云南特色中药臭灵丹体外抗甲型H1N1流感病毒实验研究.docVIP

云南特色中药臭灵丹体外抗甲型H1N1流感病毒的实验研究 　　[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 　　Effect of Yunnan herb Laggera pterodonta against influenza 　　A（H1N1）virus in vitro 　　XIA Xiao-ling1， SUN Qiang-ming2， WANG Xiao-dan2， ZHAO Yu-jiao2， YANG Zi-feng3， 　　HUANG Qing-hui3， JIANG Zhi-hong4， WANG Xin-hua3*， ZHANG Rong-ping1* 　　（1.Kunming Medical University， Kunming 650500， China; 　　2. Institute of Medical Biology， Chinese Academy of Medical Sciences/Peking Union Medical College， Yunnan Key 　　Laboratory of Vaccine Research amp; Development on Severe Infectious Disease， Kunming 650118， China; 　　3. The First Affiliated Hospital of Guangzhou Medical University， State Key Laboratory of Respiratory Diseases， Guangzhou 510230， China; 　　4.Macau University of Science and Technology， State Key Laboratory for Quality Research in Chinese Medicines， Macau 00853， 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 Re