CN117398430A - 鸡矢藤和含鸡矢藤提取物的组合物及其应用 - Google Patents
鸡矢藤和含鸡矢藤提取物的组合物及其应用 Download PDFInfo
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- CN117398430A CN117398430A CN202310988072.0A CN202310988072A CN117398430A CN 117398430 A CN117398430 A CN 117398430A CN 202310988072 A CN202310988072 A CN 202310988072A CN 117398430 A CN117398430 A CN 117398430A
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- fevervine
- mrsa
- staphylococcus aureus
- gentamicin
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Abstract
本发明涉及鸡矢藤和含鸡矢藤提取物的抗菌组合物及其应用。本发明提供了包括抗生素和鸡矢藤提取物的用于预防和/或治疗黄色葡萄球菌感染的组合物在制备预防和治疗金黄色葡萄球菌感染的制剂中的应用。本发明通过大量研究,意外发现鸡矢藤针对临床多重耐药MRSA菌株具备显著抗菌活性,其能通过抑制MRSA对数生长期的增殖而发挥抗MRSA作用;鸡矢藤提取物的抑菌机理可能是通过阻碍菌体某些蛋白的合成,从而起到抑制细菌生长的作用。并且,试验结果表明,鸡矢藤提取物与庆大霉素联合用药时还可产生协同作用。本发明将鸡矢藤提取物作为庆大霉素增效剂甚至于在制备抗菌组合物中,具备广阔的临床应用及发展前景。
Description
技术领域
本发明涉及生物医药领域,具体涉及鸡矢藤和含鸡矢藤提取物的组合物及其应用。
背景技术
耐药菌感染已成为医疗机构面临的一个严峻挑战,严重威胁人类健康。其中耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)是临床上常见的耐药菌,随着抗生素的滥用,细菌耐药率呈现逐年上升趋势,MRSA在临床的检出率越来越高,且呈多重耐药趋势,全国细菌耐药监测网及CHINET的监测资料显示多重耐药菌MRSA的感染率在50%以上,且ICU、烧伤病房中,感染有加重的趋势。由于MRSA具有多重耐药性,临床中面临治疗难度高、死亡率高等困难,因此成为世界范围内耐药性检测的重点对象。目前,临床上可用于治疗MRSA的药物非常有限,一线药物主要有万古霉素、替考拉宁和去甲万古霉素等抗生素,而这些药物价格昂贵,且具有耳毒性和肾毒性大等缺点,临床用药时需谨慎选择。因此,开发更多经济有效的抗MRSA药物是目前需要迫切解决的问题。而随着新抗生素的研发也陷入瓶颈,因此拓宽抗菌药物的来源渠道是一条重要举措。中药具有多功能、多靶点作用、不易产生耐药性、不良反应低以及与抗生素联用可逆转细菌耐药性等优点,且上千年的实践证明中医药在治疗难治性感染疾病上具有良好的疗效。近年来,已发现多种中药及相关活性成分具有显著抗耐药菌作用,从中药中开发抗菌药物将形成一个新的研究领域。
对于天然药物体外、体内和体内外联合抑菌活性的试验方法而言,不同类别天然药物的适用方法各异,因此,需要采用适当的方法,筛选出适合的针对特定菌群或耐药菌的中药或天然药物,从而针对细菌或耐药菌起到抑菌或杀菌作用。
鸡矢藤性平,味甘、微苦,具有祛风利湿、止痛解毒、活血消肿及镇痛和抗炎作用,临床上多用于多种疼痛和胃肠道疾病的治疗,除此之外还可用于带状疱疹、结核、痈疮肿毒和蛇虫咬伤等疾病的治疗,具有很高的药用价值。随着临床的发展和研究,对其更多的作用也在开发中,以期望获得更多对人类有益的用途。
发明内容
基于此,本发明的目的在于提供鸡矢藤或鸡矢藤提取物在医学中的新应用,所述鸡矢藤或鸡矢藤提取物单独或者与抗生素联合使用,能够用于预防和治疗金黄色葡萄球菌感染的,特别是能够对耐药金黄色葡萄球菌起到抑菌或杀菌作用。
本发明的第一个方面,是提供了鸡矢藤或鸡矢藤提取物联合抗生素在制备预防和治疗金黄色葡萄球菌感染的制剂中的应用,所述抗生素选自硫酸多粘菌素B、左氧氟沙星、庆大霉素、万古霉素、氨苄青霉素中的至少一种。
本发明中,所述鸡矢藤提取物可以为本领域技术人员常规所知的任一种提取方法获得的相应提取物。
在其中一些实施例中,所述鸡矢藤提取物为鸡矢藤的水提物或者醇提物。
在其中一些实施例中,所述鸡矢藤提取物为鸡矢藤通过乙醇联合超声方式的提取物。
本发明的第二个方面,是提供了一种预防和/或治疗黄色葡萄球菌感染的组合物,其活性成分包括抗生素和鸡矢藤提取物,所述抗生素和所述鸡矢藤提取物分别成为独立的给药单元,或所述抗生素和所述鸡矢藤提取物共同形成组合的给药单元。
在其中一些实施例中,所述抗生素包括但不限于硫酸多粘菌素B、左氧氟沙星、硫酸庆大霉素、万古霉素、氨苄青霉素;优选地,所述抗生素为庆大霉素。
在其中一些实施例中,所述鸡矢藤提取物采用醇提联合超声提取的方式制备而成。
在其中一些实施例中,所述鸡矢藤提取物包括:取干燥鸡矢藤全草,经粉碎后,加入8-12倍量的50%-70%的乙醇水溶液,40℃-50℃超声提取1-2小时,期间搅拌多次;过滤后得到的药渣再加8-12倍量的50%-70%的乙醇水溶液加热1-2小时,合并滤液,浓缩后即得。
本发明的第三个方面,是提供了上述任一种组合物在制备用于抗金黄色葡萄球菌制剂中的应用。
在其中一些实施例中,所述制剂为抗生素与鸡矢藤提取物共同形成组合的给药单元,且还可包含药学上可接受的载体、溶剂、稀释剂、赋形剂或其他介质。
在其中一些实施例中,所述金黄色葡萄球菌为耐药菌株,进一步地,所述耐药菌株为多重耐药菌株;更优选地,所述多重耐药菌株为MRSA。
本发明在进行中药抗病原微生物研究时,意外地发现中药鸡矢藤提取物对30株MRSA临床多重耐药株均具有不同程度(MIC为62.5-250mg/mL,MBC为250-500mg/mL)的抗菌效果,结合动态抑菌试验和SDS-PAGE测定可溶性总蛋白变化等试验表明,不同浓度(1/4×MIC-2×MIC)鸡矢藤提取物后MRSA菌体生长曲线均发生明显变化,其能通过抑制MRSA对数生长期的增殖而发挥抗MRSA作用。对MRSA抑制作用的强弱与鸡矢藤提取物浓度呈正相关,结果表明鸡矢藤提取物的抑菌机理可能阻碍菌体某些蛋白的合成,从而起到抑制细菌生长的作用。
同时,值得关注的是,鸡矢藤提取物与庆大霉素联合用药可协同抗MRSA,显著降低庆大霉素的MIC,增加MRSA对庆大霉素的敏感性,鸡矢藤提取物协同庆大霉素抑制MRSA,能够有效减少庆大霉素的用量,降低其毒副反应。虽其具体协同作用机制亟待进一步研究,但迄今为止,鸡矢藤提取物提高氨基糖苷类抗生素对金葡菌的抗菌作用均未见报道。充分表明鸡矢藤提取物作为庆大霉素增效剂甚至于在制备抗菌组合物中,具备广阔的临床应用及发展前景。
附图说明
图1为实施例1中药敏试验部分结构代表图,其中A-D分别为耐甲氧西林金黄色葡萄球菌M64上各药敏纸片检测结果;E-H分别为耐甲氧西林金黄色葡萄球菌M66上各药敏纸片检测结果。
图2为实施例1中97株MRSA临床分离株的多重耐药率统计图。
图3为实施例2中鸡矢藤提取物采用牛津杯法进行抑菌试验的结果图,其中A-H分别为鸡矢藤对ATCC25923、ATCC43300、M29、M30、M37、M39、M43、M46的牛津杯抑菌结果;其中编号1、4、7、10、13、16、19、22为鸡矢藤提取液原液;2、5、8、11、14、17、20、23为浓度1/2的鸡矢藤提取液;3、6、9、12、15、18、21、24为浓度1/4的鸡矢藤提取液;C为阳性对照:盐酸左氧氟沙星氯化钠注射液(海力健,批号80120051202);NC为阴性对照:无菌水。
图4为实施例3中采用微量稀释法测定鸡矢藤提取物对MRSA的MIC结果。
图5为实施例4中鸡矢藤提取物抗MRSA的动态抑菌曲线,其中试验菌株为MRSA临床株M3。MIC:鸡矢藤提取物62.5mg/mL。
图6为实施例5中鸡矢藤提取物对MRSA总蛋白含量变化的检测结果图,其中M为蛋白Marker。
图7为实施例6中鸡矢藤提取物与庆大霉素针对ATCC43300采用棋盘法检测抗菌效果图。
具体实施方式
下列实施例中未注明具体条件的实验方法,通常按照常规条件,例如Sambrook等人,分子克隆:实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件,或按照制造厂商所建议的条件。实施例中所用到的各种常用化学试剂,均为市售产品。
除非另有定义,本发明所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不用于限制本发明。本发明所使用的术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。
在整个说明书和权利要求书中,以下术语具有与本文明确相关的含义,除非上下文另有明确规定。在本发明中使用的短语“在一个实施方案中”不一定指代相同的实施方案,尽管其可能是。此外,在本发明中使用的短语“在另一实施方案中”不一定指代不同的实施方案,尽管其可能是。因此,如下所述,可以容易地组合本发明的各个实施方案,而不脱离本发明的范围或精神。
为了便于理解本发明,下面将对本发明进行更全面的描述。本发明可以以许多不同的形式来实现,并不限于本文所描述的实施例。相反地,提供这些实施例的目的是使对本发明公开内容的理解更加透彻全面。
以下结合具体实施例对本发明作进一步详细的说明。
实施例1临床菌株耐药谱的测定
试验菌株为97株耐甲氧西林金黄色葡萄球菌临床分离株(MRSA)由中山大学附属第三医院岭南医院馈赠,由该医院2020年1月-12月临床分离所得,来源包括患者痰液、分泌物、脓液、血液、尿液和脑脊液等,经梅里埃公司VITEK 2Compact鉴定。质控菌株为金黄色葡萄球菌标准菌株ATCC25923购于环凯生物。
药敏试验严格参照《全国临床检验操作规程》(第3版)进行。药敏试验使用纸片琼脂扩散法(即K-B法)检测MRSA对不同抗菌药物的敏感性。将稀释后的菌液用无菌棉签蘸取均匀涂布于M-H琼脂平板上,待干后,分别贴上药敏纸片(均购自Oxoid公司,分别为P(青霉素)、E(红霉素)、TE(四环素)、VA(万古霉素)、DA(克林霉素)、KZ(头孢唑啉)、SXT(复方新诺明)、TGC(替加环素)、CIP(环丙沙星)、LZD(利奈唑胺)、CN(庆大霉素)、FOX(头孢西丁)、OX(苯唑西林)),正面放置30min后,倒置于35℃培养箱培养18h,测定抑菌环大小,根据CLSI2013年M100-S23的耐药性标准,判断其对药物的耐药性。
药敏试验结果统计表及部分结果图分别如下比表1和图1所示,其中上述97株临床MRSA对青霉素(94.85%)和红霉素(90.72%)的耐药率均高于90.00%,对苯唑西林(77.32%)、头孢西丁(68.04%)、克林霉素(65.98%)、环丙沙星(55.67%)、四环素(51.55%)和头孢唑林(50.52%)的耐药率也较高,均超过50%,对复方磺胺甲噁唑、利奈唑烷、万古霉素几乎不耐药(RR≤5.00%)。
表1.K-B法测定临床分离MRSA对常见抗生素的耐药率
进一步地,根据多重耐药指数MARI=a/bc(a为细菌的总抗菌药物耐药值,b为抗菌药物种类数目,c代表细菌菌株数;0<MARI<1,MARI值越大说明多重耐药水平越高)计算公式,应用SPSS软件进行统计分析,使用GraphPad Prism作图,临床菌株耐药率统计图如图2所示,97株MRSA的临床分离株中,78株表现出多重耐药菌特征(显示出对三类及以上抗生素显著耐药性),占80.41%,多重耐药指数MARI为0.5069。
实施例2牛津杯法检测鸡矢藤提取物的抑菌作用
1.材料
鸡矢藤提取物的制备:精密称取100g鸡矢藤碾碎物,加入10倍量的60%乙醇,采用45℃超声提取90min,期间搅拌多次;过滤后弃去滤渣,滤液经旋转蒸发仪浓缩后加水定容至100ml;用0.22μm微孔滤膜过滤除菌后作为药物原液(1g生药量/mL),分装使用。
阳性对照药物:盐酸左氧氟沙星氯化钠注射液(海力健,批号80120051202)、硫酸多粘菌素B(Aladdin,批号J2126657)、左氧氟沙星(Aladdin,批号J2122423)、硫酸庆大霉素(Aladdin,批号H2106059)、万古霉素(Aladdin,批号H2125431)、氨苄青霉素(Aladdin,批号E2011092)。
阳性对照药物的预处理方法:取适量多粘菌素B采用生理盐水稀释混匀至终浓度为10mg/mL的工作液;取适量的左氧氟沙星和氨苄青霉素,分别采用生理盐水稀释混匀至终浓度为2mg/mL的工作液;取适量庆大霉素采用生理盐水稀释混匀至终浓度为1mg/mL的工作液;取适量的万古霉素采用生理盐水稀释混匀至终浓度为0.2mg/mL的工作液。
试验菌株:56株随机从上述实施例1中78株表现出多重耐药菌的MRSA临床分离株(MRSA)。
质控菌株:金黄色葡萄球菌标准菌株ATCC25923和MRSA标准菌株ATCC43300,均购于环凯生物。
2.方法
分别将金黄色葡萄球菌标准菌株ATCC25923、MRSA标准菌株ATCC43300以及56株MRSA临床菌株接种于MH肉汤培养基中,于恒温空气浴震荡培养箱中以37℃、200r/min振荡培养12h。将上述菌液在MH琼脂培养基上划线培养,挑单克隆菌落于MH肉汤培养基中,以37℃、200r/min振荡培养,完成菌种的活化。活化后的菌液以8000r/min离心5min,弃掉上清,再用灭菌的MH肉汤培养基稀释菌体沉淀至1×106CFU/mL,作为实验菌液。
先将已灭菌的MH琼脂培养基倒入培养皿,待其凝固。此外,待MH琼脂培养基冷却至45℃左右混入相应实验菌液5μl,混合均匀后准确分取混菌培养基5.0mL完全覆盖在已凝固的培养基上并保证其表面光滑,等待凝固。取已高压灭菌的牛津杯置于培养基表面,在杯中加入不同浓度的鸡矢藤提取物,以灭菌双蒸水作阴性对照,左氧氟沙星工作液为阳性对照,将平板置于恒温培养箱中37℃下培养过夜,将平板取出后。采用“十字交叉法”测定抑菌圈直径,拍照后记录相关结果,以抑菌环直径>7mm判断为有抑菌作用。检测结果分别如下表2、图3所示。
表2鸡矢藤提取物的MRSA临床株抑菌效果统计
分别对金黄色葡萄球菌ATCC25923、MRSA标准菌株ATCC43300和56株MRSA临床株进行鸡矢藤提取物的抑菌效果评价,牛津杯抑菌试验结果显示,鸡矢藤提取物对ATCC25923、ATCC43300及其中30株MRSA临床株均具有不同程度的抑菌活性(抑菌环直径菌>7mm),对MRSA临床株抑菌率占53.57%(30/56),见图3及表2。由图3可见,鸡矢藤提取物浓度为500mg/ml和250mg/ml时均能表现出显著的抗菌活性,且MRSA对不同浓度的鸡矢藤提取物的敏感性呈一定的浓度依赖关系。综合结果表明,鸡矢藤提取物对MRSA具有一定的抑菌作用。
实施例3鸡矢藤提取物对MRSA的MIC和MBC测定
采用倍比稀释法将鸡矢藤提取物以及5种抗生素(多粘菌素B、左氧氟沙星、庆大霉素、氨苄青霉素、万古霉素)进行稀释,从1:2倍稀释至1:512倍。倍比稀释后对应的鸡矢藤提取物浓度分别为500、250、125、62.5、31.25、15.625、7.813和3.906mg/mL(1/2-1/256)。分别在96孔板中先后加入100μL不同浓度的稀释药液以及抗生素,然后加入1×106CFU/mL MRSA菌悬液100μL,同时做空白对照,于37℃恒温培养18h,观察受试菌的生长情况(肉眼观察浑浊度),于各孔中加入20μL 0.2%的TTC,避光,培养4h,实验重复3次,取平均值,以不加药物组为空白对照。没有红色还原物甲臢产生的最低药物浓度为最低抑菌浓度(minimuminhibitory concentration,MIC)。测出MIC后,再依次从无细菌生长的各孔中吸取100μL培养物分别接种于营养琼脂平板,置于恒温培养箱37℃下培养18~24h,平板上生长的菌落数<5个对应的药液浓度为最低杀菌浓度(MBC)。
表3鸡矢藤提取物抗MRSA的MIC和MBC测定结果
注:“+”培养基浑浊;“-”培养基澄清
微量稀释法结果显示,鸡矢藤提取物对金黄色葡萄球菌ATCC25923和MRSA标准菌ATCC43300的MIC均为125mg/mL,MBC均为500mg/mL,对MRSA临床分离株的MIC为62.5-250mg/mL,MBC为250-500mg/mL(如图4,表3所示)。
实施例4鸡矢藤提取物对MRSA生长曲线的影响
将MRSA活化后用生理盐水配制成0.5个麦氏浊度备用。实验分为阳性对照组(仅加菌液不加药液)、鸡矢藤提取物(含1/4×MIC、1/2×MIC、1×MIC及2×MIC)组。以不加菌液仅加相同浓度药液的为阴性对照组,用于消减药物颜色干扰。在含MH液体培养液中分别加入1%菌液量和对应药液,37℃、180r/min振荡培养24h,每2小时无菌取样1次,用酶标仪测定600nm波长的吸光度(A),其中实验组OD600为实验处理组与同浓度同时段阴性组的吸光度之差,取3次重复实验的平均值。以t为横坐标(X),A600为纵坐标(Y),绘制MRSA生长曲线,分析不同浓度鸡矢藤提取物对MRSA生长的影响。应用SPSS软件进行统计分析,使用GraphPadPrism作图。
鸡矢藤提取物对MRSA生长抑制曲线如图5所示。与对照组相比,经鸡矢藤提取物处理的1/4×MIC、1/2×MIC、1×MIC和2×MIC组均出现了不同程度的抑制生长趋势,细菌溶液的光密度显著降低,且鸡矢藤提取物浓度越高对细菌生长的影响越大。从生长曲线的变化来看,鸡矢藤提取物缩短MRSA的对数生长期,延长生长停滞期,导致细菌数量明显减少。结果表明,鸡矢藤提取物对MRSA的正常生长和繁殖具有抑制作用,且抑制作用的强弱与鸡矢藤提取物浓度呈正相关。
实施例5鸡矢藤提取物对MRSA总蛋白含量变化的影响
制样:向30mL MH培养基中加入1%(v/v)培养至对数期的MRSA。在进行增菌培养后,加入不同浓度的鸡屎藤提取物(1/16×MIC~2×MIC),以不加鸡屎藤提取物而加相同体积的无菌水作为空白组。37℃下培养4h后,低温离心收集100mg菌体,悬于100μL无菌水中,并加入25μL 4×SDS-PAGE Loading Buffer。将样品沸水浴8min,8500rpm离心5min,取上清液进行SDS-PAGE电泳。
蛋白质具有维持集体新陈代谢及各类物质在体内的输送等功能,是生命活动的主要承担者。如果药物使菌体蛋白质特定的空间结构遭到破坏,其生物功能将丧失,药物将起到抑菌作用。由图6可知,与对照组相比,鸡矢藤提取物作用后,MRSA可溶性蛋白含量明显降低,但是蛋白种类未发生明显改变,且随着鸡矢藤提取物的浓度增加,对应实验组的MRSA可溶性蛋白条带变浅,表明蛋白含量降低,即对MRSA抑制作用的强弱与鸡矢藤提取物浓度呈正相关,结果表明鸡矢藤提取物的抑菌机理可能阻碍菌体某些蛋白的合成,从而起到抑制细菌生长的作用。
实施例6鸡矢藤提取物与抗生素联合用药的作用效果
根据鸡矢藤提取物和4种常用抗生素的MIC结果(参考实施例2检测方法,检测结果如表5所示),采用倍比稀释法分别稀释使其终浓度分别1/32MIC~4MIC。采用微量棋盘法稀释法测定鸡矢藤提取物分别与抗生素(多粘菌素B、左氧氟沙星、庆大霉素、氨苄青霉素和万古霉素)联用的MIC,分析其FIC。其中,FIC指数=联合用药时甲药MIC/单独应用甲药时MIC+联合应用乙药时MIC/单独应用乙药时MIC。判断标准:FIC指数≤0.5为协同作用,0.5<FIC指数≤1为相加作用,1<FIC指数≤2为无关作用,FIC指数>2为拮抗作用。以鸡矢藤提取物与庆大霉素联合抑制M3的试验为例:
将鸡矢藤提取物和庆大霉素的工作液稀释成4倍MIC(即鸡矢藤1/4;庆大霉素1/8)。96孔板每排从左向右编号1-12,从上向下编号A-H。
1)在9A-9G加入鸡矢藤,在12A-12H加入抗生素各100μL。
2)在2A-8G加入50μL培养基。
3)将第9列的鸡矢藤吸50μL到第8列,反复吹打3次,再吸50μL到第7列,依次类推到第2列,最后50μL弃掉。
4)将第12列的抗生素吸50μL到G行,使之满100μL,再吸培养基50μL在第10列,将抗生素倍比稀释后吸取50μL到F行,使之满100μL,依次类推至A行。
5)在10A加入200μL鸡矢藤,在11A加入200μL抗生素,10B和11B以下加入100μL培养基,将10A和11A依次倍比稀释。
6)加入110μL的M3菌悬液,放入37℃培养箱中过夜培养16-20小时,取出后观察细菌生长情况并记录结果,并于各孔中加入20μL 0.2%的TTC,避光,培养4h,实验重复3次,取平均值,以不加药物组为空白对照。
表4.棋盘法联合药敏示意表
表5.抗生素和鸡矢藤提取物对试验菌株的MIC
表6.鸡矢藤提取物与抗生素联用对试验菌株的FIC
表7.鸡矢藤联合抗生素前后对试验菌株的MIC变化
通过棋盘法测定鸡矢藤提取物与不同抗生素之间的联合用药抑菌效果,结果如表6、7所示,其中,鸡矢藤提取物可显著提高庆大霉素(FIC=0.375)对ATCC43300的抑菌作用,联合使用呈现出协同效果(如图7所示,96孔板中的D5孔澄清,且经添加TTC培养后不变红,表示该孔未长菌,即鸡矢藤提取物在1/8MIC浓度时,可将庆大霉素的MIC从1/16降低至1/4×1/16=1/64)。
此外,一定程度上提高左氧氟沙星(FIC=0.625)、氨苄青霉素(FIC=0.531)和万古霉素(FIC=0.531)对ATCC43300的抑菌作用,联用呈相加作用,与多粘菌素B联用对ATCC43300呈无关作用。另外,鸡矢藤提取物联用多粘菌素B(FIC=0.625)、氨苄青霉素(FIC=1.0)、左氧氟沙星(FIC=0.531)和庆大霉素(FIC=0.75)对MRSA临床菌株M3呈相加作用,而与万古霉素联用对MRSA临床菌株呈无关作用,且联合左氧氟沙星(FIC=1.0)、庆大霉素(FIC=0.516)和万古霉素(FIC=0.625)对金黄色葡萄球菌ATCC25923均有相加的抑菌作用。
本实验结果显示,鸡矢藤提取物与庆大霉素联合用药可协同抗MRSA标准菌株ATCC43300,显著降低庆大霉素的MIC,增加ATCC43300对庆大霉素的敏感性,还能一定程度提高对金黄色葡萄球菌ATCC25923和MRSA临床株的抑菌作用,联用呈现出相加作用,但具体的协同、相加作用机制需进一步研究。庆大霉素是氨基糖苷类抗生素,可通过与细菌核糖体30S亚基结合,阻断细菌蛋白质合成从而发挥抗菌作用,具有较强的耳、肾毒性。鸡矢藤提取物协同庆大霉素抑制ATCC43300,能够有效减少庆大霉素的用量,降低其毒副反应。且迄今为止,鸡矢藤提取物提高氨基糖苷类抗生素对金葡菌的抗菌作用均未见报道。因此,鸡矢藤提取物作为庆大霉素增效剂具有潜在的开发价值。
此外,鸡矢藤提取物与多粘菌素B、氨苄青霉素、左氧氟沙星和万古霉素联合用药后对金黄色葡萄球菌ATCC25923和MRSA的作用效果均没有十分显著的提高,其中联合多粘菌素B、氨苄青霉素、左氧氟沙星对MRSA临床菌株,联合氨苄青霉素、左氧氟沙星和万古霉素对MRSA标准株ATCC43300以及联合左氧氟沙星和万古霉素对金黄色葡萄球ATCC25923均表现为相加作用,显示出一定的抑菌效果以及联合用药有效性,且结果中联合用药均无出现拮抗作用,说明鸡矢藤提取物与以上抗生素配伍使用可行。虽然目前研究所用药物为鸡矢藤的粗提物,具体的有效成分及其抑菌机制有待进一步深入研究。以上结果均提示着鸡矢藤提取物的天然抗菌成分可能为解决抗生素耐药菌急剧增多的问题开辟新的领域。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (10)
1.鸡矢藤或鸡矢藤提取物联合抗生素在制备预防和治疗金黄色葡萄球菌感染的制剂中的应用,所述抗生素选自硫酸多粘菌素B、左氧氟沙星、庆大霉素、万古霉素、氨苄青霉素中的至少一种。
2.根据权利要求1所述的应用,其特征在于,所述鸡矢藤提取物为鸡矢藤的水提物或者醇提物。
3.根据权利要求1或2所述的应用,其特征在于,所述金黄色葡萄球菌为耐甲氧西林金黄色葡萄球菌。
4.一种预防和/或治疗金黄色葡萄球菌感染的组合物,其特征在于,所述组合物的活性成分包括抗生素和鸡矢藤提取物,所述抗生素和所述鸡矢藤提取物分别成为独立的给药单元,或所述抗生素和所述鸡矢藤提取物共同形成组合的给药单元。
5.根据权利要求3所述的组合物,其特征在于,所述抗生素选自硫酸多粘菌素B、左氧氟沙星、庆大霉素、万古霉素、氨苄青霉素中的至少一种。
6.根据权利要求5所述的组合物,其特征在于,所述抗生素为庆大霉素。
7.根据权利要求4-6任一项所述的组合物,其特征在于,所述鸡矢藤提取物为鸡矢藤的水提物或者醇提物。
8.根据权利要求7所述的组合物,其特征在于,所述鸡矢藤提取物为醇提物,其制备方法包括:取鸡矢藤全草,经粉碎后,加入8-12倍量的50%-70%的乙醇水溶液,40℃-50℃超声提取1-2小时,期间搅拌多次;过滤得滤液。
9.权利要求4-8任一项的组合物在制备预防和治疗金黄色葡萄球菌感染的制剂中的应用。
10.根据权利要求9所述的应用,其特征在于,所述金黄色葡萄球菌为耐药菌株;进一步地,所述金黄色葡萄球菌为耐甲氧西林金黄色葡萄球菌。
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