药物抗CBV3及ECHO11的体外实验研究

目的 寻找抗肠道病毒（EV）感染的安全有效的药物。方法 采用细胞形态观察、MTT法经色检测利巴韦林、双黄连、大蒜素的细胞毒性，并以观察CPE、MTT法比色及蚀斑抑制实验判断其抗CBV3和ECHO11的活性和进行三者间及后二者病毒吸附前后的药效比较。结果 ①利巴韦林TC50为2mg/ml,在1mg/ml-1.5mg/ml时有抑制CBV3和ECHO11的活性。②双黄连TC50为5mg/ml，在0.5mg/ml时即有抑制CBV3和ECHO11的活性，且与药物浓度呈正相关。③大蒜素TC50为12.5μg/ml，在2.5μg/ml-7.5μg/ml时有抑制CBV3和ECHO11的活性。④1.5mg/ml利巴韦林对CBV3及ECHO11的蚀斑抑制率分别为43.2%和37.2%,2.5mg/ml双黄连为81.1%和88.0%,5μg/ml大蒜素为66.2%和77.4%。⑤双黄连在病毒吸附前用药蚀斑抑制率高于吸附手用药（P＜0.05）；大蒜素的差异则不显著（P＞0.05）。结论 三种药物均有体外抗CBV3、ECHO11的作用，但双黄连、大蒜素优于利巴韦林。三种药物中以双黄连毒性最小，抑制病毒活性最高。双黄连在病毒吸附前用药抗病毒活性优于病毒吸附后，有一定预防作用。

The experimental study of the anti-enterovirus effects of drugs %in vitro

OBJECTIVE: To screen the safe and effective anti-enterovirus drugs for clinical application. METHODS: The cytotoxicity of Ribavirin, Shuanghuanglian and Garlic were evaluated through MTT colorimetry and cell morphology. The antiviral activity of Ribavirin, Shuanghuanglian and Garlic were studied in HEL and Vero cells infected with CBV3 and ECHO11 by observing cytopathic effect (CPE), MTT colorimetry and plaque-reduction assay. The antiviral activity of these three drugs were compared and that of Shuanghuanglian and Garlic were also compared before and after ECHO11 absorbing by plaque-reduction assay. RESULTS: (1) The cytotoxicity of these three drugs were expressed as TC50(50% toxic concentration). TC50 of Ribavirin was 2 mg/ml, of Shuanghuanglian 5 mg/ml, of Garlic 12.5 micrograms/ml. (2) Ribavirin could inhibit CBV3 and ECHO11 at the concentration ranged from 1 mg/ml to 1.5 mg/ml. Shuanghuanglian could inhibit CBV3 and ECHO11 at the concentration of 0.5 mg/ml and the viral inhibiting effect was concentration-dependent. Garlic could inhibit CBV3 and ECHO11 at the concentration ranged from 2.5 micrograms/ml to 7.5 micrograms/ml and 5 micrograms/ml was the most effective. (3) Plaque-reduction assay was used to test the anti-virus (CBV3 and ECHO11) activity of these three drugs: plaque reduction rate of 1.5 mg/ml Ribavirin was 43.2% and 37.2%, of 2.5 mg/ml Shuanghuanglian was 81.1% and 88.4% and of 5 micrograms/ml Garlic was 66.2% and 77.4% respectively. The plaque reduction rate of Ribavirin was lower than the other two drugs (P < 0.05), between these two showed no significant difference (P > 0.05). The anti-ECHO11 activity of Shanghuanglian before ECHO11 adsorbing was higher than after ECHO11 adsorbing (P < 0.05). There was no significant difference between the plaque reduction rates of Garlic before and after ECHO11 adsorbin. CONCLUSION: All of the three drugs have anti-virus activity in vitro while Shuanghuanglian and Garlic are more effective. Among these three drugs, the cytotoxicity of Shuanghuanglian is the most weak and the anti-virus activity is the strongest. The antiviral activity of Shuanghuanglian adding before virus adsorbing was higher, so it seems that Shuanghuanglian can prevent EV infection.