摘要
目的观察加味柴胡疏肝汤对癫痫小鼠海马神经元自噬和凋亡的拮抗作用,探讨其发挥神经保护作用的机制。方法将60只小鼠随机分为6组:生理盐水组(20 mL·kg-1·d-1灌胃两周)、模型组(180mg·kg-1腹腔注射)、加味柴胡疏肝汤组(7 g·kg-1·d-1灌胃两周)、加味柴胡疏肝汤+miRNA-204 mimic组(2μL颅内注射)、加味柴胡疏肝汤+miRNA-204 inhibitor组(2μL颅内注射)、卡马西平组(混悬液30 mg·kg-1·d-1灌胃两周);采用匹罗卡品使小鼠致痫,抑制及过表达miRNA-204后,分别予加味柴胡疏肝汤灌胃,处死小鼠,取其海马组织,观察每组的指标情况:采用实时荧光定量聚合酶链式反应(Real-time PCR)技术检测小鼠海马miRNA-204,以及自噬相关基因ATG7、LC3Ⅱ的表达情况,采用原位缺口末端标记法(TUNEL)检测小鼠海马C1区神经元凋亡情况。结果与生理盐水组相比,模型组小鼠海马miR-204表达明显降低(P<0.01),自噬相关基因ATG7、LC3Ⅱ的表达,神经元凋亡指数明显升高(P<0.01);与模型组相比,使用加味柴胡疏肝汤后,小鼠海马组织miR-204表达明显升高,自噬相关基因ATG7、LC3Ⅱ的表达,神经元凋亡指数明显降低(P<0.05);使miR-204过表达后,与单纯使用柴胡疏肝汤相比,上述指标变化趋势相同,变化幅度增大(P<0.01);抑制miR-204表达后,与单纯使用柴胡疏肝汤相比,上述指标变化趋势相同,变化幅度减少(P<0.05)。结论加味柴胡疏肝汤具有抗癫痫作用,可能是通过升高miRNA-204的表达,拮抗小鼠海马神经元过度自噬和凋亡。
Objective To observe the antagonistic effect of Jiawei Chaihu Shugan Tang(JWCHSGT)on autophagy and apoptosis of hippocampal neurons in epileptic mice,and to explore its mechanism of neuroprotection.Methods Sixty mice were randomly divided into 6 groups:saline group(20 mL·kg-1·d-1 for two weeks),model group(180 mg·kg-1 for intraperitoneal injection),and JWCHSGT group(7 g·kg-1·d-1 for two weeks),JWCHSGT+miRNA-204 mimic group(2μL intracranial injection),JWCHSGT+miRNA-204 inhibitor group(2μL intracranial injection),carbamazepine group(suspension 30 mg·kg-1·d-1 for two weeks);using pilocarpine to cause epilepsy in mice,respectively,add flavor to Bupleurum after intragastric administration,inhibition and overexpression of miRNA-204,the mice were sacrificed and their hippocampus tissues were harvested.The indicators of each group were observed:Real-time Quantitative PCR Detecting System(RT-PCR)was used to detect mouse hippocampal miRNAs-204 expression,and the expression of autophagy-related genes ATG7,LC3Ⅱ,using the in situ nick end labeling(TUNEL)to detect hippocampal neuronal apoptosis in mouse C1 region.Results Compared with the saline group,the expression of miR-204 in the hippocampus of the model group was significantly decreased(P<0.01),and the expression of autophagy-related genes ATG7 and LC3Ⅱwas significantly increased(P<0.01).Compared with the model group,the expression of miR-204 in hippocampus of mice was significantly increased after the application of JWCHSGT,and the expression of autophagyrelated genes ATG7 and LC3Ⅱwas significantly decreased(P<0.05).After overexpression of miR-204,compared with the simple use of JWCHSGT,the above indicators showed the same trend,and the change range increased(P<0.01).After inhibiting the expression of miR-204,and simply using JWCHSGT.In comparison,the above indicators showed the same trend and the magnitude of change decreased(P<0.05).Conclusion JWCHSGT has anti-epileptic effect,which may antagonize excessive autophagy and apoptosis of mouse hippocampal neurons by increasing the expression of miRNA-204.
作者
汪顺贵
玉倩
李华霞
李欢
卢玲
廖现秋
周艳英
宋曦
秦红玲
刁丽梅
Wang Shungui;Yu Qian;Li Huaxia;Li Huan;Lu Ling;Liao Xianqiu;Zhou Yanying;Song Xi;QinHongling;Diao Limei(Graduate School of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China;The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530001,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第7期2154-2159,共6页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会地区项目(81760809):基于miRNA-204通过mTOR通路调控海马神经元内质网自噬探讨加味柴胡疏肝汤对癫痫的疗效机制研究,负责人:刁丽梅
广西壮族自治区科学技术厅面上项目(2017GXNSFAA198294):基于miRNA-204通过mTOR通路调控海马神经元内质网自噬探讨加味柴胡疏肝汤对癫痫的作用机制,负责人:刁丽梅
广西壮族自治区卫生和计划生育委员会广西医疗卫生适宜技术研究与开发项目(s2017049):加味柴胡疏肝汤通过miRNA-204/mTOR通路调控海马细胞自噬治疗颞叶癫痫的临床研究,负责人:刁丽梅
