CN110946863A - 白屈菜红碱在抑制路邓葡萄球菌生长中的应用 - Google Patents
白屈菜红碱在抑制路邓葡萄球菌生长中的应用 Download PDFInfo
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- staphylococcus lugdunensis
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- 241001134656 Staphylococcus lugdunensis Species 0.000 title claims abstract description 60
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- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract
本发明公开了白屈菜红碱在抑制路邓葡萄球菌生长中的应用,根据白屈菜红碱对耐药路邓葡萄球菌具有较好的体外杀灭作用,能够抑制耐药路邓葡萄球菌的生长,且最低杀菌浓度为16μg/mL,最小抑菌浓度为7.8μg/mL,本发明提出了白屈菜红碱对路邓葡萄球菌的抑制作用,在医药等领域具有广泛的应用价值。
Description
技术领域
本发明涉及中药有效成分白屈菜红碱作为路邓葡萄球菌的抑制剂的研究开发和应用,具体涉及白屈菜红碱在抑制路邓葡萄球菌生长中的应用。
背景技术
路邓葡萄球菌(Staphylococcus lugdunensis)是凝固酶阴性的葡萄球菌之一,喜寄殖于会阴部皮肤。路邓葡萄球菌也可引起医疗操作相关性感染,其中最常见的为感染性心内膜炎(IE),包括原发性IE、人工瓣膜置换术后IE及起搏器相关性IE。另外,路邓葡萄球菌还可引起腹膜炎、泌尿道感染、中枢神经系统感染、骨、关节感染(脓毒性关节炎、骨髓炎)、眼部感染(化脓性角膜炎、术后眼内炎)、口腔感染等。随着抗生素在临床上的广泛使用,细菌的耐药性也越来越强。虽然目前路邓葡萄球菌的耐药现状良好,但却出现了对糖肽类抗菌药物耐药的个别报道,也有报道指出路邓葡萄球菌对青霉素已经产生了耐药性,对阿奇霉素、氯霉素和庆大霉素的敏感性也逐渐降低。Bourgeois等对90株凝固酶阴性葡萄球菌耐药情况的研究发现,路邓葡萄球菌对万古霉素及替考拉宁的敏感性降低。
中药资源无论在感染控制还是在现代医疗方面都凸显出独特的优势,中药取药天然,起效方式全面,近年来,中草药抑菌作用的药理学研究及中药成方的临床应用也越来越多地受到学者们的关注。白屈菜红碱是从拟花椒、马钱子等罂粟科植物根部中分离得到的生物碱,具有多种药理学作用,如抗肿瘤、抑菌、抗炎作用等。
文献(范海延,薛广厚,刘晶瑜,et al.白屈菜红碱的分离纯化及其抑菌活性[J].湖北农业科学,2010,49(3):679-682.)采用大孔吸附树脂分离纯化白屈菜中的白屈菜红碱,并对白屈菜红碱的抑菌效果进行了研究。结果表明,纯化后的白屈菜红碱对甜瓜枯萎病病菌、西瓜枯萎病病菌、茄子黄萎病病菌、黄瓜枯萎病病菌、辣椒炭疽病病菌的菌丝生长均具一定抑制作用。中国专利报道了“白屈菜红碱在治疗禽耐药性大肠杆菌病上的应用”(CN106138054A),其中,大肠杆菌为肠杆菌科埃希氏菌属。但是未见关于白屈菜红碱抑制路邓葡萄球菌生长方面的报道。
发明内容
本发明针对目前临床医学面临的耐药难题,目的在于提供白屈菜红碱在抑制路邓葡萄球菌生长中的应用。
为了实现上述目的,本发明采用的技术方案为:
利用纸片扩散法确定路邓葡萄球菌对抗生素的耐药性,通过微量二倍稀释法确定白屈菜红碱对路邓葡萄球菌的最小抑菌浓度(MIC)和最低杀菌浓度(MBC)。结果表明:白屈菜红碱对耐药路邓葡萄球菌具有较好的抑菌作用,可以用于抑制耐药路邓葡萄球菌的生长。
优选的,所述路邓葡萄球菌为耐青霉素、克林霉素、四环素中一种以上抗生素的人源路邓葡萄球菌。
优选的,所述路邓葡萄球菌选自耐青霉素、克林霉素和四环素的人源路邓葡萄球菌。
优选的,所述白屈菜红碱的最低杀菌浓度为16μg/mL,最小抑菌浓度为7.8μg/mL。
本发明的有益效果体现在:
本发明从现存的药用植物资源库入手,挖掘潜在的耐药菌抑制剂,基于白屈菜红碱对耐药路邓葡萄球菌的作用的研究,发现非抗生素类化合物白屈菜红碱可以有效抑制耐药路邓葡萄球菌的生长,为路邓葡萄球菌抑制剂的研究开发和应用提供新的思路和来源,在医药等领域具有广泛的应用价值。
进一步的,本发明明确了白屈菜红碱对耐药路邓葡萄球菌的抑制作用,将其作为抗生素替代品,能够有效缓解或解决路邓葡萄球菌的耐药及感染问题,降低病死率。
具体实施方式
下面结合实施例对本发明做进一步详细说明。所述实施例仅用于解释本发明,并非对本发明保护范围的限制。
1、人源路邓葡萄球菌的药敏实验
本发明以多株人源路邓葡萄球菌(菌株样本取自宁波市疾病预防控制中心)为出发菌株,选用青霉素、克林霉素、四环素、庆大霉素、利奈唑胺、莫西沙星6种常用抗生素的药敏试纸片进行药敏试验。
挑取培养18~24h的纯菌落均匀溶解于2~5mL无菌生理盐水中,调节其浊度与0.5麦氏比浊管等浊。无菌吸取50~200μL菌液,均匀涂布平板,室温下放置3~5min。用无菌镊子按要求将药敏试纸片(直径为6mm)紧贴于平板表面,每种药敏试纸片三个平行。将接种好的平板放置于36~37℃孵育16~24h。用0.02mm精度的游标卡尺测定抑菌圈直径,以抑菌圈的直径来判断药物的敏感性,抑菌结果根据美国临床试验室国家标准化管理委员会(CLSl2017)标准判定,判定标准见表1,实验结果见表2。结果显示,5#人源路邓葡萄球菌可以耐受3种临床上常见的抗生素,因此将其作为下一步实验的研究对象。
表1.美国临床试验室国家标准化管理委员会(CLSl2017)标准判定结果
表2.人源路邓葡萄球菌的药敏实验中抑菌圈直径结果
注:抑菌圈直径的单位为mm,数值均为三个平行的平均值
2、白屈菜红碱对耐药路邓葡萄球菌的抑制作用
为了充分考虑用药安全,本发明以白屈菜红碱这一单一活性成分为研究对象,以ATCC700328为参考菌株(参考菌株对庆大霉素、利奈唑胺、万古霉素和莫西沙星高度敏感),以5#菌株为实验菌株,进行耐药性抑制作用的研究。挑取培养18~24h的纯菌落均匀溶解于2~5mL LB液体培养基中,调节其浊度与0.5麦氏比浊管等浊,并用酶标仪测定其OD600值。用二甲基亚砜配制浓度为20mg/mL的白屈菜红碱作为药液,利用试管二倍稀释法将药液、菌液及LB液体培养基加入96孔培养板中过夜培养,不同浓度的药液组别均为三个平行,确保实验数据的可信度。
利用酶标仪测定白屈菜红碱对路邓葡萄球菌的最小抑菌浓度(MIC),并以MIC浓度为参照,对进行培养的菌液增加药物稀释梯度,将相应培养液转接于无菌的TSA固体培养基中培养24h,若无单菌落生成,此浓度为白屈菜红碱对路邓葡萄球菌的最低杀菌浓度(MBC),实验结果见表3。
表3.白屈菜红碱对耐药路邓葡萄球菌的抑制结果
由表3可知,白屈菜红碱对耐青霉素、克林霉素及四环素的路邓葡萄球菌的抑制作用较好,其MIC为7.8μg/mL,MBC为16μg/mL。结果表明白屈菜红碱不仅对路邓葡萄球菌参考菌株ATCC700328有抑菌作用,对本发明中的耐药路邓葡萄球菌(例如,5#菌株)也有抑菌作用。
根据以上实验结果,结合中草药来源广泛、不良反应少及不易产生耐药性等特点,可通过获得活性单品成分白屈菜红碱,直接针对临床主要的耐药路邓葡萄球菌发挥抑制作用,可有效缓解或解决路邓葡萄球菌的感染问题,降低病死率,为研究临床分离的路邓葡萄球菌的抑菌剂提供科学依据,为开发抑制耐药路邓葡萄球菌的药物和抗生素替代品提供新的思路和来源。
Claims (10)
1.白屈菜红碱在抑制路邓葡萄球菌生长中的应用。
2.根据权利要求1所述的应用,其特征在于:所述路邓葡萄球菌为人源路邓葡萄球菌。
3.根据权利要求1所述的应用,其特征在于:所述路邓葡萄球菌的耐药性是通过纸片扩散法确定的。
4.根据权利要求1所述的应用,其特征在于:所述路邓葡萄球菌耐受的抗生素选自青霉素、克林霉素、四环素中的一种以上。
5.根据权利要求1所述的应用,其特征在于:所述路邓葡萄球菌选自耐青霉素、克林霉素和四环素的路邓葡萄球菌。
6.根据权利要求5所述的应用,其特征在于:所述白屈菜红碱对路邓葡萄球菌的最低杀菌浓度为16μg/mL,最小抑菌浓度为7.8μg/mL。
7.根据权利要求1所述的应用,其特征在于:所述白屈菜红碱对路邓葡萄球菌具有体外杀灭作用,以及抑制路邓葡萄球菌的体外生长。
8.白屈菜红碱在制备抗路邓葡萄球菌的药物中的应用。
9.根据权利要求8所述的应用,其特征在于:所述路邓葡萄球菌为人源路邓葡萄球菌。
10.根据权利要求8所述的应用,其特征在于:所述路邓葡萄球菌选自对青霉素、克林霉素、四环素中一种以上的抗生素耐药的路邓葡萄球菌。
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