定量RT-PCR法对“苦参”在心肌炎模型中药效学研究

The Use of Quantitative RT - PCR to Detect the Therapeutic Effect of “Ku -Shen” in Balb/c Myocarditis Model

目的用定量RT-PCR法检测“苦参”中抗柯萨奇B病毒有效成份“抗柯注射液”,在Balb/c小鼠病毒性心肌炎模型病毒血症时,对抗“柯萨奇B病毒”的药效。 方法取48只雄性18-22g 8周龄的Balb/c小鼠,随机分成A-H共8组,每组6只。1.各治疗组(A-F组),每只小鼠从腹腔接种0.1ml l04TCID50 CVB3m病毒2h后,每天从尾静脉分别注射抗柯注射液,5、10、15、20、25、30mg/kg,2次/d,连续用药3d。2.病毒组(G组),注射病毒同上法,2h后从尾静脉注入0.3ml无菌生理盐水,2次/d,连续3d。3.空白对照组(H组),腹腔内注射无小牛血清的RPMI-1640培养液,2h后从尾静脉注入0.3ml无菌生理盐水,2次/d,连续3d。 结果 空白组CVB-RNA(-)对接种CVB3m感染后用不同剂量“抗柯注射液”治疗的各小组血液中CVB3m-RNA含量与病毒组相比明显下降,其抑制率(％)与所用抗柯注射液的剂量呈正比,即“抗柯”剂量增加,CVB3m-RNA含量下降的幅度亦增加,每天5-30mg,/kg中的各剂量都有明显抑制病毒增殖的作用。 结论用定量RT-PCR法检测到“抗柯注射液”对Balb/c小鼠病毒性心肌炎病毒血症时效果明显,最小有效剂量为5mg·kg-1·d-1。

Objective To use quantitative RT-PCR method to study the therapeutic effect of 'Kang - Ke fluid '(an active substance of ' Ku -Shen' on anti - Coxsachie B virus) in virus infected Balb/ c mice at the viremia stage. Methods Forty-eight male,8 weeks( weight 18 - 22g) Balb/c mice were divided into 8 groups (n =6 each) A,B,C ,D,E ,F. For these therapeutic groups, every mouse was inoculated with 0.1ml 10-4 TCID50CVB3m peritoneally. After 2h Kang - Ke injection fluid was injected into the tail vein 5mg · kg-1 · d-1 for group A;10mg ·kg-1 · d-1 for group B;15mg · kg-1 · d-1 for group C;20mg ·kg-1 · d-1 for group D;25mg ·kg-1 · d -1 for group E;30mg ·kg-1 · d-1 for group F, Bid, respectively for 3d. For the viral infected group G; every mouse was inoculated with the same amount of virus as therapeutic group, and after 2h,0. 3ml sterilized normal saline was injected Bid into the tail vein for 3d. For the control group H: every mouse was injected RPMI -1640 0. lml(without calf serum) peritoneally after 2h,0.3ml normal saline was injected Bid into the tail vein for 3d. Results With different doses of 'Kang - Ke injection fluid' , the content of CVB3m RNA decreased significantly. The inhibition rate ( % ) was in proportional to the dose of Kang - Ke, being more and more decreased with increase of the Kang - Ke dosage. Conclusion 'Kang - Ke injection fluid' has significant therapeutic efficacy proved by the quantitative RT -PCR CVB3mRNA method of Balb/c mice myocarditis model at the viremia stage and its smallest effec live dose is 5 mg · kg-1 · d-1