土荆芥提取物在体外对幽门螺杆菌生物膜的作用

Effect of Chenopodium ambrosioides L.on Biofilm Formation of Helicobacter pylori in Vitro

目的研究土荆芥提取物在体外对幽门螺杆菌(Hp)生物膜形成的作用。方法建立Hp菌株(NCTC11637、26995-16R)体外生物膜模型,选取稳定成膜的菌株,E-test法鉴定菌株的耐药能力,琼脂稀释法测量土荆芥提取物(挥发油)的最低抑菌浓度(MIC),采用96孔板结晶紫染色法测定D(OD均值),扫描电镜观察不同浓度的土荆芥油及其联合甲硝唑时对Hp生物膜形成的影响。结果甲硝唑耐药菌26995-16R为强生物膜Hp菌株,土荆芥MIC值为256μg/mL,D加药<D阴性、D土荆芥+甲硝唑<D甲硝唑,土荆芥组D值在一定范围内(≤1/2MIC)随着土荆芥浓度的增高而降低,扫描电镜下土荆芥对Hp的生物膜结构破坏作用及杀菌作用随着药物浓度的增加而增加,土荆芥联合甲硝唑比单独使用甲硝唑的作用更强。结论土荆芥提取物在体外对于Hp菌株的生物膜形成具有抑制作用,联合抗生素时具有协同抑制生物膜形成的作用。

Objective To observe the effect of Chenopodium ambrosioides L.(CAL)on biofilm formation of H.pylori in vitro.Methods The models of biofilm formation of H.pylori(NCTC11637,26995-16 R)were established in vitro and the strain with a strong biofilm-forming ability was chosen.Antibiotic resistance was determined by E-test.The minimal inhibitory concentration(MIC)of CAL was tested by agar dilution method.The 96-well plate model and crystal violet staining were used to determine the biofilm biomass(optical density,OD value),and D was the average value of OD.The effects of CAL with different concentrations and combinations with metronidazole(MTZ)on biofilm formation of H.pylori in vitro were observed by scanning electron microscope(SEM).Results H.pylori metronidazole resistant strain named 26995-16 R showed strong biofilm-forming ability.The MIC of CAL against 26995-16 R was about 256μg/mL.The D values of the drug groups were both lower than the negative control group.The D value of the group with both CAL and MTZ put was lower than that of the MTZ group.The D values of the CAL groups decreased with increasing drug concentrations under certain circumstance(≤1/2 MIC).Biofilm formation was destroyed more and bacteria were killed more along with increasing CAL concentrations in the CAL groups under SEM.CAL and MTZ had a synergistic effect in destroying biofilm formation and killing bacteria under SEM.Conclusion CAL had an inhibiting effect of biofilm formation of H.pylori in vitro,as well as their synergistic effect in combination with antibiotics.