天麻破壁粉、冻干粉对戊四唑诱导癫痫大鼠海马组织Bcl-2、Bax、Caspase-3、GAT-1 mRNA及蛋白表达对比研究  

Comparative Study on the Expression of Bcl-2,Bax,Caspase-3 and GAT-1 mRNA and Protein in Hippocampus of Pentylenetetrazol-induced Epilepsy Rats by Tianma Cell Wall-Broken Decoction Pieces and Freeze-dried Powder

作　　者：刘春艳 柴艺汇 田兴中 唐靖雯 卢礼平 谢宇 周雯 牛建均 陈露露 张丽艳 茅向军[3] Liu Chunyan;Chai Yihui;Tian Xingzhong;Tang Jingwen;Lu Liping;Xie Yu;Zhou Wen;Niu Jianjun;Chen Lulu;Zhang Liyan;Mao Xiangjun(Guizhou College of Traditional Chinese Medicine,Guiyang 550025,China;Guizhou Weimen Pharmaceutical Company Limited,Guiyang 550018,China;Guizhou Provincial Institute of Food and Drug Control,Guiyang 550004,China;The Fifth Hospital of Zhangjiakou City,Zhangjiakou 075000,China;Guizhou Guangjitang Pharmaceutical Company Limited,Guiyang 550014,China)

机构地区：[1]贵州中医药大学,贵阳550025 [2]贵州威门药业股份有限公司,贵阳550018 [3]贵州省食品药品检验所,贵阳550004 [4]张家口市第五医院,张家口075000 [5]贵州广济堂药业有限公司,贵阳550014

出　　处：《世界科学技术-中医药现代化》2020年第1期147-156,共10页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：贵州省科学技术厅黔科合重大专项([2016]3023号):贵州特色、大宗药材破壁技术标准化研究与产业化,负责人:张丽艳;贵州省教育厅国内一流建设学科(GNYL2017]008号):中药学,负责人:周英;贵州省科学技术厅黔科合平台人才([2017]5403):贵州省破壁饮片工程技术研究中心,负责人:谢字。

摘　　要：目的通过研究天麻(普通粉、破壁粉、冻干粉)对戊四唑诱导癫痫大鼠海马组织B细胞白血病-淋巴瘤-2基因(B-cell leukemia-iymphoma-2,Bcl-2),B细胞白血病淋巴瘤相关蛋白基因(Bcl-2-associated X protein gene,Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和γ-氨基丁酸转运体1(GAT-1)基因和蛋白的表达影响对天麻神经保护作用机制进行初步探讨,并对三种加工方式的天麻效应差异性进行对比研究。方法以SD大鼠为研究对象,建立戊四唑癫痫模型。随机分为12组,正常组,戊四唑组,丙戊酸钠组,天麻(普通粉、破壁粉、冻干粉)分别对应的低、中、高剂量组(0.5、1.0、2.0 g·kg-1)连续灌胃10 d。用荧光定量聚合酶链式反应法(Real-time PCR)检测三种加工方式的天麻对戊四唑诱导癫痫海马组织Bcl-2、Bax、Caspase-3和GAT-1基因表达的情况;用蛋白免疫印迹法(Western Blot)检测天麻对戊四唑诱导癫痫海马组织Bax、Bcl-2、Caspase-3和GAT-1蛋白表达情况。结果模型组癫痫发作强度均为Ⅵ级,表明造模成功。与模型组比较,丙戊酸钠组及天麻普通粉、破壁粉、冻干粉各剂量组癫痫潜伏时间显著延长(P<0.05),癫痫持续时间显著缩短(P<0.05)。与丙戊酸钠组比较,天麻普通粉高剂量组、天麻破壁粉高剂量组癫痫潜伏时间无显著差异(P<0.05),而天麻冻干粉高剂量组癫痫潜伏时间显著延长(P<0.05),且持续时间与之无显著性差异(P<0.05)。与正常组比较,模型组Bax、Caspase3和GAT-1mRNA及蛋白相对表达量显著升高(P<0.05),Bcl-2 mRNA及蛋白相对表达量显著下降(P<0.05);与模型组比较,丙戊酸钠组和天麻各组可显著降低Bax、Caspase3和GAT-1 mRNA及蛋白相对表达量(P<0.05),显著升高Bcl-2 mRNA及蛋白相对表达量(P<0.05)。结论天麻冻干粉高剂量组效果最佳,天麻破壁粉高剂量组次之,最后天麻普通粉高剂量组。天麻对神经的保护具有剂量依赖性,并与天麻不同加工方式Objective To study Tianma(general powder,cell wall-broken decoction pieces,Freeze-dried Powder)on rat hippocampus with pentylenetetrazol-induced epilepsy.The expression of B-cell leukemia-iymphoma-2,(Bcl-2),Bcl-2-associated X protein gene(Bax),caspase-3 and gamma-aminobutyric acid transporter 1(GAT-1)and protein affects the mechanisms were analyzed.A comparative study of the differences in the Tianma effect of the processing methods was performed.Methods Pentylenetetrazol convulsion model was established.SD rats were randomly divided into 12 groups,normal group,pentylenetetrazole group,sodium valproate group,Tianma(ordinary powder,cell wall-broken decoction pieces,freeze-dried Powder)corresponding to low,medium and high doses respectively.Groups(0.5,1.0,2.0 g·kg-1)were by gavage 10 days.The expression of Bcl-2,Bax,Caspase-3 and GAT-1 in hippocampus pentylenetetrazol-induced epilepsy of 3 kinds of processing methods of Tianma was detected by real-time PCR.The expression of Bcl-2,Bax,Caspase-3 and GAT-1 in hippocampus pentylenetetrazol-induced epilepsy of 3 kinds of processing methods of Tianma was detected by Western Blot.Results Compared with the normal group,the relative expression of Bax,Caspase-3 and GAT-1 mRNA in the model group was significantly increased(P<0.05),and the relative expression of Bcl-2 mRNA and protein was significantly decreased(P<0.05).Compared with the model group,the sodium valproatec group control The expression of Bax,Caspase-3 and GAT-1 mRNA and protein were significantly decreased in the group and the Tianma group(P<0.05),and the relative expression of Bcl-2 mRNA and protein was significantly increased(P<0.05).Conclusion The high dose group of Tianma Freeze-dried Powder has the best effect,followed by the high dose group of Tianma cell wall-broken decoction pieces,and the high dose group of Tianma ordinary powder again.The protective effect of Tianma on the nerve was dose-dependent and related to different processing methods of Tianma.The mechanism of action was related to the regulati

关 键 词：天麻 癫痫 冻干粉 Bcl-2 CASPASE-3 Bax GAT-1 

分 类 号：R285.5[医药卫生—中药学]