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目的:观察黄芪含药血清能否有效抑制体外幽门螺杆菌(hel i cobact er pyl ori,Hp)空泡毒素(VacA)的活性,从而探讨其抗菌机制。方法:将HP菌株培养上清液制成粗制VacA蛋白,经倍比稀释后(1∶32～1∶1),选择1∶2滴度与不同浓度黄芪含药血清共同作用于胃癌SGC-7901细胞,观察胃癌细胞形态变化,采用中性红摄入法(NRU)评价空泡活性。结果:黄芪含药血清对VacA活性有显著抑制作用,含药血清小剂量组NRU检测其空泡活性A550=0.228±0.029,显微镜下计数空泡形成率为78.75%±8.539%,与阴性对照组比较差异有显著性(P<0.05),随血清含药浓度的增加A550和空泡形成率逐渐降低;含药血清大剂量组A550=0.127±0.047,空泡形成率为32.25%±10.21%,与阴性对照组比较差异有极显著性(P<0.01),与阳性对照奥美拉唑组比较差异无显著性(P>0.05);含药血清大剂量组与中剂量组A550比较,差异无显著性(P>0.05),与小剂量组比较差异有显著性(P<0.05),可见黄芪含药血清对空泡毒素的抑制作用具有浓度依赖性。结论:黄芪对HP VacA活性具有明显的抑制作用,且该抑制作用具有一定的浓度依赖性。
Objective: To observe whether Huangqi medicine-containing serum can effectively inhibit the activity of vacuolar toxin (VacA) of Helicobacter pylori (Hp) in vitro and explore its antibacterial mechanism. Methods: The crude VacA protein was prepared from the culture supernatant of HP strains. After diluted 1:2 to 1:1, 1:2 titer and different concentrations of Astragalus membranaceus serum were used to treat gastric cancer SGC- 7901 cells were observed for morphological changes of gastric cancer cells, and vacuolar activity was evaluated by Neutral Red Intake (NRU). RESULTS: The serum containing Astragalus membranaceus significantly inhibited the activity of VacA. The vacuolating activity of A549 in the low-dose group of drug-containing serum was 0.258±0.029. The microscopic vacuolization rate was 78.75%±8.539%. The difference was statistically significant (P<0.05). With the increase of serum drug concentration, A550 and vacuolization rate gradually decreased; A550 with large dose of drug serum was 0.127±0.047, and the vacuolization rate was 32.25%±10.21%. Compared with the negative control group, the difference was extremely significant (P<0.01), and there was no significant difference compared with the positive control omeprazole group (P>0.05). The difference between the high-dose group and the middle-dose group A550 was not significant. Significant (P>0.05), compared with the low-dose group, the difference was statistically significant (P<0.05). The inhibitory effect of Huangqi-containing serum on vacuolating toxin was concentration-dependent. Conclusion: Astragalus membranaceus has a significant inhibitory effect on the activity of HP VacA, and this inhibition has a certain concentration dependence.