毛蕊异黄酮通过Cyt-C/Apaf-1信号通路抑制实验性自身免疫性脑脊髓炎小鼠中枢神经细胞凋亡  

Calycosin inhibits central nerve cell apoptosis in mice with experimental autoimmune encephalomyelitis through Cyt-C/Apaf-1 signaling pathway

作　　者：杨姜德 刘勇华 朱玉华 陈绪龙 许静 李静 周盈 杨少林 蒋超 YANG Jiangde;LIU Yonghua;ZHU Yuhua;CHEN Xulong;XU Jing;LI Jing;ZHOU Ying;YANG Shaolin;JIANG Chao(Department of Pharmacy,Affiliated Hospital of Jiujiang University,Jiujiang 332000,China)

机构地区：[1]九江学院附属医院药学部,九江332000

出　　处：《中国免疫学杂志》2023年第8期1580-1584,共5页Chinese Journal of Immunology

基　　金：江西省卫生计生委科技计划项目(20181943)。

摘　　要：目的:观察毛蕊异黄酮(CA)对实验性自身免疫性脑脊髓炎(EAE)小鼠中枢神经细胞凋亡的影响,并探索其可能的作用机制。方法:取45只C57BL/6小鼠,随机选取10只作为假手术组,剩余小鼠建立EAE模型,建模成功后随机分为EAE组(10只)、CA组(10只)、联合组(11只)。联合组腹腔注射CA(20 mg/kg),尾静脉注射细胞色素C(Cyt-C)激动剂抗霉素A(AA,100μg/kg);CA组腹腔注射CA(20 mg/kg),尾静脉注射等体积PBS;假手术组、EAE组腹腔注射等体积PBS,尾静脉注射等体积PBS,1次/d,持续14 d。进行神经障碍评分;检测脊髓组织三磷酸腺苷(ATP)含量;透射电镜观察线粒体超微结构;Tunel染色观察脊髓神经细胞凋亡;HE染色观察脊髓病理改变;Western blot检测脊髓组织Cyt-C、凋亡蛋白酶活化因子(Apaf-1)蛋白表达。结果:与EAE组比较,CA组小鼠神经障碍评分、脊髓神经细胞凋亡率、Cyt-C、Apaf-1蛋白表达降低,脊髓组织ATP水平升高(P<0.05);与CA组比较,联合组小鼠神经障碍评分、脊髓神经细胞凋亡率、Cyt-C、Apaf-1蛋白表达升高,脊髓组织ATP水平降低(P<0.05)。结论:CA可减少EAE小鼠中枢神经细胞凋亡,促进脊髓损伤修复,减轻线粒体损伤,可能通过抑制Cyt-C/Apaf-1信号通路发挥作用。Objective:To observe effect of calycosin(CA)on central nerve cell apoptosis in mice with experimental autoimmune encephalomyelitis(EAE),and to explore its possible mechanisms.Methods:Ten mice were randomly selected from 45 C57BL/6 mice as sham operation group,and remaining mice were established EAE model,which were randomly divided into EAE group(10 mice),CA group(10 mice),combined group(11 mice)after successful modeling.Mice in combined group were injected with CA(20 mg/kg)intraperitoneally,and injected with cytochrome C(Cyt-C)agonist antimycin A(AA,100μg/kg)in tail vein.In CA group,CA(20 mg/kg)was injected intraperitoneally,and an equal volume of PBS was injected into tail vein.Sham operation group and EAE group were intraperitoneally injected with an equal volume of PBS,and tail vein was injected with an equal volume of PBS.Once a day for 14 days.Neurological disorder score was assessed.Content of adenosine triphosphate(ATP)in spinal cord tissue was tested.Ultrastructure of mitochondria was observed by transmission electron microscopy.Tunel staining was used to observe apoptosis of spinal cord nerve cells.Pathological changes of spinal cord were observed by HE staining.Western blot was used to detect expressions of Cyt-C and Apaf-1 proteins in spinal cord tissues.Results:Compared with EAE group,mice neurological disorder score,spinal cord neuronal apoptosis rate,Cyt-C,Apaf-1 protein expressions were decreased,and ATP level of spinal cord tissues was increased in CA group(P<0.05).Compared with CA group,mice neurological disorder score,spinal cord neuronal apoptosis rate,Cyt-C,Apaf-1 protein expressions were increased,and ATP level of spinal cord tissue was decreased in combined group(P<0.05).Conclusion:CA can reduce apoptosis of central nerve cells in EAE mice,promote repair of spinal cord injury and reduce mitochondrial damage,which may be achieved by inhibiting Cyt-C/Apaf-1 signaling pathway.

关 键 词：毛蕊异黄酮 细胞色素C 凋亡蛋白酶活化因子 实验性自身免疫性脑脊髓炎 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]