氢溴酸槟榔碱对幽门螺杆菌生长和空泡毒素A表达和活性的影响

Effect of arecoline hydrobromide on H. pylori growth and expression and activity of vacuolating cytotoxin A

目的评价氢溴酸槟榔碱对幽门螺杆菌(Helicobacterpylori,H.pylori)的抑制作用及其对H.pylori空泡毒素A表达水平和活性的影响。方法使用平皿稀释固体法和脑心浸液液体法测定中药活性成分氢溴酸槟榔碱对H.pylori的最低抑菌浓度(MIC),以反映药物对H.pylori的抑制作用。通过中性红摄入法评价药物干预后H.pylori培养上清中空泡毒素A对胃癌细胞BGC-823的毒性作用,并通过RT-PCR方法和Western blot方法检测经药物处理后H.pylori菌体及分泌上清中空泡毒素A mRNA和蛋白的表达水平,以反映药物对H.pylori空泡毒素A活性和表达的影响。结果高浓度氢溴酸槟榔碱对H.pylori具有抑制作用,MIC为:固体法和液体法均为1.25 g·L-1。低浓度氢溴酸槟榔碱可不同程度抑制H.pylori空泡毒素A的活性(P<0.01);抑制H.pylori菌体和分泌上清中空泡毒素A mRNA(P<0.05)和蛋白(P<0.05)的表达。结论氢溴酸槟榔碱在较高浓度(1.25 g·L-1)时具有抑制H.pylori生长作用,而在较低浓度(0.15 g·L-1)时,即可抑制H.pylori空泡毒素A的表达和活性。

Aim To investigate the effect of arecoline hydrobromide （AH） , a Chinese herbal active ingredient, on H. pylori growth and the expression and activity of vacuolating cytotoxin A （VacA）. Methods The minimum inhibitory concentration （MIC） of AH to H. pylori was detected using the diluted solid Columbia agar and the brain-heart extract liquid medium separately to illustrate the inhibitory effect of AH on H. pylori. To identify the effect of AH on the activity and expression of VaeA in H. pylori, the toxic effect of VaeA secreted from AH-treated H. pylori on BGC-823, a gastric cancer cell, was identified by neutral red uptake test （NRU）. The mRNA and protein levels of vacA in AH-treated H. pylori were detected by RT-PCR and Western blot, respectively. Results MICs of AH to H. pylori were both 1.25 g ·L-1 either on solid agar or in liquid medium. Compared with the control, AH could inhibit the VacA activity （ P 〈 0. 05 ） and the mRNA and protein expression of VacA （ P 〈 0. 05, P 〈 0. 05 ）. Conclusion AH of high concentration （ 1.25 g · L-1 ） can inhibit the growth of H. pylori , while that of lower level（0. 15 g · L-I） can inhibit the VacA activity and expression in vitro.