广藿香醇抗幽门螺杆菌的作用及对肠道微生物的影响

Anti-Helicobacter pylori Effects of Patchouli Aalcohol and Its Influences on Intestinal Microbiota during Treatment

目的探讨广藿香醇(Patchouli alcohol,PA)抗幽门螺杆菌(Helicobacter pylori)的作用及其治疗过程中对肠道微生物的影响。方法于Brain-Heart Infusion(BHI)培养液中加入PA,使其终浓度为50、25、12.5、6.25μg·mL-1,液体稀释法测定PA对幽门螺杆菌3株标准株及2株临床耐药株(甲硝唑及克拉霉素耐药)的最小抑菌浓度(MIC)和最小杀菌浓度(MBC);选择浓度为1/4 MIC、1/2 MIC的PA,荧光法测定PA抗幽门螺杆菌NCTC11637菌株黏附细胞的作用。将雄性C57BL/6小鼠随机分为阴性对照组(泊洛沙姆)、阳性对照组(抗生素三联疗法)及PA高(20 mg·kg-1)、低(5 mg·kg-1)剂量组,给药14 d。二代测序法测PA对肠道微生物菌群的影响,同时用RT-qPCR法测肠道菌群中大肠杆菌和双歧杆菌的丰度变化。结果 PA抗幽门螺杆菌临床株及标准株的MIC值为6.25~12.5μg·mL-1,MBC值为12.5~50μg·mL-1;1/2和1/4 MIC的PA可以降低幽门螺杆菌的黏附能力;抗生素三联疗法可显著性降低小鼠肠道微生物多样性及丰度,影响肠道菌群中两个主要菌群所占比例,而PA对肠道微生物菌群无明显影响。结论 PA具有抗幽门螺杆菌作用,能够降低幽门螺杆菌的黏附能力,在治疗幽门螺杆菌感染的同时对肠道微生物菌群影响较小。

Objective In order to clarify the therapeutic advantages of Patchouli alcohol(PA)in treatment of Helicobacter pylori infection,experiments were carried out to investigate the anti-H.pylori effects of PA,and its possible influences on intestinal microbiota during treatment.Methods PA was added to Brain-Heart Infusion(BHI)medium at final concentrations of 50,25,12.5 and 6.25μg·mL-1.Then,the broth dilution method was used to measure the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of PA against three standard and two clinical drug-resistant(Metronidazole and clarithromycin-resistance strains)H.pylori strains.Based on the MIC value,1/4 MIC and 1/2 MIC of PA were selected to explore the anti-H.pylori adhesion capacity using fluorescence method.Besides,20 male C57 BL/6 mice were randomly divided into control,positive,PA high dose and low dose groups.Positive group mice were treated with triple therapy,while the PA and control group mice were orally administrated with corresponding PA and poloxamer,respectively.After 14 days,the influences of PA on mice intestinal microbiota were investigated by next generation sequencing technology.Meanwhile,the abundance of the two major bacterial groups(Bifidobacterium and Escherichia coli)in the intestinal flora were determined by RT-qPCR.Results The MIC values of PA against H.pylori clinical drug-resistant and standard strains were 6.25-12.5μg·mL-1,MBC values were 12.5-50μg·mL-1;1/2 and 1/4 MIC PA could reduce the adhesion ability of H.pylori.Triple therapy can significantly reduce the diversity and abundance of mice intestinal microbiota,and affect the proportion of those two major intestinal flora,while no remarkable influence of PA on intestinal microflora was observed.Conclusion The anti-H.pylori capacity of PA was shown.Reliable inhibition on the H.pylori adhesion ability of PA was also presented.Besides,less influence of PA on mice intestinal microbiota was observed during the treatment.