50种中药甲醇提取物对嗜水气单胞菌的体外抑菌及抗生物膜活性的评价

In vitro antibacterial and anti-biofilm activities of 50 kinds of traditional Chinese medicine methanol extracts against Aeromonas hydrophila

目的 评价50种中药甲醇提取物对嗜水气单胞菌的体外抑菌效果以及抗生物膜生成活性。方法 采取甲醇浸提的方法制备中药甲醇提取物,计算提取率;利用多功能酶标仪检测600 nm处的吸光度(A600)值测定各提取物(0.1、0.2、0.4、0.8、1.6、3.2 mg·mL^(-1))作用16 h对嗜水气单胞菌的抑菌率;结晶紫染色法结合多功能酶标仪测定各提取物的抗生物膜活性。结果 苏木、白芍、侧柏叶等32种中药甲醇提取物对嗜水气单胞菌抑菌效果明显,其中苏木甲醇提取物的抑菌效果最显著,最大抑菌率达95.56%,起效质量浓度为0.1 mg·mL^(-1);石菖蒲、大青叶、广藿香等20种中药甲醇提取物明显抑制嗜水气单胞菌生物被膜的生成,其中白芍的最大抑制率达96.78%。结论 夏枯草、野菊花、大青叶、赤芍、石菖蒲、地肤子、连翘、苏木、肉桂、女贞子、白芍、甘草、肉豆蔻、鸡血藤、乌药、莪术、侧柏叶17种中药对嗜水气单胞菌同时具有抑菌和抑制生物膜生长活性。

Objective To evaluate the in vitro antibacterial and anti-biofilm activities of 50 kinds of tranditional Chiese medicine(TCM) extracts against Aeromonas hydrophila. Methods The methanol extract was prepared by methanol extraction and the extraction rate was calculated. Multifunctional enzyme marker was used to detect the absorbance(A600 nm) value at 600 nm to determine the inhibitory rate of each extract(0.1, 0.2, 0.4, 0.8, 1.6, 3.2 mg·mL^(-1)) against A. hydrophila for 16 h. The antibiofilm activity of the extracts was determined by crystal violet staining and multifunctional enzyme marker. Results Totally 32 kinds of TCM methanol extracts, including Caesalpinia sappan, Paeonia iactiflora and Platycladus orientalis and so on, showed obvious bacteriostatic effect on A. hydrophila, among which Sappan Lignum showed the most significant bacteriostatic effect with the maximum bacteriostatic rate of 95.56% and effective concentration of 0.1 mg·mL^(-1). The methanol extracts of 20 kinds of TCM, such as Acorus tatarinowii, Isatis indigotica and Pogostemon cablin and so on, inhibited the biofilm formation of A. hydropaeoniae, and the maximum inhibitory rate of Paeonia iactiflora was 96.78%. Conclusion 17 TCM have bothe antibacterial and anti-biofilm activity, including Prunella vulgaris, Chrysanthemum indicum, Isatis indigotica, Paeonia iactiflora, Acorus tatarinowii, Kochia scoparia, Forsythia suspensa, Caesalpinia sappan, Cinnamomum cassia, Ligustrum lucidum, Paeonia iactiflora, Glycyrrhiza uralensis, Myristica fragrans, Spatholobus suberectus, Lindera aggregata, Curcuma phaeocaulis, Platycladus orientalis.