头花蓼对幽门螺杆菌粘附定植的影响

Influence of Polygonum capitatum on the adherency and colonization of Helicobacter pylori

目的观察头花蓼水提物对幽门螺杆菌(Helicobacter pylori,H.pylori)粘附定植的影响。方法采用尿素酶活性比色法定量检测头花蓼水提物作用前后幽门螺杆菌尿素酶活性的影响;运用半固体布氏琼脂平板法检测不同浓度头花蓼对幽门螺杆菌鞭毛运动的影响;利用细胞爬片革兰染色观察不同浓度头花蓼作用后幽门螺杆菌在GES-1细胞表面粘附的变化并计算各组抑制率;采用Real-time PCR技术检测头花蓼作用前后相关基因ureB、ureE、ureF、flaA、flaB、babA、alpA、alpB的mRNA转录水平。结果头花蓼水提物对幽门螺杆菌尿素酶活性具有抑制作用,抑制率为44.90±0.289。头花蓼浓度为1/2MIC、1/4MIC、1/8MIC分别作用后幽门螺杆菌菌膜晕圈直径显著小于对照组,分别为(5.67±0.55)mm,(8.5±0.50)mm,(11.67±1.53)mm,差异有统计学意义(P<0.05);不同浓度头花蓼水提物均能抑制细菌粘附胃上皮细胞,各浓度组与对照组比较,差异有统计学意义(P<0.05)。头花蓼作用后,与对照组比较,下调了尿素酶结构基因ureB,尿素酶活性基因ureE、ureF、鞭毛相关基因flaB、flaA以及粘附素基因babA、alpA、alpB mRNA的转录水平,差异均具有统计学意义(P<0.05)。结论头花蓼对幽门螺杆菌具有明显的抑制作用,是通过抑制幽门螺杆菌尿素酶活性,减缓幽门螺杆菌的鞭毛动力,影响幽门螺杆菌对胃粘膜上皮细胞的粘附等方面发挥其抑菌作用。

We aimed to observe the influence of P.capitatum on adherency and colonization of H.pylori.Urease active colorimetric method would be adopted to quantitatively detect the influence on urease activity of H.pylori before and after the function of P.capitatum;semi-solid Brucella agar plate method would be used to detect the influence on flagellar movement of H.pylori before and after the function of P.capitatum;Gram＇s stain of cell slides would be utilized to observe the changes of different concentrations of P.capitatumand H.pylori on the bacteria adhered on the surface of GES-1cells and calculate the inhibition ratio of all groups;real-time PCR technologies would be adopted to detect the expression level of related genes of ureE,ureF,flaA,flab,babA,alpAand alpBbefore and after the function of P.capitatum.P.capitatum had inhibition effect on urease activity of H.pylori with the inhibition ratio being 44.90±0.289.The concentration of P.capitatum was respectively 1/2MIC,1/4MIC and 1/8MIC;after respective function,the diameter of H.pylori pellicle halo was significantly smaller than that of the control group,respectively（5.67±0.55）mm,（8.5±0.50）mm and（11.67±1.53）mm with statistical difference（P〈0.05）;different concentrations of P.capitatumcould all inhibit the bacteria to adhere to gastric epithelial cells and compared to the control groups,all concentration groups were with statistical difference（P〈0.05）.After the function of P.capitatum,compared to the control group,the expression level of urease active genes ureEand ureF,flagellin genes flab and flaA as well as adhesion genes babA,alpA and alpB mRNA was reduced with statistical difference（P〈0.05）.It came to the conclusion that P.capitatumhas obvious inhibition effect on H.pylori,which exerts its antibacterial effect via inhibiting urease activity of H.pylori,slowing down the flagella movement of H.pylori and impacting the adherence of H.pylori to gastric epithelial cells.