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过氧化氢建立的高氧环境对幽门螺杆菌清除作用的研究

Oxygen-enriched Environment Established by Hydrogen Peroxide Eradicates Helicobacter pylori in vitro and in vivo

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【作者】 狄佳张军屈伟程妍常丹燕郑向红

【Author】 DI Jia;ZHANG Jun;QU Wei;CHENG Yan;CHANG Danyan;ZHENG Xianghong;Department of Nuclear Medicine,the Second Affiliated Hospital of Xi’an Jiaotong University;Department of Gastroenterology,the Second Affiliated Hospital of Xi’an Jiaotong University;

【通讯作者】 郑向红;

【机构】 西安交通大学第二附属医院核医学科西安交通大学第二附属医院消化内科

【摘要】 背景:随着幽门螺杆菌(Hp)对抗菌药物耐药性和感染复发率的上升,高效清除Hp的难度日益增大。目的:探讨过氧化氢(H2O2)建立的高氧环境对Hp的清除作用及其可能机制。方法:使用溶解氧测定仪测定体内外条件下的H2O2溶液氧含量。以H2O2处理Hp标准菌株和野生型菌株;建立蒙古沙鼠Hp感染模型并予H2O2干预。紫外分光光度计检测Hp生长情况;常量肉汤稀释法测定最低抑菌浓度(MIC)和最低杀菌浓度(MBC);Giemsa染色观察胃黏膜Hp定植情况;透射电子显微镜和正苯基萘胺法观察Hp细胞膜形态学及其渗透性变化。结果:水溶液和家兔胃液中添加H2O2后,溶液氧含量显著升高(P均<0.05)。Hp在含H2O2液体培养基中的生长水平随H2O2浓度升高而逐渐降低,直至完全不生长。蒙古沙鼠经H2O2溶液灌胃14 d,胃黏膜定植的Hp数量接近于零。H2O2可诱导Hp细胞膜损伤、破裂,胞质溢出,细胞膜渗透性显著增高(P<0.05)。结论:H2O2能有效提高水溶液中的氧含量,在体内外条件下均能对Hp发挥高效清除作用,其机制与菌体细胞膜损伤密切相关。

【Abstract】 Background: Eradication of Helicobacter pylori(Hp) effectively is becoming exceedingly challenging due to the increase in antibiotic resistance and recurrence of Hp infection. Aims: To explore the effect of an oxygen-enriched environment established by hydrogen peroxide(H2O2) on elimination of Hp and the potential mechanism in vitro and in vivo. Methods: Oxygen concentration in H2O2 solution was measured by a dissolved oxygen meter in vitro and in vivo. Standard and wild-type Hp strains treated by H2O2 were used in study in vitro, while Mongolian gerbils infected with Hp and gavaged with H2O2 solution were used in study in vivo. Ultraviolet spectrophotometer was used to detect the bacterial growth. The minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) were determined by broth dilution method. The colonization of Hp in gastric mucosa was assessed by using Giemsa staining. Transmission electron microscope and N-phenyl-1-naphthylamine were used to detect the morphology and permeability of bacterial cell membrane. Results: Oxygen concentrations increased significantly after adding H2O2 to the aqueous solution and rabbit gastric juice(all P<0.05). Growth of Hp in the liquid medium containing H2O2 declined with rising H2O2 contents until it stopped growing completely. In Mongolian gerbils gavaged with H2O2 solution for 14 days, number of Hp colonized in gastric mucosa was close to 0 per high power field. While treated with H2O2, cell membrane of Hp was damaged and ruptured, cytoplasm overflowed, and the membrane permeability increased significantly(P<0.05). Conclusions: H2O2 increases oxygen concentration in liquid and eradicates Hp effectively in vitro and in vivo. Damage to bacterial cell membrane might be one of the potential mechanisms.

【基金】 国家自然科学基金青年科学基金项目(81700474);陕西省自然科学基础研究计划青年项目(2022JQ-838)
  • 【文献出处】 胃肠病学 ,Chinese Journal of Gastroenterology , 编辑部邮箱 ,2021年07期
  • 【分类号】R573
  • 【下载频次】28
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