3种苯二氮[艹卓]氮类药物对幽门螺杆菌的抗菌作用及其机制

Inhibitory Effects and Mechanisms of Three Benzodiazepines on Helicobacter pylori

目的探究苯二氮[艹卓]氮类药物对幽门螺杆菌(Hp)的抗菌作用及其机制分析。方法以Hp国际标准菌株ATCC43504作为实验菌株,苯二氮[艹卓]类药物地西泮、咪达唑仑、瑞马唑仑为实验药物,阿莫西林、克拉霉素为阳性对照,注射用水为阴性对照,采用药敏纸片法测量每种药物的抑菌圈。采用抗菌效能检测药物对Hp的最低抑菌浓度(MIC)及最低杀菌浓度(MBC)。配置Hp菌悬液,分别用地西泮(地西泮组)、咪达唑仑(咪达唑仑组)处理,不加药的菌悬液作为对照组,使用全自动生化分析仪检测各组药物干预前(T_(0))及干预后1 (T_(1))、2 (T_(2))、3 (T_(3))、4 (T_(4))、5 (T_(5))、6 (T_(6))、7 h (T_(7))菌悬液中K^(+)浓度。提取Hp脲酶,分别采用1/2 MIC地西泮、MIC地西泮、2 MIC地西泮、1/2 MIC咪达唑仑、MIC咪达唑仑、2 MIC咪达唑仑、1 mg/ml乙酰氧肟酸、注射用水处理,酚红显色法检测各组pH值从6.8升至7.7所需的时间。结果地西泮、咪达唑仑、瑞马唑仑、阿莫西林、克拉霉素、注射用水对Hp的抑菌圈分别为52.3、42.7、6.0、72.3、60.8、6.0 mm。地西泮、咪达唑仑对Hp的MIC分别为12.5、25.0μg/ml,MBC分别为25、50μg/ml。与T_(0)比较,T_(1)~T_(7)地西泮组、咪达唑仑组和对照组的K^(+)浓度均明显升高(P均<0.01);T_(1)~T_(4)地西泮组和咪达唑仑组的K^(+)浓度明显高于对照组(P均<0.01)。1/2 MIC地西泮组、MIC地西泮组、2 MIC地西泮组、1/2 MIC咪达唑仑组、MIC咪达唑仑组、2 MIC咪达唑仑组对脲酶活性的抑制时间分别为(39.86±5.11)、(36.52±6.65)、(38.58±4.83)、(39.25±6.19)、(36.36±4.61)、(35.81±6.18) min,明显短于乙酰氧肟酸组(P均<0.01),但与注射用水组差异无统计学意义(P均>0.05)。结论苯二氮[艹卓]类药物地西泮、咪达唑仑对Hp有良好的抗菌作用,其抗菌机制可能与Hp细胞裂解有关,与脲酶抑制无关。

Objective To explore the inhibitory effects and mechanisms of benzodiazepines on Helicobacter pylori(Hp).Methods The Hp international standard strain ATCC43504 was treated with benzodiazepines diazepam,midazolam,and remimazolam,respectively.The treatments with amoxicillin and clarithromycin were taken as the positive controls,and that with water for injection as the negative control.The inhibition zone of each drug was measured by the disk diffusion method.The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of each drug against Hp were determined.Hp suspension was configured and treated with diazepam and midazolam,respectively.The bacterial suspension without drug added was used as the control group.The concentration of K^(+)in each bacterial suspension was measured by an automatic biochemical analyzer before drug intervention(T_(0))and 1(T_(1)),2(T_(2)),3(T_(3)),4(T_(4)),5(T_(5)),6(T_(6)),and 7 h(T_(7))after intervention.Hp urease was extracted and treated with 1/2 MIC diazepam,1 MIC diazepam,2 MIC diazepam,1/2 MIC midazolam,1 MIC midazolam,2 MIC midazolam,1 mg/ml acetohydroxamic acid,and water for injection,respectively.The time required for the rise from pH 6.8 to pH 7.7 in each group was determined by the phenol red coloring method.Results The inhibition zones of diazepam,midazolam,remimazolam,amoxicillin,clarithromycin,and water for injection against Hp were 52.3,42.7,6.0,72.3,60.8,and 6.0 mm,respectively.Diazepam and midazolam showed the MIC of 12.5μg/ml and 25.0μg/ml and the MBC of 25μg/ml and 50μg/ml,respectively,to Hp.The concentrations of K^(+)in the diazepam,midazolam,and control groups increased during T_(1)-T_(7) compared with those at T_(0)(all P<0.01).The concentration of K^(+)in diazepam and midazolam groups during T_(1)-T_(4) was higher than that in the control group(all P<0.01).The time of inhibiting urease activity in the 1/2 MIC diazepam,1 MIC diazepam,2 MIC diazepam,1/2 MIC midazolam,1 MIC midazolam,and 2 MIC midazolam groups was(39.86±5.11),(36.52±6.65),(38.58±4.83),(39.25±6.19),(36.36±4.61),and(35.81±6.18)min,respectively,which were shorter than that in the acetohydroxamic acid group(all P<0.01)and had no significance differences from that in the water for injection group(all P>0.05).Conclusion Diazepam and midazolam exerted inhibitory effects on Hp,which may be related to the cleavage of Hp cells rather than inhibiting urease.