养猪
養豬
양저
SWINE PRODUCTION
2009年
4期
73-74
,共2页
顾文松%沈明华%刘伟%侯建德
顧文鬆%瀋明華%劉偉%侯建德
고문송%침명화%류위%후건덕
细胞培养%中药复方提取物%猪繁殖与呼吸综合征病毒
細胞培養%中藥複方提取物%豬繁殖與呼吸綜閤徵病毒
세포배양%중약복방제취물%저번식여호흡종합정병독
ceil culture%Chinese herbal extract%PRRSV
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25 mg/mL,稀释成2 500、1 250、625.0、312.5、156.25、78.13、39.06、19.53、9.77、4.88μg/mL.培养细胞选用Marc-145.试验方法:1)细胞培养液中加入0.1 mL中药提取物培养2 h后加入PRRSV病毒液0.1 mL,培养72 h;2)细胞培养液中加入病毒液0.1 mL培养2 h后加入中药提取物0.1 mL,培养72 h;3)同时将各浓度中药提取物0.1 mL和病毒液0.1 mL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,19.53~2 500 μg/mL效果显著,其中39.06 μg/mL效果最好;第2种加药方式下,156.25~2 500 μs/mL效果显著,1 250μg/mL效果最佳;第3种方式下,19.53-2 500 μg/mL效果显著,2 500μg/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.
中藥複方提取物經臨床驗證對預防豬繁殖與呼吸綜閤徵病毒(PRRSV)、豬圓環病毒2型等引起的疾病有效.本試驗測定細胞安全濃度範圍內中藥複方提取物對細胞抗PRRSV感染能力的影響.試驗用中藥主要成分為虎杖、女貞子、槓闆歸等.提取方法為水浸,鏇轉蒸髮榦燥.製成生藥原液濃度為25 mg/mL,稀釋成2 500、1 250、625.0、312.5、156.25、78.13、39.06、19.53、9.77、4.88μg/mL.培養細胞選用Marc-145.試驗方法:1)細胞培養液中加入0.1 mL中藥提取物培養2 h後加入PRRSV病毒液0.1 mL,培養72 h;2)細胞培養液中加入病毒液0.1 mL培養2 h後加入中藥提取物0.1 mL,培養72 h;3)同時將各濃度中藥提取物0.1 mL和病毒液0.1 mL加入細胞培養液中;3種加藥方式均設細胞對照和病毒對照,每箇濃度設4~6箇重複.結果錶明,第1種加藥方式下,19.53~2 500 μg/mL效果顯著,其中39.06 μg/mL效果最好;第2種加藥方式下,156.25~2 500 μs/mL效果顯著,1 250μg/mL效果最佳;第3種方式下,19.53-2 500 μg/mL效果顯著,2 500μg/mL效果最好.這可能是中藥複方提取物預防PRRSV感染的作用機製之一.
중약복방제취물경림상험증대예방저번식여호흡종합정병독(PRRSV)、저원배병독2형등인기적질병유효.본시험측정세포안전농도범위내중약복방제취물대세포항PRRSV감염능력적영향.시험용중약주요성분위호장、녀정자、강판귀등.제취방법위수침,선전증발간조.제성생약원액농도위25 mg/mL,희석성2 500、1 250、625.0、312.5、156.25、78.13、39.06、19.53、9.77、4.88μg/mL.배양세포선용Marc-145.시험방법:1)세포배양액중가입0.1 mL중약제취물배양2 h후가입PRRSV병독액0.1 mL,배양72 h;2)세포배양액중가입병독액0.1 mL배양2 h후가입중약제취물0.1 mL,배양72 h;3)동시장각농도중약제취물0.1 mL화병독액0.1 mL가입세포배양액중;3충가약방식균설세포대조화병독대조,매개농도설4~6개중복.결과표명,제1충가약방식하,19.53~2 500 μg/mL효과현저,기중39.06 μg/mL효과최호;제2충가약방식하,156.25~2 500 μs/mL효과현저,1 250μg/mL효과최가;제3충방식하,19.53-2 500 μg/mL효과현저,2 500μg/mL효과최호.저가능시중약복방제취물예방PRRSV감염적작용궤제지일.
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.