山橿抗Hp有效组分筛选及其对Hp感染胃癌AGS细胞的影响

Screening of anti-Helicobacter pylori active components from Lindera reflexa Hemsl.and their effects on Helicobacter pylori infection of AGS cells

目的探究山橿乙酸乙酯部位及其化学成分对幽门螺杆菌(Helicobacter pylori,Hp)的体外抑菌活性,筛选抗Hp有效组分;进一步研究有效组分对Hp感染人胃癌AGS细胞的影响。方法药敏纸片法比较山橿乙酸乙酯部位及其化学成分对Hp的体外抑菌效果,筛选山橿乙酸乙酯部位抗Hp有效成分,组建有效组分;对有效组分及抗Hp最强的化合物,以标准菌株Hp P12、三株多重耐药Hp菌株为实验菌株,克拉霉素为阳性对照药,采用倍比稀释法测定其最低抑菌浓度(MIC)值;细胞实验检测二者对人胃癌AGS细胞增殖、凋亡,Hp感染AGS细胞黏附能力的影响。结果山橿乙酸乙酯部位中二苯乙烯类成分的抗Hp作用优于黄酮类成分,其中银松素的抗Hp作用最为显著;依据山橿乙酸乙酯部位中8个二苯乙烯类成分的含量组建山橿抗Hp有效组分;山橿抗Hp有效组分对Hp标准菌株的MIC值为12~16μg/mL,对Hp耐药菌株的MIC值为256~512μg/mL;银松素对Hp标准菌株的MIC值为16~32μg/mL,对Hp耐药菌株的MIC值为64~128μg/mL,均明显优于山橿乙酸乙酯部位。山橿抗Hp有效组分、银松素对AGS细胞的IC50分别为14.10μg/mL,59.75μg/mL,且二者均不影响AGS细胞凋亡,又可在一定程度上减少Hp的定植。结论山橿乙酸乙酯部位抗Hp的主要有效成分为其中的二苯乙烯类化合物,其中以银松素的作用最强,其抗Hp机制与减少Hp的定植有关。

Objective To investigate and screen the anti-Helicobacter pylori(anti-Hp)activity of chemical components from the ethyl acetate effective fraction of Lindera reflexa Hemsl.in vitro,and then further study the effect of active components on human gastric cancer AGS cells infected by Hp.Methods The anti-Hp effect of the ethyl acetate effective fraction from Lindera reflexa Hemsl.and its chemical components were compared by Kirby-Bauer disk diffusion method,and the anti-Hp effective components were screened to construct the anti-Hp effective component.The MIC values of the anti-Hp effective component and effective component with best antibacterial activity were determined by multiple dilution method,using standard strain Hp P12 and three multi-drug resistant strains Hp as the experimental strains and Clarithromycin as the control drug.Subsequently,cell experiments were used to detect the effects of both on the proliferation and apoptosis of human gastric cancer AGS cells,and the adhesion ability on human gastric cancer AGS cells infected by Hp.Results The anti-Hp effect of the stilbene components in the ethyl acetate effective fraction of Lindera reflexa Hemsl.was better than that of the flavonoid components,with pinosylvin having the most significant anti-Hp effect.Based on the content of 8 stilbene components in the ethyl acetate effective fraction,the anti-Hp effective component were established.The MIC value of the anti-Hp effective component against Hp standard strain was 12~16μg/mL,with a MIC value of 256~512μg/mL for Hp resistant strains.The MIC value of pinosylvin against Hp standard strain was 16~32μg/mL,with a MIC value of 64~128μg/mL for Hp resistant strains,all of them were significantly better than that of the ethyl acetate fraction of Lindera reflexa Hemsl..The IC50 of the anti-Hp effective component and pinosylvin on AGS cells were 14.10μg/mL and 59.75μg/mL,respectively.Both of them did not affect the apoptosis of AGS cells,and could reduce the colonization of Hp to a certain extent.Conclusion The main effective components of the ethyl acetate fraction from Lindera reflexa Hemsl.inhibited Hp are its stilbene compounds,among which pinosylvin has the strongest effect.Their anti-Hp mechanism is related to reducing the colonization of Hp.