酸枣果提取物在体内外对抗生素的增效作用及机制
投稿时间:2021-07-12  修订日期:2021-09-09  点此下载全文
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作者单位邮编
林金水 陕西省红枣重点实验室(延安大学)
延安大学生命科学学院 
716000
张 恒 陕西省红枣重点实验室(延安大学)
延安大学生命科学学院 
716000
高倩倩 陕西省红枣重点实验室(延安大学)
延安大学生命科学学院 
716000
王 凤 延安大学生命科学学院 716000
裴永福 延安大学生命科学学院 716000
成娟丽 陕西省红枣重点实验室(延安大学)
延安大学生命科学学院 
716000
张向前 陕西省红枣重点实验室(延安大学)
延安大学生命科学学院 
716000
基金项目:国家自然科学基金(32070103,31860012,31700031),陕西省教育厅重点实验室科研计划项目(17JS138);陕西省自然科学基础研究计划项目(2021JM-415),陕西省“特支计划”区域发展人才项目(陕组通字[2020]44号),陕西省普通高等学校青年杰出人才支持计划项目(陕高教组[2018]111号),延安大学博士科研启动项目(YDBK2016-01),国家级大学生创新创业训练项目(S202010719039),陕西省大学生创新创业训练项目(S202010719115)。
中文摘要:我们先前的研究显示,酸枣果的氯仿提取物具有广谱的抗菌增效作用,并从该氯仿提取物中进一步精制得到其低极性范围的活性组合物Fr.2a,发现Fr.2a与多种抗生素联用显示出广泛的协同抗菌作用。然而,由于Fr.2a的活性不及氯仿提取物,因此我们推测酸枣果氯仿提取物中其他极性范围内的物质可能也具有抗菌增效活性。为此本研究进一步对该酸枣果氯仿提取物中其他极性范围的抗菌増效活性物质进行分析和研究。在Fr.2a的基础上,本研究利用硅胶柱层析对酸枣果氯仿提取物中其他极性范围内的活性成分进行了分离纯化,得到精制物Fr.B。对精制物Fr.B进行GC-MS、核磁共振氢谱、红外光谱分析,抗菌增效谱的确定和抗菌增效机制的研究,并将精制物Fr.B制备成软膏,从体内和体外评价该软膏对抗生素的增效效果。结果表明:(1)由酸枣果氯仿提取物进一步精制得到的Fr.B组分,经过GC-MS、核磁共振氢谱、红外光谱分析显示它主要包含反油酸、油酸、顺-10-十六碳烯醇、棕榈酸等脂肪酸类化合物。(2)Fr.B与多种抗生素联用时显示出广泛的协同抗菌作用。(3)Fr.B的抗菌增效机制可能是通过改变细胞的通透性从而增强抗生素对耐药菌的抗菌作用。(4)在体内和体外Fr.B均能显著增强红霉素对耐甲氧西林金黄色葡萄球菌(MRSA)的杀菌作用,从而提高红霉素对MRSA菌株引起的伤口感染的治疗效果。综上,本研究所得到的Fr.B具有广谱的抗菌增效活性,能增强红霉素对伤口耐药菌感染的治疗效果。这为克服微生物对抗生素的耐药性提供了新的思路和解决方案。
中文关键词:酸枣果  精制物  脂肪酸  抗生素  抗菌增效剂
 
Synergistic Effect and Mechanism of Organic Extracts from Ziziphus jujuba Mill. var. spinosa Fruits on Antibiotics in Vitro and in Vivo
Abstract:Our previous studies have shown that the chloroform extracts from Ziziphus jujuba Mill. var. spinosa fruits has a broad-spectrum antimicrobial synergism. And we further extracted the low-polarity active ingredient Fr.2a from the chloroform extracts, and found that Fr.2a showed a wide range of synergistic antimicrobial effects when used in combination with a variety of antibiotics. However, since the activity of Fr.2a is inferior to that of chloroform extracts, we speculate that there were substances in the other polar range of chloroform extracts from Ziziphus jujuba Mill. var. spinosa fruits that might also have antimicrobial synergistic activities. For this reason, this study further analyzed and studied the antimicrobial synergists in other polar range of the chloroform extracts from Ziziphus jujuba Mill. var. spinosa fruits. On the basis of Fr.2a, the active components in other polar range of the chloroform extracts from Ziziphus jujuba Mill. var. spinosa fruits were separated and purified by silica gel column chromatography, and the refined product Fr.B was obtained. Then, we analyzed Fr.B by GC-MS, hydrogen nuclear magnetic resonance spectroscopy (1H-NMR), infrared spectroscopy (IR), determination of antimicrobial synergistic spectrum and study of antimicrobial synergistic mechanism, and prepared Fr.B into ointment, and evaluated the synergistic effect of the ointment on antibiotics in vivo and in vitro. The results were as follows: (1) The Fr.B component obtained by further refining the chloroform extracts of Ziziphus jujuba Mill. var. spinosa fruits, after GC-MS, 1H-NMR, and IR analysis showed that it mainly contains fatty acid compounds such as elaidic acid, oleic acid, cis-10-hexadecenol, palmitic acid and so on. (2) Fr.B showed a wide range of synergistic antimicrobial effects when combined with a variety of antibiotics. (3) The antimicrobial synergistic mechanism of Fr.B may be to enhance the antimicrobial effect of antibiotics on resistant organism by changing the permeability of cells. (4) Fr.B could significantly enhance the bactericidal effect of erythromycin on methicillin-resistant Staphylococcus aureus (MRSA) in vivo and in vitro, thereby improving the therapeutic effect of erythromycin on wound infections caused by MRSA strains. In conclusion, Fr.B obtained in this study has a broad spectrum of antimicrobial synergistic activity, which can enhance the efficacy of erythromycin in the treatment of wound drug-resistant bacteria infection. This provides new ideas and solutions for overcoming microbial resistance to antibiotics.
keywords:Ziziphus jujuba Mill. var. spinosa fruit  refined extract  fatty acid  antibiotics  antimicrobial synergist
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