摘要
目的 研究人参皂苷Rh2联合万古霉素对万古霉素耐药型耐甲氧西林金黄色葡萄球菌生物膜和抑菌效果的影响。方法 诱导万古霉素耐药型耐甲氧西林金黄色葡萄球菌(VRSA),分别用10μg/m L、30μg/mL、90μg/m L人参皂苷Rh2处理VRSA作为不同浓度人参皂苷Rh2处理组;VRSA分别用1μg/mL、4μg/mL、16μg/mL万古霉素处理,作为不同浓度万古霉素处理组;10μg/mL人参皂苷Rh2和1μg/mL万古霉素共同作用VRSA记为10μg/mL人参皂苷Rh2+1μg/mL万古霉素组;不作处理的VRSA作为空白对照组。原始分离的对万古霉素敏感型的耐甲氧西林金黄色葡萄球菌作为VSSA组。结晶紫法检测生物膜形成能力;实时荧光定量PCR (RT-qPCR)检测fnbA、atlA表达水平;分光光度计检测菌活性。结果 与VSSA组比较,VRSA组生物膜的光密度值及fnbA、atlA的表达水平均明显升高,差异有统计学意义(P<0.05);与空白对照组比较,16μg/mL万古霉素及10μg/mL人参皂苷Rh2+1μg/mL万古霉素处理的VRSA增殖活性明显降低,而10μg/mL人参皂苷Rh2+1μg/mL万古霉素处理的VRSA增殖活性明显低于不同浓度人参皂苷Rh2处理组,且明显高于4μg/mL万古霉素处理者,差异均有统计学意义(P<0.05);与空白对照组比较,不同浓度人参皂苷Rh2处理的VRSA生物膜的光密度值明显降低,fnbA、atlA的表达水平明显降低,差异均有统计学意义(P<0.05);与空白对照组比较,不同浓度万古霉素处理的VRSA中fnbA、atlA表达水平显著升高,4μg/mL和16μg/mL万古霉素处理的VRSA生物膜的光密度值也明显升高,差异均有统计学意义(P<0.05);与空白对照组比较,10μg/mL人参皂苷Rh2组及10μg/mL人参皂苷Rh2+1μg/mL万古霉素组VRSA生物膜的光密度值及fnbA、atlA的表达水平明显降低,fnbA、atlA的表达水平明显升高,且10μg/mL人参皂苷Rh2+1μg/mL万古霉素组VRSA生物膜的光密度值及fnbA、atlA的表达水平均显著高于10μg/mL人参皂苷Rh2组,但明显低于1μg/mL万古霉素组,差异均有统计学意义(P<0.05)。结论 人参皂苷Rh2可抑制VRSA生物膜形成,但不抑菌;万古霉素可抑菌但不能抑制生物膜形成,两者联用能抑制生物膜形成且增强万古霉素的抑菌作用。
Objective To investigate the effects of ginsenosides Rh2 combined with vancomycin on vancomycin-resistant methicillin-resistant Staphylococcus aureus biofilms and antibacterial effects. Methods Vancomycin-resistant methicillin-resistant Staphylococcus aureus(VRSA) was induced and treated with 10 μg/mL, 30 μg/mL, 90 μg/mL ginsenosides Rh2 as different concentrations of ginsenoside treatment groups. VRSA was treated with 1 μg/mL, 4 μg/mL,16 μg/mL vancomycin as different vancomycin-treated groups. VRSA was treated with 10 μg/mL ginsenoside Rh2 and1 μg/mL vancomycin as 10 μg/mL ginsenoside Rh2+1 μg/mL vancomycin group. VRSA without treatment was used as a blank control group. The originally isolated vancomycin-sensitive Staphylococcus aureus(VSSA) were enrolled as VSSA group. The crystal violet method was used to detect the biofilm formation ability. Real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expressions of fnbA and atlA level. The spectrophotometer was used to detect the bacterial activity. Results Compared with VSSA group, the optical density value of biofilm and the expression levels of fnb A and atlA in VRSA group were significantly increased(P<0.05). Compared with the blank control group, the proliferative activity of VRSA treated with 16 μg/mL vancomycin and 10 μg/mL ginsenoside Rh2+1 μg/mL vancomycin decreased significantly;and the proliferation activity of VRSA treated with 10 μg/mL ginsenoside Rh2+1 μg/mL vancomycin was significantly lower than that treated with different concentrations of ginsenoside Rh2 and significantly higher than that treated with 4 μg/mL vancomycin, with statistically significant differences(P<0.05). Compared with the blank control group, the optical density of VRSA biofilm treated with different concentrations of ginsenoside Rh2 decreased significantly, and the expression levels of fnbA and atlA decreased significantly(P<0.05). Compared with the blank control group, the expression levels of fnbA and atlA in VRSA treated with different concentrations of vancomycin were significantly increased, and the optical density of VRSA biofilm treated with 4 μg/mL and 16 μg/mL vancomycin increased significantly(P<0.05). Compared with the blank control group, the optical density and the expression levels of fnbA and atlA of VRSA biofilm in 10 μg/mL ginsenoside Rh2 group and 10 μg/mL ginsenoside Rh2+1 μg/mL vancomycin group were significantly decreased, while the expression levels of fnbA and atlA were significantly increased. The optical density of VRSA biofilm and the expression levels of fnb A and atlA in 10 μg/mL ginsenoside Rh2+1 μg/mL vancomycin group were significantly higher than those in 10 μg/mL ginsenoside Rh2 group, but significantly lower than those in 1 μg/mL vancomycin group(P<0.05). Conclusion Ginsenosides Rh2 can inhibit the formation of VRSA biofilms, but can not inhibit bacteria. Vancomycin can inhibit bacteria but not biofilms, and the combination of them can inhibit biofilm formation and enhance the antibacterial effect of vancomycin.
作者
张霄霄
邵海连
戈伟
汪定成
董轲
应后群
ZHANG Xiao-xiao;SHAO Hai-lian;GE Wei;WANG Ding-cheng;DONG Ke;YING Hou-qun(Department of Laboratory Medicine,the Second Affiliated Hospital of PLA Air Force Military Medical University,Xi'an 710038,Shaanxi,CHINA;Department of Nuclear Medicine,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi,CHINA)
出处
《海南医学》
CAS
2023年第4期457-461,共5页
Hainan Medical Journal
基金
国家自然科学基金(编号:81702090)。
作者简介
第一作者:张霄霄(1988-),女,硕士,主管检验技师,主要研究方向为临床微生物检验工作及细菌耐药性研究;通讯作者:董轲(1971-),男,博士,副主任医师,主要研究方向为病毒或肿瘤疫苗、肿瘤分子生物学研究,E-mail:zhanjinper@163.com。
