新型抗菌活性氨基酸卟啉光敏剂4I体外研究  

In vitro study of the antibacterial activity of novel amino acid-porphyrin photosensitizer 4I

作　　者：孙实[1] 靳衡[1] 袁瑶[2] 丁辉[2] 洪阁[3] 刘子泉[2] 寿松涛[1] 侯世科[2] 

机构地区：[1]天津医科大学总医院急诊医学科,300052 [2]武警后勤学院附属医院,武警部队救援医学研究所,天津300162 [3]中国医学科学院 北京协和医学院生物医学工程研究所,天津300192

出　　处：《国际生物医学工程杂志》2015年第6期340-344,共5页International Journal of Biomedical Engineering

基　　金：中国人民武装警察部队后勤学院灾害应急救援医学全军重点实验室开放基金（JY1405）;天津市卫生局科技基金（2015KZ123）

摘　　要：目的评价新型氨基酸卟啉光敏剂4I对临床分离的敏感菌株和多重耐药菌株的抗菌活性；分别检测其对人永生化表皮细胞HaCaT及单核巨噬细胞RAW264．7的细胞毒性，为体内实验提供基础。方法用二倍稀释法将不同浓度的光敏剂与菌悬液避光孵育30min后，立即行红色弱激光照射30min（650nm，6J／cm^2），采用涂板计数法评定4I对不同菌株的最小抑菌浓度（MIC）和最小杀菌浓度（MBC），并计算MIC50和MIC90；CCK．8法分别检测4I对HaCaT及RAW264．7细胞的细胞毒性。结果除奇异变形杆菌和摩根摩根菌外，4I对其余菌株均有效；灭活同一菌种的敏感菌和耐药菌效果差异无统计学意义（P〉0．05）；MIC50及MIC90分别为0．822μmol／L和1．947μmol／L；单独光敏剂孵育及单纯红色弱激光照射30min后，显示有促HaCaT细胞生长作用（P〈0．05），对RAW264．7细胞存活率则无影响；当4I的浓度为0．98μmol／L时，HaCaT和RAW264．7细胞的存活率分别约为92．24％±13．64％和103．84％±1．59％，半抑制浓度（IC50）分别为1．846μmol／L和7．447μmol／L，表明在有效抑菌浓度下对细胞毒副作用较弱。结论新型氨基酸卟啉类光敏剂4I具有高效、广谱的抗菌活性，无细胞毒性，低毒副作用，且具有促细胞生长作用，是一种前景极佳的抗菌光敏剂，将其结合红色弱激光的光动力抗菌疗法具有广阔的应用前景。Objective To evaluate the antibacterial activity of novel amino acid-porphyrin photosensitizer against 11 kinds of clinical isolated sensitive and multi-resistance strains, and to test its cytotoxicity to HaCaT and RAW264.7 cells, in order to provide basis for in vivo experiment. Methods Two-fold dilution method was used to co-cuhure different concentration photosensitizers with bacterial suspension for 30 min in the dark, and the culture mix were immediately irradiated by red laser （650 nm, 6 J/cm^2） for 30 rain. After irradiation, the minimum inhibitory concentration （MIC） and minimum bactericidal concentration （MBC） values of 4I to different strains were evaluated using tablet coated plate method, the MIC50 and MIC90 values were calculated, and the cytotoxicity of 4I to HaCaT and RAW264.7 cells were determined by CCK-8 assay. Results The results showed that, except Proteus mirabilis and Morganella strains, 4I was efficient to all of the rest strains, and there was no variability of the effectiveness between sensitive and multi-resistance strains. The MICS0 and MIC90 values were 0.822 μmol/L and 1.947 μmol/L, respectively. Photosensitizer or irradiated by red laser alone showed mild promotion on HaCaT cells growth （P〈0.05）, while no effect was observed on RAW264.7 cells. After photodynamic therapy, survival rates of HaCaT and RAW264.7 cells under 4I concentration of 0.98 μmol/L were （92.24±13.64）% and （103.84±1.59）%, respectively. IC50 values were 1.846 μmol/L and 7.447 μmol/L, which suggested low-cytotoxicity effect under efficient inhibitory bacterial concentration. Conclusions The novel amino acid-porphyrin photosensitizer 4I has high efficiency and broad spectrum antimicrobial properties, no cytotoxicity, and low toxic side effect. It has promoting effect on cell growth, and is a promising antimicrobial photosensitizer. The antimicrobial therapy of photosensitizer with lower power red laser has more broad application prospect.

关 键 词：光敏药物 多重耐药菌株 细胞毒性 

分 类 号：R96[医药卫生—药理学]