小檗碱联合阿奇霉素对小鼠腹腔金黄色葡萄球菌早期生物被膜感染的体内影响

In vivo effects of berberine combined with azithromycin on early biofilm infection of Staphylococcus aureus in mice

目的:探讨小檗碱(berberine,Ber)联合阿奇霉素(azithromycin,Azm)对小鼠金黄色葡萄球菌(staphylococcus aureus,SA)被膜早期(biofilm,BF)感染的治疗作用。方法:以输液管为载体,金黄色葡萄球菌(ATCC25923)为实验菌株,体外培养形成早期金黄色葡萄球菌生物被膜,建立小鼠腹腔生物被膜感染模型。建模后立即给予药物干预,分为Ber组(30 mg/kg)灌胃、AZM组(30 mg/kg)腹腔注射、Ber(15 mg/kg)灌胃+AZM(15 mg/kg)组腹腔注射、空白组给予生理盐水腹腔注射,给药24 h后取出载体,根据连续稀释法对载体表面进行活菌计数,扫描电子显微镜(SEM)观察载体表面BF形态特征变化,载体周围组织HE染色进行组织病理学分析。结果:在药物作用24 h后,对载体进行表面菌落计数,单独给药组及联合给药组菌落数都少于空白对照组(P<0.01);Ber+AZM组菌落数较Ber、AZM组单独给药组显著变少(P<0.01);空白对照组SEM观察BF生长形态均比Ber、AZM、Ber+AZM组形成的BF稠密且繁多;AZM组较Ber组稀疏;Ber+AZM组BF基本被清除,只见少量金黄色葡萄球菌黏附。腹腔感染组织病理学表现空白对照组炎症最为明显,Ber+AZM组只见轻度的炎症反应。结论:Ber对小鼠腹腔SA感染早期BF的形成具有干扰作用,其与AZM联合给药明显增强对小鼠腹腔SA早期BF感染的治疗作用。

Objective:To test the efficacy of Berberine combined with Azithromycin treatment of early biofillm infection with Staphylococcus aureus(S.aureus)in intraperitoneal cavities of mice.Methods:The infusion tube was used as carrier,the S.aureus strains(ATCC25923)were set as the experimental strains.The early biofilm of S.aureus was formed by in intro culture,After the model of intraperitoneal implant BF infection was established,the mice in the Ber-treated group(30mg/kg)were treated with intragastric administration of Ber,the mice in the AZM-treated group(30mg/kg)were treated with intraperitoneal injection of AZM,Ber(15mg/kg)intragastric administration+AZM(15mg/kg)group intraperitoneal injection,blank group intraperitoneal injection of normal saline.The implants were removed out from the peritoneal cavities after the drug treatment for 24 hours,the bacterial colony counting was conducted by using continuous dilution method,the morphological changes of BF on the surfaces of implants were observed under scanning electron microscope(SEM),and the histopathological changes of tissue sections of abdominal infection were observed under HE staining microscope.Results:The bacterial colony counts on surfaces of implants were significantly less in the drug groups than that in the control group after the drug treatment for 24 hoursb(P<0.01).The bacterial colony counts were significantly less in the Ber+AZM group than in the Ber group and the AZM group(P<0.01).The SEM observation showed that the BF formed on the surfaces of implants was much and thick in the control group and the drug groups,the BF of the Ber+AZM was remarkably thin,only a few S.aureus adhesions were seen.As for the histopathological manifestations of abdominal infection,the inflammatory manifestations of the control group were the most obvious;only mild inflammatory reactions were seen in the Ber+AZM group.Conclusion:The single use of Ber may inhibit the growth of BF in intraperitoneal cavities of mice with early S.aureus infection,and its combination with AZM can remarkably enhance the bactericidal effect of BF.