY294002+SE group).The epileptic seizures of rats were recorded by behavioral observation.Brain electrodes was implanted and electroencephalogram(EEG)was recorded in the hippocampus of rats.TUNEL immunofluorescence assay was used to detect hippocampal apoptosis in rats of each group.Western blotting was used to detect protein expression of PI3 K,Akt,Bcl-2,Bax in hippocampal tissues of rats in each group.Results Behavioral observation showed that the order of epileptic seizure level was as follows:LY294002+SE group>DMSO+SE group>SE group>H2S+SE group,and no seizure was observed in the Control group(P<0.05).Incubation time length of epileptic seizure,ranked in descending order,were as follows:H2S+SE group,DMSO+SE group and SE group,and LY294002+SE group.Average amplitude shown by EEG,ranked in descending order,were as follows:LY294002+SE group,DMSO+SE group and SE group,H2S+SE group,while no epileptic wave was found in the Control group(P<0.05).TUNEL staining results showed that,compared with DMSO+SE group and SE group,the apoptosis in hippocampal CA1 area of LY294002+SE group was the most serious,while the number of apoptotic cells of H2S+SE group was smaller(P<0.05).Western blotting results of protein expression of PI3 K,Akt,Bcl-2/Bax,ranked in an ascending order,were as follows:Control,LY294002+SE,SE/DMSO+SE,H2S+SE group.Compared with SE group,the Bcl-2/Bax expression level was significantly increased in H2S+SE group(P<0.05).Conclusion H2S changes the expression ratio of apoptotic protein Bcl-2 to Bax through activating PI3 K/Akt pathway,there

关 键 词：新型H_2S供体 PI3K/Akt通路 Bcl-2/Bax表达 海马神经元凋亡 癫痫 

分 类 号：R742.1[医药卫生—神经病学与精神病学]