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云南特色中药臭灵丹体外抗甲型H1N1 流感病毒的实验研究
引用本文:夏晓玲,孙强明,王晓丹,赵玉娇,杨子峰,黄庆晖,姜志宏,王新华,张荣平. 云南特色中药臭灵丹体外抗甲型H1N1 流感病毒的实验研究[J]. 中国中药杂志, 2015, 40(18): 3687-3692
作者姓名:夏晓玲  孙强明  王晓丹  赵玉娇  杨子峰  黄庆晖  姜志宏  王新华  张荣平
作者单位:昆明医科大学, 云南 昆明 650500,北京协和医学院 中国医学科学院 医学生物学研究所 云南省重大传染病疫苗研发重点实验室, 云南 昆明 650118,北京协和医学院 中国医学科学院 医学生物学研究所 云南省重大传染病疫苗研发重点实验室, 云南 昆明 650118,北京协和医学院 中国医学科学院 医学生物学研究所 云南省重大传染病疫苗研发重点实验室, 云南 昆明 650118,广州医科大学 附属第一医院 广州医科大学呼吸疾病国家重点实验室, 广东 广州 510230,广州医科大学 附属第一医院 广州医科大学呼吸疾病国家重点实验室, 广东 广州 510230,澳门科技大学 中药质量研究国家重点实验室, 澳门 00853,广州医科大学 附属第一医院 广州医科大学呼吸疾病国家重点实验室, 广东 广州 510230,昆明医科大学, 云南 昆明 650500
基金项目:国家自然科学基金项目(81460593);广州市教育系统创新学术团队项目(13C07);广州市科技计划项目-科技惠民专项(2014Y2-00031);广州市健康医疗协同创新重大专项(201400000002);云南省应用基础研究重点项目(2014FA031)
摘    要:臭灵丹Laggera pterodonta,一种民间常用的治疗感冒的具有民俗特色的中药。为了检测臭灵丹不同溶剂萃取物对甲型H1N1流感病毒在体外的中和杀伤作用和增殖抑制作用,本研究采用中和抑制实验和增殖抑制实验,用狗肾传代MDCK细胞培养法观察臭灵丹萃取物抑制甲型H1N1流感病毒的致细胞病变作用(CPE),观察2种实验中臭灵丹萃取物在体外对甲型H1N1流感病毒血凝效价的影响,同时采用Real time RT-PCR定量检测流感病毒的拷贝数,比较臭灵丹不同溶剂萃取物对甲型H1N1流感病毒在体外增殖的影响。结果显示,臭灵丹乙酸乙酯萃取物及石油醚萃取物作用72 h后,中和实验组H1N1病毒血凝效价下降了8倍,而增殖抑制组,H1N1病毒血凝效价也分别下降了2倍和4倍;Real time RT-PCR的检测结果表明,臭灵丹乙酸乙酯萃取物中和H1N1病毒的抑制率为72.5%,而增殖抑制组的抑制率为25.3%,石油醚萃取物中和H1N1病毒的抑制率为60.2%,增殖抑制组为81.4%。由此可以得出,臭灵丹乙酸乙酯萃取物和石油醚萃取物在体外对甲型H1N1流感病毒有明显的中和作用及直接的抑制增殖作用。

关 键 词:臭灵丹  甲型流感病毒  抗病毒药物  中草药
收稿时间:2015-02-01

Effect of Yunnan herb Laggera pterodonta against influenza A(H1N1)virus in vitro
XIA Xiao-ling,SUN Qiang-ming,WANG Xiao-dan,ZHAO Yu-jiao,YANG Zi-feng,HUANG Qing-hui,JIANG Zhi-hong,WANG Xin-hua and ZHANG Rong-ping. Effect of Yunnan herb Laggera pterodonta against influenza A(H1N1)virus in vitro[J]. China Journal of Chinese Materia Medica, 2015, 40(18): 3687-3692
Authors:XIA Xiao-ling  SUN Qiang-ming  WANG Xiao-dan  ZHAO Yu-jiao  YANG Zi-feng  HUANG Qing-hui  JIANG Zhi-hong  WANG Xin-hua  ZHANG Rong-ping
Affiliation:Kunming Medical University, Kunming 650500, China,Institute of Medical Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Disease, Kunming 650118, China,Institute of Medical Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Disease, Kunming 650118, China,Institute of Medical Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Disease, Kunming 650118, China,The First Affiliated Hospital of Guangzhou Medical University, State Key Laboratory of Respiratory Diseases, Guangzhou 510230, China,The First Affiliated Hospital of Guangzhou Medical University, State Key Laboratory of Respiratory Diseases, Guangzhou 510230, China,Macau University of Science and Technology, State Key Laboratory for Quality Research in Chinese Medicines, Macau 00853, China,The First Affiliated Hospital of Guangzhou Medical University, State Key Laboratory of Respiratory Diseases, Guangzhou 510230, China and Kunming Medical University, Kunming 650500, China
Abstract:Laggera pterodonta is commonly used for treating influenza in Southwest China, especially in Yunnnan province. The main clinical effects of L. pterodonta include anti-influenza, anti-microbial, anti-inflammatory. To investigate the anti-influenza A (H1N1) virus effect of L. pterodonta, neutralization inhibition and proliferation inhibition tests were performed. MDCK culture method was used to observe the cytopathic effect (CPE) of extracts from L. pterodonta in inhibiting influenza A (H1N1) virus and haemagglutination titre of H1N1 virus in vitro. The culture medium were collected at 24 h, 48 h, 72 h, 96 h,and detected by Real time RT-PCR, in order to compare the effect of different extracts from L. pterodonta on in vitro proliferation of H1N1 virus. The result of neutralization inhibition test showed that hemagglutination titer of ethyl acetate extract were 8 times lower at 72 h; in proliferation inhibition test, hemagglutination titer of ethyl acetate extracts reduced by 2 and 4 times. According to the results of Real time RT-PCR test, the H1N1 inhibition ratio of ethyl acetate extract was 72.5%,while the proliferation inhibition ratio of ethyl acetate extract was 25.3%; as for petroleum ether extracts, the H1N1 inhibition ratio was 60.2%, while the proliferation inhibition ratio was 81.4%. In conclusion, both ethyl acetate extract and petroleum ether extract of L. pterodonta have significant neutralization and direct proliferation inhibition effects on influenza A virus.
Keywords:Laggera pterodonta  influenza A virus  antiviral agents  Chinese herbal medicine
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