丁苯酞对慢性低O_(2)高CO_(2)大鼠脑内线粒体凋亡和自噬的影响  

Effects of butylphthalide on mitochondrial apoptosis and autophagy in brain on chronic low O_(2) and high CO_(2) rats

作　　者：闵晶晶 陈琪[2] 顾群[1] 王小同[3] MIN Jingjing;CHEN Qi;GU Qun;WANG Xiaotong(Department of Neurology,the First People′s Hospital of Huzhou,Huzhou,Zhejiang 313000,China;Department of Nephrology,the First People′s Hospital of Huzhou,Huzhou,Zhejiang 313000,China;Department of Rehabilitation,the Second Affiliated Hospital of Wenzhou Medical University,Wenzhou,Zhejiang 325027,China)

机构地区：[1]浙江省湖州市第一人民医院神经内科,313000 [2]浙江省湖州市第一人民医院肾内科,313000 [3]温州医科大学附属第二医院康复科,浙江温州325000

出　　处：《重庆医学》2022年第11期1835-1840,共6页Chongqing medicine

基　　金：浙江省医药卫生科技项目(2020RC118,2021KY1097);浙江省湖州市科技局项目(2019GZ40)。

摘　　要：目的观察丁苯酞胶囊对慢性低氧高二氧化碳(低O_(2)高CO_(2))学习记忆障碍大鼠的影响和潜在机制。方法将64只无特定病原体(SFP)级SD大鼠分为对照组、模型组、植物油组和丁苯酞组,每组16只。除对照组外,模型组、植物油组、丁苯酞组均置于低O_(2)高CO_(2)密闭舱中暴露2周,每天8 h,2周后采用Morris水迷宫、透射电镜、原位末端转移酶标记技术(TUNEL)试剂盒、Western blot检测相关指标。结果在Morris水迷宫实验中,模型组、植物油组大鼠潜伏期都较对照组延长(P<0.05),丁苯酞组与模型组相比潜伏期缩短(P<0.05);各组大鼠穿越平台次数比较差异无统计学意义(P>0.05)。电镜下,低O_(2)高CO_(2)暴露导致线粒体结构破坏,线粒体空泡化,嵴断裂或溶解,细胞内自噬体增加。给予丁苯酞干预后上述情况改善,但细胞内自噬体进一步增加。TUNEL检测法检测显示模型组、植物油组凋亡细胞较对照组增加(P<0.05),丁苯酞组较模型组减少(P<0.05)。Western blot显示,低氧可以诱导自噬相关蛋白及凋亡相关蛋白的表达,给予丁苯酞干预后自噬相关蛋白低氧诱导因子-1α亚基(HIF-1α)、腺病毒E1B 19-kD相互作用蛋白3(BNIP3)、自噬相关蛋白酵母ATG6同源物(Beclin-1)、自噬标记蛋白-微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)及抗凋亡蛋白B淋巴细胞瘤-2基因(Bcl-2)表达进一步增加(P<0.05),促凋亡的Bcl2关联X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(caspase-3)表达减少(P<0.05)。结论丁苯酞可以改善慢性低O_(2)高CO_(2)导致的大鼠空间学习记忆障碍,这可能与其提高线粒体自噬同时降低凋亡有关。Objective To observe the effect and potential mechanism of butylphthalide capsule on the rats with chronic low O_(2) and high CO_(2) learning and memory disorders.Methods Sixty-four SFP grade SD rats were divided into the control group,model group,vegetable oil group and butylphthalide group,16 cases in each group.Except the control group,the model group,vegetable oil group and butylphthalide group were placed in the low O_(2) and high CO_(2) airtight chamber and exposed for 2 weeks,8 h every day.After two weeks,the Morris water maze,transmission electron microscope,TUNEL kit,and Western blot were applied for detecting the related indicators.Results In the Morris water maze,the incubation period in the model group and vegetable oil group was prolonged compared with the control group(P<0.05),but which in the butylphthalide group was shortened compared with the model group(P<0.05).There was no statistical difference in the times of crossing platforms among the various groups(P>0.05).Under the electron microscopy,the low O_(2) and high CO_(2) exposure resulted in the destruction of mitochondrial structure,mitochondrial vacuolization,cristae fracture or dissolution and intracellular autophagosomes increase.The above situation was improved after giving the butylphthalide intervention,but the intracellular autophagosomes further increased.The TUNEL assay detection showed that the apoptosis cells in the model group and vegetable oil group were increased compared with the control group(P<0.05),but which in the butylphthalide group was decreased compared with the model group(P<0.05).Western blot showed that hypoxia could induce the expression of autophagy-associated proteins and apoptosis-related proteins.After giving the butylphthalide intervention,the expressions of autophagy-related proteins HIF-1α,BNIP3,Beclin-1,LC3Ⅱand the anti-apoptosis protein Bcl-2 were further increased(P<0.05).The pro-apoptotic protein Bax and caspase-3 expressions were decreased(P<0.05).Conclusion Butylphthalide may improve the spatial learn

关 键 词：丁苯酞 低氧高二氧化碳氧 凋亡 线粒体自噬 大鼠 

分 类 号：R741.05[医药卫生—神经病学与精神病学]