广藿香醇抑制幽门螺杆菌脲酶活性及其机制

Mechanism of anti-Helicobacter pylori urease activity of patchouli alcohol

探讨广藿香醇对于幽门螺杆菌Helicobacter pylori脲酶活性的抑制及其相关基因的表达变化,为进一步研究广藿香醇对幽门螺杆菌定植感染的影响奠定基础。培养幽门螺杆菌并采用革兰染色、快速尿素酶及PCR的方法鉴定后,在酸性条件(pH 5.3)和中性条件(pH 7.0)培养液中给予不同浓度的广藿香醇干预1 h,应用琼脂稀释法测定细菌的存活率;Berthelot显色法检测细菌的脲酶活性;RT-qPCR法检测细菌中ureA,ureB,ureE,ureH,ureI和nix A相关脲酶基因的表达变化。经过鉴定生长良好幽门螺杆菌在不同浓度的广藿香醇干预后,细菌存活率没有明显变化;其脲酶活性出现了显著的下降;同时脲酶的相关基因ureA,ureB,ureE,ureH,ureI和nix A的表达均出现不同程度的降低。广藿香醇在酸性条件和中性条件下均可以抑制幽门螺杆菌脲酶的活性,可能与降低细菌脲酶相关的基因表达有关。

To investigate the effect of patchouli alcohol on inhibiting Helicobater pylori urease activity,and its effect on expression levels of related genes,and lay the foundation for further research on the effect of patchouli alcohol on H. pylori colonization and infection. H. pyloriwas cultured and identified by gram staining,rapid urease test（ RUT） and PCR method. Then agar dilution method was used to detect the bacterial survival after 1 h intervention by different concentrations of patchouli alcoholin the acidic（ pH 5. 3） and neutral（ pH 7. 0） conditions; berthelot method was used to detect urease activity and RT-qPCR method was used to detect the expression changes of ureA,ureB,ureE,ureH,ureI,and nix A related urease genes. The results showed that the survival rate of H. pyloriwas not significantly changed but the urease activity was obviously decreased after intervention by different concentrations of patchouli alcohol; meanwhile,the expression levels of ureA,ureB,ureE,ureH,ureI,and nix A were decreased to different degrees. Therefore,patchouli alcohol could inhibit H. pylori urease activity in both acidic and neutral conditions,and the mechanism may be related to down-regulation of urease gene expression.