黄连水煎剂对金黄色葡萄球菌生物膜的作用及与苯唑西林钠联合作用的研究

来源期刊：广州医药 | 13-17 发布时间：2021-12-01 收稿时间：2025/11/13 17:24:41 阅读量：36

作者：: 李奕铮,

潘玲,

廖蕴华,

关键词：: 黄连水煎剂金黄色葡萄球菌生物膜苯唑西林钠; Coptis decoction Staphylococcus aureus Biofilm Oxacillin sodium

DOI：: 10.3969/j.issn.1000-8535.2018.01.004

收稿时间：: 2017-10-01
修订日期：
接收日期：
引用总数：: 0

目的研究黄连水煎剂对体外建立的金黄色葡萄球菌生物膜(bacterial biofilm,BF)的作用,及黄连水煎剂联合苯唑西林钠对金黄色葡萄球菌作用。方法由广西医科大学第一附属医院检验科提供金黄色葡萄球菌菌株34134,采用二倍稀释法分别测出黄连水煎剂和苯唑西林钠对金黄色葡萄球菌的最低抑菌浓度(minimum inhibitory concentration MIC),在体外使用腹膜透析管建立金黄色葡萄球菌生物膜模型,加入不同浓度的黄连水煎剂和苯唑西林钠作用3天和7天后,琼脂平板菌落计数法评估细菌粘附腹透管的能力,结晶紫染色法行载体表面BF半定量,银染法快速鉴定BF。结果黄连水煎剂MIC为16 mg/mL,苯唑西林钠MIC为4 μg/mL,3天BF模型中,1/2MIC黄连水煎剂、1/4MIC黄连水煎剂、1/8MIC黄连水煎剂、1/16MIC黄连水煎剂、1/2MIC苯唑西林钠、1/4MIC苯唑西林钠作用下的金黄色葡萄球菌的菌落计数分别为(39.333 3±3.994 2)×10⁶ CFU/mL、(52.366 7±3.537 8)×10⁶ CFU/mL、(81.266 7±3.341 8)×10⁶ CFU/mL、(90.900 0±2.040 1)×10⁶ CFU/mL、(50.866 7±2.208 6)×10⁶ CFU/mL、(77.666 7±2.880 7)×10⁶ CFU/mL,空白组的菌落计数为(92.033 3±3.890 6)×10⁶ CFU/mL,与空白组相比较,1/2MIC黄连水煎剂,1/4MIC黄连水煎剂、1/8MIC黄连水煎剂、1/2MIC苯唑西林钠、1/4MIC苯唑西林钠的差异有统计学意义(P<0.001),1/16MIC黄连水煎剂与空白组比较,差异无统计学意义(P=0.173)。7天的BF模型中,1/2黄连水煎剂比1/2苯唑西林钠抑制金黄色葡萄球菌生物膜形成的作用更强(P<0.001),结晶紫、银染也得到相似的结果,进一步的研究发现在1/16MIC黄连水煎剂的作用下,加入不同浓度的苯唑西林钠,此时苯唑西林钠的最低抑菌浓度为0.25 μg/mL。结论黄连水煎剂能抑制金黄色葡萄球菌生物膜的形成,并且在苯唑西林钠联合黄连水煎剂时,能有效地提高苯唑西林钠的疗效,为治疗金黄色葡萄球菌导致的腹膜透析相关性腹膜炎提供新的治疗方案,同时也提高了抗生素的疗效,为临床上减少耐药菌的产生提供了帮助。

Objective To study the effect of Coptis chinensis decoction on bacterial biofilm(BF)in vitro and the effect of Coptis chinensis decoction combined with oxacillin sodium on Staphylococcus aureus. Methods Staphylococcusaureus strain 34 134 was provided by the First Affiliated Hospital of Guangxi Medical University. Using doubling dilution method to measure the minimum inhibitory concentration of Coptis decoction and Oxacillin sodium on Staphylococcus aureus,respectively. In vitro, peritoneal dialysis tube was used to establish Staphylococcus aureus biofilm model after adding different concentrations of Coptis decoction and Oxacillin sodium for 3 days and 7 days, we used Agar plate count method to assess bacterial adhesion on the dialysis tube, crystal violet staining and semi quantitative method to measure the BF of the surface of the carrier and silver staining(light microscopy) rapid identification of BF. Results The MIC of Coptidis decoction was 16 mg/ml, and that of Oxacillin sodium was 4 μg/mL. In the BF model after 3 days, the colony counting of bacteria of 1/2MIC Coptis decoction, 1/4MIC Coptis decoction, 1/8MIC Coptis decoction,1/16MIC Coptis decoction, 1/2MIC Oxacillin sodium,1/4MIC Oxacillin sodium were(39.333 3+3.994 2)×10⁶(52.366 7+3.537 8),CFU/mL×10⁶, CFU/mL×10⁶(81.266 7+3.341 8), CFU/mL(90.900 0+2.040 1)×10⁶,CFU/mL(50.866 7+2.208 6)×10⁶, CFU/mL(77.666 7+2.880 7)×10⁶ CFU/mL, colony counting of blank group was(92.033 3+3.890 6)×10⁶ CFU/mL, respectively. Compared with the blank group, 1/2MIC Coptis decoction, 1/4MIC Coptis decoction,1/8MIC Coptis decoction,1/2MIC Oxacillin sodium,1/4MIC Oxacillin sodium all had differences in statistical significantce(P=0.001), but 1/16MIC Coptis decoction had no statistically significant(P=0.173). In the BF model after 7 days, colony counting bacteria, crystal violet and silver staining showed similar results. Further studies showed that under the effect of 1/16MIC Coptis decoction, the minimum inhibitory concentration of oxacillin sodium was 0.25 μg/mL. Conclusion Coptis decoction could inhibit the formation of Staphylococcus aureus biofilm, and oxacillin sodium combined with Coptidis decoction was effective to improve the curative effect of oxacillin sodium, providing new treatment for peritoneal dialysis related peritonitis that is caused by Staphylococcus aureus improving the efficacy of antibiotics, and providing help for the clinical on reducing the generation of antibiotic resistant bacteria.

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