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.
[1] MALFERTHEINER P, MEGRAUD F, O′MORAIN C A, et al.Management of Helicobacter pylori infection-the Maastricht V/florence consensus report[J].Gut, 2017, 66(1):6-30.
[2] SUGANO K, TACK J, KUIPERS E J, et al.Kyoto global consensus report on Helicobacter pylori gastritis[J].Gut, 2015, 64(9):1353-1367.
[3] EL-SERAG H B, KAO J Y, KANWAL F, et al.Houston consensus conference on testing for Helicobacter pylori infection in the United States[J].Clinical Gastroenterology and Hepatology, 2018, 16(7):992-1002.
[4] WONG B C Y, LAM S K, WONG W M, et al.Helicobacter pylori eradication to prevent gastric cancer in a high-risk region of China:A randomized controlled trial[J].JAMA, 2004, 291(2):187-194.
[5] ZHOU X Z, LYU N H, ZHU H Y, et al.Large-scale, national, family-based epidemiological study on Helicobacter pylori infection in China:The time to change practice for related disease prevention[J].Gut, 2023, 72(5):855-869.
[6] 中华医学会消化病学分会幽门螺杆菌学组. 第六次全国幽门螺杆菌感染处理共识报告(非根除治疗部分)[J].胃肠病学, 2022, 27(5):289-304.
Helicobacter pylori Study Group, Chinese Society of Gastroenterology, Chinese Medical Association.Sixth Chinese national consensus report on the management of Helicobacter pylori infection (treatment excluded)[J].Chinese Journal of Gastroenterology, 2022, 27(5):289-304.
[7] ROSZCZENKO-JASIŃSKA P, WOJTYŚ M I, JAGUSZTYN-KRYNICKA E K.Helicobacter pylori treatment in the post-antibiotics era-searching for new drug targets[J].Applied Microbiology and Biotechnology, 2020, 104(23):9891-9905.
[8] GUPTA M, DEY S, MARBANIANG D, et al.Grape seed extract:Having a potential health benefits[J].Journal of Food Science and Technology, 2020:57(4):1205-1215.
[9] SILVAN J M, GUTIÉRREZ-DOCIO A, MORENO-FERNANDEZ S, et al.Procyanidin-rich extract from grape seeds as a putative tool against Helicobacter pylori[J].Foods, 2020, 9(10):1370.
[10] HŘIBOVÁ P, KHAZNEH E, ŽEMLIČKA M, et al.Antiurease activity of plants growing in the Czech Republic[J].Natural Product Research, 2014, 28(12):868-873.
[11] CHUA E G, VERBRUGGHE P, PERKINS T T, et al.Fucoidans disrupt adherence of Helicobacter pylori to AGS cells in vitro[J].Evidence-Based Complementary and Alternative Medicine, 2015, 2015:120981.
[12] APOSTOLOVA E, LUKOVA P, BALDZHIEVA A, et al.Immunomodulatory and anti-inflammatory effects of fucoidan:A review[J].Polymers, 2020, 12(10):2338.
[13] NI L Y, WANG L, FU X T, et al.In vitro and in vivo anti-inflammatory activities of a fucose-rich fucoidan isolated from Saccharina japonica[J].International Journal of Biological Macromolecules, 2020, 156:717-729.
[14] SHEN X, ZHANG W J, PENG C, et al.In vitro anti-bacterial activity and network pharmacology analysis of Sanguisorba officinalis L. against Helicobacter pylori infection[J].Chinese Medicine, 2021, 16(1):33.
[15] YAN J H, PENG C, CHEN P T, et al.In-vitro anti-Helicobacter pylori activity and preliminary mechanism of action of Canarium album Raeusch.fruit extracts[J].Journal of Ethnopharmacology, 2022, 283:114578.
[16] 张姝, 罗昭逊, 莫非, 等.头花蓼对幽门螺杆菌粘附定植的影响[J].中国人兽共患病学报, 2016, 32(8):734-740.
ZHANG S, LUO Z X, MO F, et al.Influence of Polygonum capitatum on the adherency and colonization of Helicobacter pylori[J].Chinese Journal of Zoonoses, 2016, 32(8):734-740.
[17] XU Y F, LIAN D W, CHEN Y Q, et al.In vitro and in vivo antibacterial activities of patchouli alcohol, a naturally occurring tricyclic sesquiterpene, against Helicobacter pylori infection[J].Antimicrobial Agents and Chemotherapy, 2017, 61(6):e00122-17.
[18] YANG H, HUANG X X, ZHANG X C, et al.AI-2 induces urease expression through downregulation of orphan response regulator HP1021 in Helicobacter pylori[J].Frontiers in Medicine, 2022, 9:790994.
[19] SHARNDAMA H C, MBA I E.Helicobacter pylori:An up-to-date overview on the virulence and pathogenesis mechanisms[J].Brazilian Journal of Microbiology, 2022, 53(1):33-50.
[20] 许艺飞. 广藿香醇抗幽门螺杆菌作用及其机制研究[D].广州:广州中医药大学, 2016.
XU Y F.Anti-Helicobacter pylori effect and mechanism of patchouli alcohol[D].Guangzhou:Guangzhou University of Chinese Medicine, 2016.
[21] SENKOVICH O A, YIN J, EKSHYYAN V, et al.Helicobacter pylori AlpA and AlpB bind host laminin and influence gastric inflammation in gerbils[J].Infection and Immunity, 2011, 79(8):3106-3116.
[22] MATOS R, AMORIM I, MAGALHÃES A, et al.Adhesion of Helicobacter species to the human gastric mucosa:A deep look into glycans role[J].Frontiers in Molecular Biosciences, 2021, 8:656439.
[23] JOSENHANS C, LABIGNE A, SUERBAUM S.Comparative ultrastructural and functional studies of Helicobacter pylori and Helicobacter mustelae flagellin mutants:Both flagellin subunits, FlaA and FlaB, are necessary for full motility in Helicobacter species[J].Journal of Bacteriology, 1995, 177(11):3010-3020.
[24] 罗微, 王倩, 陈铁龙.幽门螺杆菌鞭毛研究概述[J].中国病原生物学杂志, 2015, 10(9):852-857.
LUO W, WANG Q, CHEN T L.Advances in the study of Helicobacter pylori flagella[J].Journal of Pathogen Biology, 2015, 10(9):852-857.
[25] PALFRAMAN S L, KWOK T, GABRIEL K.Vacuolating cytotoxin A (VacA), a key toxin for Helicobacter pylori pathogenesis[J].Frontiers in Cellular and Infection Microbiology, 2012, 2:92.
