蒲公英水提物对临床常见肠道有害菌体外抑菌作用的研究

In Vitro Antibacterial Effect of Dandelion Aqueous Extract on Clinical Intestinal Harmful Bacteria

目的研究中草药蒲公英水提物对临床常见肠道有害菌体外抑菌作用。方法配制不同浓度的蒲公英药液:1.0、0.5、0.25、0.125、0.06、0.03、0 g/ml,分别作用于产气荚膜梭菌、脆弱拟杆菌及蜡样芽胞杆菌3种肠道有害菌,将干预后的细菌分别采用连续划线方法接种于血琼脂平皿,培养24 h后通过计数菌落数判断3种肠道有害菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC);采用甲酸萃取法提取细菌样本,利用基质辅助激光解析串联飞行时间质谱仪(MALDI-TOF)FlexControl 2.0软件检测样品后生成图谱,应用FlexAnalysis 3.0软件分析图谱及参数,判断蒲公英水提物对肠道有害菌蛋白种类及含量的影响。结果蒲公英对产气荚膜梭菌的MIC为0.06 g/ml,MBC为0.25 g/ml;对脆弱拟杆菌的MIC为0.03 g/ml,MBC为0.125 g/ml;对蜡样芽胞杆菌的MIC为0.125 g/ml,MBC为0.5 g/ml;蒲公英药物浓度为0、0.03、0.06、0.125 g/ml时,产气荚膜梭菌蛋白种类离子m/z种类分别为35、8、6、6种;蒲公英药物浓度为0、0.03、0.06 g/ml时,脆弱拟杆菌蛋白种类离子m/z数量种类分别为76、47、44种;蒲公英药物浓度为0、0.03、0.06、0.125、0.25 g/ml时,蜡样芽胞杆菌蛋白种类离子m/z种类分别为40、42、32、7、5种。结论中草药蒲公英的水提物对临床常见的肠道有害菌的生长及蛋白合成具有很好的抑制作用。

Objective To study the in vitro antibacterial effect of dandelion water extract on common intestinal harmful bacteria.Methods Different concentrations of dandelion solution were prepared:1.0,0.5,0.25,0.125,0.06,0.03,0 g/ml,respectively,to stimulate Clostridium perfringens,Bacteroides fragilis and Bacillus cereus.After the intervention,the bacteria were inoculated into the blood agar plate by continuous scribing method,and the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of the three intestinal harmful bacteria were determined by counting the number of colonies after 24 hours of culture.The formic acid extraction method was used to extract the bacterial samples.The matrix assisted laser desorption tandem time of flight mass spectrometer(MALDI-TOF)FlexControl 2.0 software was used to detect the samples and generate the atlas.The FlexAnalysis 3.0 software was used to analyze the atlas and parameters to determine the impact of the aqueous extract of dandelion on the type and content of intestinal harmful bacteria protein.Results The MIC and MBC of dandelion to Clostridium perfringens was 0.06 g/ml and 0.25 g/ml,Bacteroides fragilis was 0.03 g/ml and 0.125 g/ml,Bacillus cereus was 0.125 g/ml and 0.5 g/ml.When the concentration of dandelion was 0,0.03,0.06,0.125 g/mL,the type ions m/z of protein of Clostridium perfringens were 35,8,6 and 6.When the concentration of dandelion was 0,0.03,0.06 g/mL,the number and types of the type ions m/z of the fragile Bacteroides protein were 76,47,and 44.When the concentration of dandelion was 0,0.03,0.06,0.125 and 0.25 g/ml,the type ions of Bacillus cereus protein m/z were 40,42,32,7 and 5.Conclusion The aqueous extract of dandelion has different degrees of inhibition on the growth and protein synthesis of intestinal harmful bacteria.