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

doi:10.3969/j.issn.1000-8535.2018.01.004

广州医药 ›› 2018, Vol. 49 ›› Issue (1): 13-17.DOI: 10.3969/j.issn.1000-8535.2018.01.004

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

李奕铮, 潘玲, 廖蕴华

广西医科大学第一附属医院肾内科(南宁 530021)

收稿日期:2017-10-01 出版日期:2018-01-20 发布日期:2021-12-01
通讯作者: 廖蕴华,E-mail:yunhualiao1989@sina.com
基金资助:
广西中医药管理局中医药自筹课题(GZZC15-52);国家自然科学基金(81360111)

Study on the effect of Coptis chinensis decoction on the biofilm of Staphylococcus aureus and its effects on oxacillin sodium

LI Yizheng, PAN Ling, LIAO Yunhua

Department of Nephrology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021,China

Received:2017-10-01 Online:2018-01-20 Published:2021-12-01

摘要/Abstract

摘要： 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。

关键词: 黄连水煎剂, 金黄色葡萄球菌, 生物膜, 苯唑西林钠

Abstract: 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.

Key words: Coptis decoction, Staphylococcus aureus, Biofilm, Oxacillin sodium

李奕铮, 潘玲, 廖蕴华. 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[J]. 广州医药, 2018, 49(1): 13-17.

LI Yizheng, PAN Ling, LIAO Yunhua. Study on the effect of Coptis chinensis decoction on the biofilm of Staphylococcus aureus and its effects on oxacillin sodium[J]. Guangzhou Medical Journal, 2018, 49(1): 13-17.

参考文献

[1] 王悦. 腹膜透析相关并发症分析[D]. 广州:南方医科大学,2016.
[2] 林爱武,钱家麒. 腹膜透析管出口感染的菌种和预后分析[J]. 中华肾脏病杂志,2003,19(2):11-13.
[3] 杜仲业,陈一强,孔晋亮,等. 黄芩苷对金黄色葡萄球菌生物膜抑制作用的体外研究[J]. 中华医院感染学杂志,2012(8):1541-1543.
[4] NADELL C D, DRESCHER K, FOSTER K R.Spatial structure, cooperation and competition in biofilms[J]. Nat Rev Microbiol, 2016,14(9):589-600.
[5] FLEMMING H C, WINGENDER J, SZEWZYK U. et. al. Biofilms: an emergent form of bacterial life[J]. Nat Rev Microbiol,2016,14(9):563-575.
[6] 胡晓丰,史云,戚丽华,等. 金黄色葡萄球菌生物膜形成机制研究进展[J]. 生物技术通讯,2014,25(5):714-718.
[7] 施燕春,项新. 腹膜透析患者相关炎性因子的研究进展[J]. 医学综述,2016,22(12):2310-2313.
[8] Tăranu T, FLOREA L, Păduraru D, et al. Morphological changes of the peritoneal membrane in patients with long-term dialysis[J]. Rom J Morphol Embryol, 2014,55(3): 927-932.
[9] 周巍,王葳,韩国锋,等. 腹膜透析患者的转归及其危险因素分析[J]. 中国血液净化,2012,11(11):600-605.
[10] HALL C W, MAH T F. Molecular mechanisms of biofilm-based antibiotic resistance and tolerance in pathogenic bacteria[J]. FEMS Microbiol Rev, 2017,41(3):276-301.
[11] BUOMMINO E, SCOGNAMIGLIO M, DONNARUMMA G, et al. Recent advances in natural product-based anti-biofilm approaches to control infections[J]. Mini Rev Med Chem, 2014,14(14):1169-1182.
[12] FERREIRA M,RZHEPISHEVSKA O,GRENHO L, et al. Levofloxacin-loaded bone cement delivery system: highly effective against intracellular bacteria and Staphylococcus aureus biofilms[J]. Int J Pharm, 2017,532(1):241-248.
[13] YAN J, NADELL C D, BASSLER B L. Environmental fluctuation governs selection for plasticity in biofilm production[J]. ISME J, 2017,11(7):1569-1577.
[14] MARTIN M, Dragoš A, Hölscher T, et al. De novo evolved interference competition promotes the spread of biofilm defectors[J]. Nat Commun,2017(8):15127.
[15] LIU J, MARTINEZCORRAL R, PRINDLE A,et al. Coupling between distant biofilms and emergence of nutrient time-sharing[J]. Science,356(6338):638-642.
[16] DALCIN A J F, SANTOS C G, Gündel S S, et al. Anti biofilm effect of dihydromyricetin-loaded nanocapsules on urinary cathete.r infected by Pseudomonas aeruginosa[J]. Colloids Surf B Biointerfaces, 2017(156):282-291.
[17] 彭齐,谢丽玲,谢俊. 中药抑菌机制的研究方法[J]. 生物技术进展,2015,5(1):66-69.
[18] 胡淑平,王怡,于平野,等. 黄连解毒汤主要成分的体外抗炎作用研究[J]. 中国现代应用药学,2014,31(10):1171-1174.
[19] 肖丽英,黄焯坡. 23种中草药对耐药金葡菌的敏感性探讨[J]. 时珍国医国药,2001,12(10):878-879.

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

Study on the effect of Coptis chinensis decoction on the biofilm of Staphylococcus aureus and its effects on oxacillin sodium

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 4

编辑推荐

Metrics

本文评价

[1]	梁慧玲, 韩超, 郑琳颖, 张秋玲, 刘道利, 周伟青. 翘芩清肺剂中药免煎颗粒剂与饮片剂的抑菌作用比较[J]. 广州医药, 2021, 52(3): 112-116.
[2]	林颜玉, 陈文伟, 王柏旺, 卢玉平. 中山市部分地区金黄色葡萄球菌感染的临床分布及耐药基因研究[J]. 广州医药, 2016, 47(2): 7-9.
[3]	梁志科, 赵子文, 刘朝晖, 李裕军, 钟维农. 磷酸二酯酶4抑制剂对人巨噬细胞噬菌能力的影响[J]. 广州医药, 2015, 46(5): 5-8.
[4]	李敏豪, 董超, 王成兴, 董婉华, 冯素娥, 刘凤鸣. 广州地区儿童社区获得性耐甲氧西林金黄色葡萄球菌分子特征研究[J]. 广州医药, 2015, 46(3): 25-27.