食品与发酵工业 >

2022 , Vol. 48 >Issue 3: 85 - 91

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027894

研究报告

沙棘提取物对幽门螺杆菌脲酶活性的抑制

  • 刘欣 ,
  • 常应九 ,
  • 王树林 ,
  • 高庆超
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  • 1(青海大学 农牧学院,青海 西宁,810016)
    2(青海大学 省部共建三江源生态与高原农牧业国家重点实验室,青海 西宁,810016)
    3(江苏省食品质量安全重点实验室(江苏省农业科学院农产品质量与营养研究所),江苏 南京,210014)
本科生(王树林教授为通信作者,E-mail:wangsl1970@163.com)

收稿日期: 2021-04-28

  修回日期: 2021-06-19

  网络出版日期: 2022-03-04

基金资助

国家自然科学基金项目(32060543)

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Inhibition of Hippophae rhamnoides extracts on urease activity of Helicobacter pylori

  • LIU Xin ,
  • CHANG Yingjiu ,
  • WANG Shulin ,
  • GAO Qingchao
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  • 1(College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)
    2(State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining 810016,China)
    3(Key Laboratory of Food Quality and Safety of Jiangsu Province,Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Science,Nanjing 210014,China)

Received date: 2021-04-28

  Revised date: 2021-06-19

  Online published: 2022-03-04

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摘要

探讨沙棘提取物对幽门螺杆菌脲酶(Helicobacter pylori urease,HPU)活性的抑制及相关基因的表达影响。培养幽门螺杆菌(Helicobacter pylori)提取HPU,通过设置沙棘提取物系列浓度梯度及孵育时间梯度,采用Berthelot法检测HPU活性并评价沙棘提取物对HPU的抑制作用;进行酶活性动力学研究及对抑制位点进行分析;将菌液与不同浓度的沙棘提取物混合后孵育6 h,用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)法检测菌体中脲酶蛋白α亚基(urease subunit α,Ure-α),脲酶蛋白β亚基(urease subunit β,Ure-β),脲酶辅助蛋白E(urease accessory protein E,Ure-E)和脲酶辅助蛋白H(urease accessory protein H,Ure-H)脲酶基因的表达。结果表明,沙棘提取物对HPU的抑制作用呈时间、剂量依赖性,IC50值为(2.73±0.10)mg/mL。沙棘提取物对HPU的抑制类型为非竞争性抑制。巯基保护剂和Ni2+保护剂对HPU均无保护作用,而有协同沙棘提取物降低HPU活性的作用。沙棘提取物可降低HPU相关基因ure-α,ure-β,ure-E和ure-H的表达,对HPU具有良好的抑制效果。

关键词： 沙棘; 幽门螺杆菌; 脲酶; 基因表达

本文引用格式

刘欣 , 常应九 , 王树林 , 高庆超 . 沙棘提取物对幽门螺杆菌脲酶活性的抑制[J]. 食品与发酵工业, 2022 , 48(3) : 85 -91 . DOI: 10.13995/j.cnki.11-1802/ts.027894

Abstract

In order to investigate the effect of Hippophae rhamnoides Linn. extracts on inhibiting activity of Helicobacter pylori urease (HPU) and expression levels of related genes, the HPU was extracted and a series gradient of concentration for H. rhamnoides extracts and incubation time were set to evaluate inhibitory effect of the extracts on HPU by Berthelot method, and kinetic and inhibitory sites were analyzed. H. pylori was incubated with different concentration of H. rhamnoides extracts for 6 h, and RT-qPCR method was used to test the expression of ure-α, ure-β, ure-E and ure-H genes. The results showed that HPU was inhibited by H. rhamnoides extracts by time-dependent and dose-dependent manner, and IC50 values was (2.73±0.10) mg/mL. HPU was inhibited by H. rhamnoides extracts with noncompetitive type. Neither thiol nor Ni2+ protectors had effect on increasing the activity of HPU, contrarily, both of them coordinated with H. rhamnoides extracts to decrease the urease activity. The expression levels of ure-α, ure-β, ure-E and ure-H were decreased by extracts of H. rhamnoides extracts have a good inhibitory effect on HPU.

Key words： Hippophae rhamnoides Linn.; Helicobacter pylori; urease; gene expression

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