Antibacterial effect of emodin on multi-resistant staphylococcus aureus and its mechanism
ZHU Kunpeng1, HUANG Shiqi2, HAN Yizhen3
1. Procurement Service Department, 3. Dental Department, Characteristics Medical Center of Chinese People’s Armed Police Force, Tianjin 300162, China; 2. Outpatient Department of Nankai University,Tianjin 300071, China
Abstract:Objective To explore antibacterial effect of emodin on multi-resistant staphylococcus aureus (staphylococcus multidrug-resistant) and its mechanism. Methods The drug resistance of clinically isolated methicillin-resistant staphylococcus aureus (MRSA) was determined by full-automatic microbial identification and drug sensitivity analyzer. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and the half inhibitory concentration (IC50) of emodin in MRSA standard strains and clinically isolated strains were detected by double dilution method. With MRSA standard strains as the research background, they were divided into control group, emodin (1×MIC) group and emodin (2×MIC) group. The number of bacteria in MRSA standard strains was observed during a period of time (0, 0.5, 1, 2, 4, 8, 16 h). The permeability of cell membrane was observed by double-color fluorescent staining. The levels of biofilm-related genes were detected by real-time quantitative PCR (RT-PCR). The models of mice with MRSA infection were prepared, and they were divided into infection group, emodin (2.5 mg/kg) group, emodin (5 mg/kg) group and emodin (10 mg/kg) group. The mice models without infection treatment were enrolled as control group. At 1h after infection, mice in all emodin groups were given corresponding doses of emodin. At 12h after administration, bacteria content in blood, inflammatory factors and phagocytic function of peritoneal macrophages were compared. Results The results of drug-resistant bacteria confirmed that the 4 clinically isolated strains were MRSA. MIC, MBC and IC50 of emodin for standard and clinically isolated MRSA strains were (4, 8 , 8, 8, 2 μg/ml),(8, 16, 8, 8, 4 μg/ml) and (2.51, 4.15, 5.36, 4.01, 1.43 μg/ml), respectively. The number of standard MRSA strains, expressions of cidA, icaA, agrA, sortaseA and sarA in emodin (1×MIC) group and emodin (2×MIC) group were lower than those in control group (P<0.05), and there was red fluorescence in fluorescence images. The levels of drug-resistant bacteria in blood, serum tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), interleukin-8 (IL-8), interferon gamma (INF-γ) and procalcitonin (PCT) in emodin (2.5 mg/kg) group, emodin (5 mg/kg) group and emodin (10 mg/kg) group were lower than those in infection group (P<0.05), phagocytic rate and index of peritoneal macrophages were higher than those in infection group (P<0.05). Conclusions Emodin can improve the permeability of cell membrane, regulate the expression of biofilm-related genes, reduce inflammation level and recover the phagocytic function of macrophages.
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