中药复方提取物对细胞抗猪繁殖与呼吸综合征病毒感染能力的影响

中药复方提取物经临床验证对预防猪繁殖与呼吸综合征病毒（PRRSV）、猪圆环病毒2型等引起的疾病有效。本试验测定细胞安全浓度范围内中药复方提取物对细胞抗PRRSV感染能力的影响。试验用中药主要成分为虎杖、女贞子、杠板归等。提取方法为水浸，旋转蒸发干燥。制成生药原液浓度为25mg／mL，稀释成2500、1250、625．0、312．5、156．25、78．13、39．06、19．53、9．77、4．88ug／mL。培养细胞选用Marc-145。试验方法：1）细胞培养液中加入0．1mL中药提取物培养2h后加入PRRSV病毒液0．1mL，培养72h；2）细胞培养液中加入病毒液0．1mL培养2h后加入中药提取物0．1mL，培养72h；3）同时将各浓度中药提取物0．1mL和病毒液0．1mL加入细胞培养液中；3种加药方式均设细胞对照和病毒对照，每个浓度设4～6个重复。结果表明，第1种加药方式下，19．53—2500ug／mL效果显著，其中39．06ug／mL效果最好；第2种加药方式下，156．25～2500ug／mL效果显著，1250ug／mL效果最佳；第3种方式下，19．53—2500ug／mL效果显著，2500ug／mL效果最好。这可能是中药复方提取物预防PRRSV感染的作用机制之一。

Anti-infectious Potential of Mare-145 Cell Treated by Chinese Herbal Extract in PRRSV Challenge Test

Chinese herbal extract has been confirmed to be effective on precautions of diseases induced by porcine reproductive and respiratory syndrome virus (PRRSV),porcine circus virus (PCV-2),etc.The target of this test was to determine the effect of Chinese herbal extract on anti-infectious potential of Marc-145 cell in PRRSV challenge,within the consistency scope of cell safety.Polygonum Cuspidatum Sieb.et Zucc,Nv-zhenzi,Herba Polygoni Perfoliati and other 7 kinds of Chinese herbs were chosen for the aqueous extraction.Then the stock solution were concentrated by rotary evaporation to 25 mg/mL and diluted to 10 levels:2 500,1 250,625.0,312.5,156.25,78.13,39.06,19.53,9.77,4.88 μg/mL.The injection method followed by: 1)injecting 0.1 mL dose into the Marc-145 cell culture and hatching 2 h,then injecting 0.1 mL PRRSV(100 TCID50),4 replicates for each level;2)injecting 0.1 mL PRRSV and hatching 2 h,then injecting 0.1 mL dose, 4 replicates for each level;3)injecting 0.1 mL dose and 0.1 mL PRRSV at the same time,6 replicates for each level, and there were both virus and cell control with each treatment. Results showed that 19.53-2 500 μg/mL dose may work significantly and 39.06 μg/mL do the best with method 1;156.25-2 500 μg/mL dose may work remarkably and 1 250 μg/mL do the best with method 2;with method 3,19.53～2 500 μg/mL may work efficiently and 2 500 μg/mL do the best.This could be one mechanism of action of Chinese herbal extract to prevent PRRSV infection.