葡萄籽提取物和岩藻多糖组合物抗幽门螺杆菌作用及机制研究  

Antimicrobial effects and mechanisms of grape seed extract and fucoidan combination on Helicobacter pylori

作　　者：孙静 邹圆静 孙璇 郑嘉妮 余丹阳 黄桂霞 姚美村 陈朋 SUN Jing;ZOU Yuanjing;SUN Xuan;ZHENG Jiani;YU Danyang;HUANG Guixia;YAO Meicun;CHEN Peng(Sirio Pharma Co.Ltd.,Shantou 515000,China;School of Pharmaceutical Sciences(Shenzhen),Sun Yat-sen University,Shenzhen 510817,China)

机构地区：[1]仙乐健康科技股份有限公司,广东汕头515000 [2]中山大学药学院(深圳),广东深圳510817

出　　处：《食品与发酵工业》2024年第18期279-283,共5页Food and Fermentation Industries

摘　　要：该研究旨在探索葡萄籽提取物和岩藻多糖组合物体外抗幽门螺杆菌及其机制。该实验在体外培养幽门标准菌株ATCC 700392,采用微量肉汤稀释法测定组合物的最小抑菌浓度(minimum inhibitory concentration,MIC)及时间-抑菌曲线;采用实时荧光定量(real-time fluorescence quantitative PCR,RT-qPCR)方法测定组合物对脲酶相关ureB、ureI、鞭毛相关flaB、flgK、黏附相关alpA、babA、毒力相关VacA基因表达水平的影响;采用扫描电镜观察组合物对菌体形态的影响。结果显示,组合物对幽门标准菌株ATCC 700392具有一定的体外抗菌活性,其MIC为1.5 g/L。在抑制动力学中,组合物在0.375 g/L时,ATCC 700392菌株的生长开始受到抑制,抑制活性呈现剂量依赖,浓度越大则抑制效果越明显,当组合物质量浓度为1.5 g/L时可完全抑制幽门螺杆菌的生长。RT-qPCR检测结果显示,组合物可显著抑制脲酶相关ureI、鞭毛相关flaB、flgK、黏附相关alpA、babA、毒力相关VacA基因等的表达。此外,扫描电镜观察到经组合物处理的菌体表面出现破损、菌体凹陷萎缩等明显的形态破损特征。该研究结果表明葡萄籽和岩藻多糖组合物在体外对幽门螺杆菌标准菌株有明显的抑制活性,其作用机制可能是通过抑制脲酶、抑制鞭毛相关的运动能力、抗黏附、破坏细菌形态等起到抑制幽门螺杆菌定殖的作用。该研究为更好地开发和利用葡萄籽岩藻多糖组合物提供了良好的科学实验支撑。This study aims to explore the in vitro antimicrobial effects and mechanisms of the combination of grape seed extract and fucoidan on Helicobacter pylori.In this experiment,the minimum inhibitory concentration(MIC)and inhibiting kinetic curve of the combination were determined using the microdilution method against the H.pylori standard strain ATCC 700392.The impacts of the combination on the expression levels of urease-related ureB and ureI,flagella-related flaB and flgK,adhesion-related alpA and babA,as well as the virulence-related VacA genes were assessed using RT-qPCR.Additionally,the effect of the combination on bacterial morphology was evaluate through electron microscopic scanning.Results of the study revealed that the combination exhibited significant antimicrobial activity against the H.pylori standard strain ATCC 700392,with an MIC of 1.5 g/L.In the inhibition kinetics,the combination was able to inhibit the growth of ATCC 700392 strain in a dose-dependent manner in a range from 0.375 g/L to 1.5 g/L.RT-qPCR results demonstrated that the combination significantly suppressed the expression of ureI,flaB and flgK,alpA and babA,as well as VacA genes.Moreover,scanning electron microscopy revealed that the combination changed the morphological integrity of the bacteria,including causing their surface damage and shrinkage.These findings suggest that the combination of grape seed extract and fucoidan exhibits significant inhibition of H.pylori standard strain in vitro,and its mechanism of action(MOA)may involve the inhibition of urease,suppression of flagella-related motility,anti-pathogenic adhesion,and disruption of bacterial morphology,all reducing the viability and colonizability of H.pylori.This research provides promising scientific support for the further development and utilization of grape seed and fucoidan combinations.

关 键 词：葡萄籽提取物 岩藻多糖 幽门螺杆菌 抗菌活性 作用机制 

分 类 号：TS209[轻工技术与工程—食品科学]