4种清热解毒复方中草药体外抑菌及抗PRRSV活性研究

【目的】 探索泻心汤、三黄虎杖汤、黄连解毒汤和清瘟败毒散4种清热解毒复方中草药体外抑制大肠杆菌、金黄色葡萄球菌及抗猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus, PRRSV)的效果, 为临床应用提供参考。【方法】 采用传统水煎法对4种复方中草药进行提取, 通过二倍稀释法测定中药对大肠杆菌标准菌ATCC 259222和金黄色葡萄球菌标准菌ATCC 25923的体外抑菌活性; 在测定复方中草药对Marc-145细胞最大安全浓度的基础上, 通过实时荧光定量PCR鉴定复方中草药对PRRSV的抑制作用。【结果】 泻心汤对大肠杆菌的体外抑菌活性最好, 其最小抑菌浓度(minimum inhibitory concentration, MIC)和最小杀菌浓度(minimum bactericidal concentration, MBC)均为7.81 mg/mL; 三黄虎杖汤和黄连解毒汤次之, 其MIC和MBC为31.25~125 mg/mL; 清瘟败毒散抑制大肠杆菌效果较差: MIC和MBC均>250 mg/mL, 而金黄色葡萄球菌对泻心汤、黄连解毒汤和三黄虎杖汤非常敏感, 其MIC和MBC为0.24~0.98 mg/mL。在体外抗PRRSV作用的试验中, 泻心汤、三黄虎杖汤、黄连解毒汤和清瘟败毒散对Marc-145细胞最大安全浓度分别为0.49、0.49、3.90和1.95 mg/mL。4种复方中草药对PRRSV的抑制与直接灭活作用较好, 黄连解毒汤和清瘟败毒散均在0.98 mg/mL时对PRRSV有阻断作用, 泻心汤和三黄虎杖汤在0.24和0.12 mg/mL时对PRRSV有阻断效果。【结论】 4种复方中草药中泻心汤的体外抑菌及抗PRRSV的效果最好, 其次是三黄虎杖汤、黄连解毒汤和清瘟败毒散, 4种清热解毒复方均有一定的抑菌和抗病毒活性, 可以为临床应用提供参考依据。

Study on the Antibacterial and Anti-PRRSV Activities of Four Heat Clearing and Detoxifying Compound Chinese Herbal Medicine in vitro

【Objective】 The purpose of this study was to explore the effects of 4 heat clearing and detoxifying compound Chinese herbal medicines, Xiexin decoction, Sanhuang Huzhang decoction, Huanglian Jiedu decoction and Qingwen Baidu powder on inhibiting Escherichia coli and Staphylococcus aureus combined with anti Porcine reproductive and respiratory syndrome virus (PRRSV) in vitro, in order to provide reference to clinicuse.【Method】 Active ingredients of 4 kinds of compoud Chinese herbal medicines were extracted by the conventional decoction method.Double dilution method was used to determine the in vitro antibacterial activities of 4 kinds of Chinese herbal medicines and their compound against Escherichia coli standard strain ATCC 259222 and Staphylococcus aureus standard strain ATCC 25923.Based on the study of the maximum safe concentration of Chinese herbal medicine on Marc-145 cells, the inhibitory effect of compound traditional Chinese medicine on PRRSV was determined by Real-time quantitative PCR.【Result】 Xiexin decoction had the best antibacterial activity against Escherichia coli in vitro, its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were both 7.81 mg/mL, followed by Sanhuang Huzhang decoction and Huanglian Jiedu decoction, which MIC and MBC were 31.25-125 mg/mL.Qingwen Baidu powder had poor inhibitory effect on Escherichia coli, the MIC and MBC were more than 250 mg/mL, while Staphylococcus aureus was very sensitive to Xiexin decoction, Huanglian Jiedu decoction and Sanhuang Huzhang decoction.The MIC and MBC were 0.24-0.98 mg/mL.In the experiment of anti PRRSV effect in vitro, the maximum safe concentrations of Xiexin decoction, Sanhuang Huzhang decoction, Huanglian Jiedu decoction and Qingwen Baidu powder on Marc-145 cells were 0.49, 0.49, 3.90 and 1.95 mg/mL, respectively.4 compound Chinese herbal medicines had good inhibitory and direct inactivation effects on PRRSV.Huanglian Jiedu decoction and Qingwen Baidu powder had blocking effects on PRRSV at 0.98 mg/mL, while Xiexin decoction and Sanhuang Huzhang decoction had blocking effects on PRRSV at 0.24 and 0.12 mg/mL, respectively.【Conclusion】 It was suggested that among 4 compound Chinese herbal medicines, Xiexin decoction had the best antibacterial and anti PRRSV effects in vitro, followed by Sanhuang Huzhang decoction, Huanglian Jiedu decoction and Qingwen Baidu powder.4 heat clearing and detoxifying compounds had certain antibacterial and antiviral activities, which could provide reference basis for clinical application.