黄连素对幽门螺杆菌多重耐药株hefABC基因表达的影响  

The Effects of Berberine in the Expression Quantities of Efflux Pump Genes hef'ABC of Helicobacter Pylori of Multi-drug resistance

作　　者：周帆 徐小青[2] 彭君伟 曹勤[2] Zhou Fan;Xu Xiaoqing;Peng Junwei;Cao Qin(Gastroenterology Department,The Third People’s Hospital of Huzhou,Huzhou,Zhejiang 313000,China;Gastroenterology Department,Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Department of integrated Chinese and Western medicine,Shanghai Minhang District Tumor Hospital,Shanghai 220240,China)

机构地区：[1]湖州市第三人民医院消化内科,浙江湖州313000 [2]上海中医药大学附属普陀医院消化内科,上海200062 [3]上海市闵行区肿瘤医院中西医结合科,上海220240

出　　处：《哈尔滨医药》2024年第4期1-4,共4页Harbin Medical Journal

基　　金：国家自然科学基金(青年)项目(81603468);湖州市科技局公益性应用研究项目(2023GYB15)。

摘　　要：目的研究黄连素(BBR)在幽门螺杆菌(Hp)多重耐药(MDR)菌株外排泵基因hefABC调控中的作用。方法①通过E-test药敏实验检测阿莫西林(AMX)、左氧氟沙星(LEV)、克拉霉素(CLR)、甲硝唑(MTZ)对临床分离Hp菌株的最小抑菌浓度(MIC),从中筛选出敏感株和MDR株;②采用实时荧光定量聚合酶链反应(RT-PCR)测定敏感株与MDR株中外排泵基因hefABC mRNA的表达水平;③测定黄连素对MDR株的MIC值,在1/2 MIC黄连素浓度下液体培养MDR菌株,培养5代后用RT-PCR测定黄连素作用前后hefABC mRNA的表达量变化。结果①从60株分离菌株中筛选出敏感株15株,MDR株12株;②与敏感组相比,外排泵基因hef'A、hefB的表达量在MDR组中明显升高,差异有统计学意义(P<0.05),hefC基因的表达水平在两组之间无明显差异;③黄连素作用后外排泵基因hef'A、hefB的表达量与作用前相比明显降低,差异有统计学意义(P<0.05)。结论①Hp的多重耐药与外排泵基因hefA、hefB表达量的上调有关。②黄连素通过下调外排泵基因hefA、hefB的表达抑制部分MDR菌株的耐药性。Objective To study the the role of berberine(BBR)in the regulation of efflux pump genes hefABC in multi-drug resistant(MDR)strains of Helicobacter pylori(Hp).Methods①Hp clinical isolates were tested the minimum inhibitory concentration for amoxicillin,levofloxacin,clarithromycin and metronidazole by E-test method.,in which sensitive strains and MDR strains were screened.②The expression quantities diffenrence of efflux pump genes hefABC between MDR group and sensitive group were detected by Real-time fluorescent quantitative polymerase chain reaction(RT-PCR).③The MIC values of berberine against MDR strains were determined by agar two-fold dilution method.After the MDR strains were cultured at 1/2 MIC concentration of berberine in liquid for 5 generations,RT-PCR was used to detect the mRNA expression difference of the efflux pump genes before and after the addition of berberine.Results①A total of sixty Hp strains were isolated and screened including fifteen sensitive strains and twelve MDR strains.②The expression quantities of hefA、hefB in the MDR group are significantly higher than that in the sensitive group(P<0.05).There are no high expressions of hefC between two groups(P<0.05).③The expression quantities of Hp0605,Hp0606 of MDR group after the effects of berberine remarkably decrease,compared with that before treated with berberine,and the difference has statistical significance.Conclusions①Multi-drug resistance of H.pylori is related to the high expression of efflux pump genes of Hp0605,Hp0606.②Berberine can inhibit drug resistance in some MDR strains by down-regulating the expression of efflux pump genes hefA and hefB.

关 键 词：黄连素 幽门螺杆菌 多重耐药 外排泵基因 

分 类 号：R573.6[医药卫生—消化系统]