溶菌酶与EDTA二钠的协同抑菌作用

摘要/Abstract

摘要： 目的：研究溶菌酶与乙二胺四乙酸二钠（ethylenediaminetetraacetic acid disodium salt，EDTA二钠）对粪肠球菌和牙髓卟啉单胞菌的协同抑菌作用。方法：分别培养粪肠球菌和牙髓卟啉单胞菌，并调整菌液浓度至108菌落形成单位（colony-forming unit，CFU）/mL；配制终浓度为0.3、0.5、1、2、5、10、50、100、150和300 g/L的单纯溶菌酶抑菌液，以及添加浓度为0.5、1.0、2.0 g/L的EDTA二钠的混合抑菌液。将菌液与抑菌液作用15 min，加入水溶性四唑盐（water-soluble tetrazolium，WST）工作液染色，酶标仪测定光密度值，计算细菌活性。结果：单纯溶菌酶抑菌液（浓度0.5~150 g/L）对两种细菌的抑菌作用随溶菌酶浓度的升高而增强，且对粪肠球菌的抑菌作用更显著。EDTA二钠与溶菌酶有协同抑菌作用，与溶菌酶浓度相关。溶菌酶浓度为0.5~50 g/L时，EDTA二钠协同溶菌酶作用于粪肠球菌（P<0.05），可使其抑菌作用提高约1.2~3.7倍；溶菌酶浓度为0.5~10 g/L时， EDTA二钠协同溶菌酶作用于牙髓卟啉单胞菌（P<0.05），可使其抑菌作用提高约1.3~3.5倍；当溶菌酶浓度大于100 g/L时，加入EDTA二钠对细菌均没有明显的协同抑菌作用（P>0.05）。结论：对于粪肠球菌及牙髓卟啉单胞菌，在溶菌酶较低浓度时，EDTA二钠有协同抑菌作用；在溶菌酶较高浓度时，EDTA二钠无协同抑菌作用。

关键词: 胞壁质酶, 依地酸, 药物协同作用, 抗菌药

Abstract: Objective: To evaluate the synergistic antibacterial effects of lysozyme with ethylenediaminetetraacetic acid disodium salt (EDTA-2Na) on Enterococcus faecalis (E. faecalis) and Porphyromonas endodontalis (P. endodontalis). Methods: E. faecalis and P. endodontalis were cultured and adjusted to 10⁸CFU/mL. Then 0.3, 0.5, 1, 2, 5, 10, 50, 100, 150 and 300 g/L of lysozyme were prepared with deionized water; and the lysozyme solutions were mixed with 0.5, 1.0, 2.0 g/L of EDTA-2Na, respectively. The bacteria and lysosome with/without EDTA-2Na interacted for 15 min, then water-soluble tetrazolium (WST) working solution was added and the activity of the bacteria was calculated by measuring optical densities at 450 nm and 630 nm with microplate spectrophotometer. Results: Regarding the pure lysozyme from 0.5 g/L to 150 g/L, more E. faecalis and P. endodontalis were inhibited when the concentration of lysozyme was higher, especially for E. faecalis. There was synergistic effect of lysozyme with EDTA-2Na on antibacterial activity, which was related to the concentration of lysozyme. On E. faecalis, the antibacterial activity of lysozyme with EDTA-2Na was 1.2-3.7 folds than the pure lysozyme when the concentration of lysozyme was 0.5-50 g/L (P<0.05), and on P. endodontalis, the antibacterial activity of lysozyme with EDTA-2Na was 1.3-3.5 folds than the pure lysozyme when the concentration of lysozyme was 0.5-10 g/L (P<0.05). When the concentration of lysozyme was higher than 100 g/L, EDTA-2Na did not show synergistic effect on the antibacterial activity (P>0.05). Conclusion: For E. faecalis and P. endodontalis, a low concentration of lysozyme with EDTA-2Na showed significant synergistic antibacterial activity, while a high concentration of lysozyme with EDTA-2Na did not.

Key words: Muramidase, Edetic acid, Drug synergism, Anti-bacterial agents

李小曼, 王晓燕, 高学军. 溶菌酶与EDTA二钠的协同抑菌作用[J]. 北京大学学报（医学版）, 2015, 47(1): 52-56.

LI Xiao-Man, WANG Xiao-Yan, GAO Xue-Jun. Synergistic effects of lysozyme with EDTA-2Na on antibacterial activity[J]. Journal of Peking University(Health Sciences), 2015, 47(1): 52-56.

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