摘要
目的:研究大黄酸体内外抗耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的活性及机制。方法:体外采用二倍稀释法确定大黄酸的最低抑菌浓度(MIC)和最低杀菌浓度(MBC),通过检测药物处理后MRSA凋亡情况、对细菌细胞膜和菌体总蛋白合成的影响,初步探讨大黄酸抗MRSA的相关作用机制;体内利用尾静脉注射MRSA菌液建立小鼠菌血症模型,记录检测小鼠精神状态、体质量、存活率、血液荷菌量、主要器官荷菌量及病理改变等指标来对大黄酸体内抗菌活性进行评估与分析。结果:大黄酸对MRSA的MIC和MBC分别为8μg/mL和16μg/mL;与空白对照组相比,大黄酸8、16μg/mL处理MRSA后,能够有效诱导细菌进入凋亡期(P<0.01),显著提高菌液的碱性磷酸酶含量和电导率(P<0.01),菌体总蛋白条带模糊或消失,且细菌超微结构受到破坏;与模型对照组小鼠相比,大黄酸2.5、5、10 mg/kg组的小鼠精神状态、体质量以及活跃度良好,存活率得到明显提高,体内血液及重要器官荷菌量显著降低(P<0.01),HE染色结果表明,大黄酸给药组小鼠内脏病变有明显的好转。结论:大黄酸主要通过破坏细菌的细胞膜,抑制细菌的生长和繁殖,从而在体内体外对MRSA发挥较强的抗菌作用。
Objective:To study the in vitro and in vivo activity of rhein against methicillin-resistant Staphylococcus aureus(MRSA)and the mechanism.Methods:For the in vitro activity,the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of rhein were determined with the two-fold serial dilution method.The apoptosis of MRSA,bacterial cell membrane,and total proteins in the cells were examined after the treatment with rhein to preliminarily discuss the mechanism of rhein against MRSA.As for the in vivo activity,MRSA solution was injected into the tail vein of mice to induce bacteremia,and the mental state,body weight,survival rate,bacterial load in blood,bacterial load in major organs,and pathological changes of mice were recorded.Results:The MIC and MBC of rhein against MRSA were 8μg/mL and 16μg/mL,respectively.Compared with the blank control group,8μg/mL and 16μg/mL rhein induced the apoptosis of MRSA(P<0.01)and improved the alkaline phosphatase(AKP)content and electrical conductivity of the bacterial fluid(P<0.01).In addition,bands of the total proteins in the bacteria blurred or disappeared and the bacterial ultrastructure was damaged.Compared with the model control group,mice in the 2.5,5,and 10 mg/kg rhein groups were outstanding in the mental state,body weight and activity,and the survival rate was significantly raised and the bacterial load in blood and vital organs was decreased significantly in the three rhein groups(P<0.01).Hematoxylin and eosin(HE)staining suggested the obvious alleviation of visceral lesions of mice treated with rhein.Conclusion:Rhein mainly destroys the membrane of MRSA to inhibit its growth and reproduction,thereby exerting the strong inhibitory effect on MRSA in vitro and in vivo.
作者
李洁
王琦
张群
刘洋
覃静
任瑶瑶
蒋合众
谭睿
Li Jie;Wang Qi;Zhang Qun;Liu Yang;Tan Jing;Ren Yaoyao;Jiang Hezhong;Tan Rui(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610030)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第3期70-76,共7页
Pharmacology and Clinics of Chinese Materia Medica
基金
四川省政府应急重大专项(编号:2021XYCZ008)
四川省药品监督管理局中药(民族药)标准提升(第二次)(编号:510201202102305)
四川省中医药强省发展中心(编号:510201202109711)
国家自然科学基金面上项目(编号:82074003)
