CN117771231A - 一种山竹提取物中呫吨酮类化合物在防治奶牛乳腺炎致病菌中的用途 - Google Patents
一种山竹提取物中呫吨酮类化合物在防治奶牛乳腺炎致病菌中的用途 Download PDFInfo
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Abstract
本发明公开一种山竹提取物中呫吨酮类化合物在防治奶牛乳腺炎致病菌方面的用途,属于天然药物化学领域。该类化合物对奶牛乳腺炎致病菌无乳链球菌Streptococcus agalactiae ATCC12386,无乳链球菌Streptococcus agalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiae ATCC27957,大肠杆菌Escherichia coli ATCC 25922和金黄色葡萄球菌Staphylococcus aureus Newman表现出优异的抑制活性。本发明所涉及化合物绿色高效且资源丰富,在代替抗生素成为防治奶牛乳腺炎天然产物来源化合物方面极具应用价值。
Description
技术领域
本发明属于天然药物化学领域,公开了从一种山竹提取物中呫吨酮类化合物在防治由无乳链球菌Streptococcus agalactiae ATCC12386,无乳链球菌Streptococcusagalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiae ATCC27957,大肠杆菌Escherichia coli ATCC 25922和金黄色葡萄球菌Staphylococcus aureus Newman引起的奶牛乳腺炎中的用途。
背景技术
奶牛乳腺炎作为奶牛养殖业最常见的疾病之一,可引起发病奶牛泌乳量减少,影响牛乳品质和营养含量,甚至还能导致奶牛生产性能的丧失,使成年奶牛过早淘汰。导致奶牛乳腺炎发生的因素很多,包括物理性刺激、化学物质侵染以及病原微生物的入侵,然而近50%的奶牛乳腺炎是由细菌感染引起的。奶牛乳腺炎给全世界的奶牛养殖产业和乳制品加工业造成了巨大的经济损失,也是影响奶牛健康和乳制品安全的关键因素。此外,由此带来的食品安全问题严重威胁着人类健康。
目前,应用抗生素仍是奶牛乳腺炎的主要治疗策略。然而,反复和不当使用抗生素可诱导菌株对抗生素产生耐药性,严重影响疗效。同时,抗生素残留的发生,会导致牛奶中蓄积药物,还会对奶牛造成肠道菌群失调,对水和土壤环境造成负面影响,也将危害人们的生活环境以及日常饮食健康。乳腺炎病原体的抗生素耐药性已成为全球广泛关注的问题,这迫使人们开发安全有效的“替抗”的活性化合物来维持奶牛乳房的健康。
植物资源是活性天然产物,其产生次生代谢产物超过400,000种,且其中多数都具有良好的抗菌活性。天然产物一直是药物研发中的重要来源。在过去的二十几年中,超过1/3的FDA批准上市药物为天然产物及其衍生物。相比于化学合成化合物,天然产物具有一些独特的性质,比如结构复杂度更高,骨架更加新颖,类药性更好等。因此,天然产物是一个值得关注的宝库,对于药物获得有巨大潜力。
α-倒捻子素是从植物山竹中提取的一种呫吨酮类化合物。山竹属于藤黄科、藤黄属常绿乔木山竹的果实,在泰国山竹一直作为传统医药用于治疗腹痛、腹泻、痢疾等疾病。山竹果壳的主要代谢产物为氧化蒽醌,其中α-倒捻子素最为经典,它含量丰富,具有显著的抗炎、抗氧化和抗肿瘤的作用,其活性高、低毒、副作用小等。本发明使用由山竹中提取分离出的S1-S4化合物(其中S1是由山竹提取物分离得到的α-倒捻子素),评价其对奶牛乳腺炎的主要致病菌,包括无乳链球菌Streptococcus agalactiae ATCC12386,无乳链球菌Streptococcus agalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiaeATCC27957,大肠杆菌Escherichia coli ATCC 25922和金黄色葡萄球菌Staphylococcusaureus Newman的抗菌活性,显示出优异的抑制活性。因此,在治疗奶牛乳腺炎方面具有潜在的应用价值,值得进一步研究和开发。
发明内容
本发明的目的是提供由山竹提取物分离的呫吨酮类化合物在治疗奶牛乳腺炎方面的新用途,用于防治由无乳链球菌、停乳链球菌、大肠杆菌和金黄色葡萄球菌引起的奶牛乳腺炎。
为了实现以上目的,本发明提供了如下技术方法:
本发明提供了一种在抗奶牛乳腺炎致病菌中的新用途,其对无乳链球菌Streptococcus agalactiae ATCC12386,无乳链球菌Streptococcus agalactiaeATCC27956,停乳链球菌Streptococcus dysgalactiae ATCC27957,大肠杆菌Escherichiacoli ATCC 25922和金黄色葡萄球菌Staphylococcus aureus Newman的给药浓度为800、400、200、100、50、25、12.5、6.25、3.12μg/mL等。本发明涉及的化学结构式如下:
本发明提供的抗菌剂具有以下优势:
1)本发明发现对奶牛乳腺炎致病菌表现出优异的抑制作用,在治疗奶牛乳腺炎方面极具应用价值。
2)天然源抗菌剂因其具有低毒、低残留及不易产生抗性等优点,在防治奶牛乳腺炎方面具有无法替代的独特性。在我国分布广泛、资源丰富,具有绿色高效、不易产生耐药性等特点,在防治奶牛乳腺炎方面具有应用价值,值得进一步研究和开发。
具体实施方式
为了更好地理解本发明,通过以下具体实施例对本发明的上述内容做进一步的详细说明。但不应将此理解为对本发明的限制。以下是本发明的具体实施例,对本发明的技术方案做进一步描述,但本发明的内容不仅仅局限于实施例所述的范围,凡是不背离本发明构思的改变或等同替代均包括在本发明的保护范围之内。
实施例1:呫吨酮类化合物抗奶牛乳腺炎致病菌活性测定
1)供试药剂:α-倒捻子素及S1-S4化合物。
2)供试菌种:无乳链球菌Streptococcus agalactiae ATCC12386,无乳链球菌Streptococcus agalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiaeATCC27957,大肠杆菌Escherichia coli ATCC 25922和金黄色葡萄球菌Staphylococcusaureus Newman。
3)测试方法:
本实验中所用的菌株为实验室-80℃含30%甘油冻存的菌株。将无乳链球菌和停乳链球菌冻存菌株取出,分别在TSA(胰酪大豆胨琼脂培养基)+5%羊血琼脂平板上面进行划线,在37℃下恒温培养直到长出单菌落。分别挑取单菌落至脑心浸出液肉汤(BHI)液体培养基(胰蛋白胨:10.0g,牛心浸粉:17.5g,氯化钠:5.0g,葡萄糖:2.0g,磷酸氢二钠(12H2O):2.5g,蒸馏水:1L;121℃灭菌20min),在37℃、140rpm恒温摇床振荡培养到对数生长期。同样的,将大肠杆菌和金黄色葡萄球菌的冻存菌株取出,分别在MH固体培养基上面进行划线,在37℃下恒温培养直到长出单菌落。分别挑取单菌落至MH肉汤培养基(牛肉粉:2.0g,可溶性淀粉:1.5g,酸水解酪蛋白:17.5g,蒸馏水:1L;121℃灭菌20min)液体培养基。然后,分别将处于对数生长期的菌株用相应的液体培养基稀释至约106CFU/mL备用。将化合物分别用DMSO溶解,加入液体培养基中,混合均匀,将化合物配制成浓度为1600μg/mL的含药液体培养基。取50μL含药培养基和相同体积的含约106CFU/mL细菌培养物加入到96孔板的孔中,最终化合物浓度都为800μg/mL。含等量DMSO的相同浓度100μL菌液做对照。将96孔板在37℃恒温培养箱中培养12-24h直至对照组菌液长出,在酶标仪上测定孔中菌液的OD值(OD600)。并且另外测定100μL液体培养基和浓度为800μg/mL药剂的OD值,对培养基和药剂本身造成的OD值进行矫正。校正OD值和抑制率的计算公式如下:
校正OD值=含菌培养基OD值-无菌培养物OD值;
抑制率=(校正后对照培养基菌液OD值-校正后含药培养基OD值)/校正后对照培养基菌液OD值×100%
将活性化合物的含药液体培养基在96孔板中通过二倍稀释法稀释得到系列浓度的50μL含药培养基,然后根据实施例1中的试验方法测定系列浓度对应的抑制率。将抑制率大于90%的最低浓度定义为MIC,测定得到的活性数据见表1。
表1.化合物抗奶牛乳腺炎致病细菌的MIC值
由实验结果可知,本发明涉及的化合物对测试菌株均表现出优异的杀菌活性,其中由山竹中分离得到的α-倒捻子素S1与所购买的α-倒捻子素活性相当,其MIC值在1.56~6.25μg/mL,表现出显著的抗菌活性。
综上所述,本发明所述的化合物对无乳链球菌Streptococcus agalactiaeATCC12386,无乳链球菌Streptococcus agalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiae ATCC27957,大肠杆菌ATCC 25922,金黄色葡萄球菌Newman表现出显著的抗菌活性,可在防治奶牛乳腺炎方面进一步研究和开发。
Claims (3)
1.本发明涉及一种呫吨酮类化合物在防治奶牛乳腺炎中的用途。
2.根据权利要求1所述的用途,其中,所涉及化合物结构式如下:
3.根据权利要求1和2所述的呫吨酮化合物在防治由无乳链球菌Streptococcusagalactiae ATCC12386,无乳链球菌Streptococcus agalactiae ATCC27956,停乳链球菌Streptococcus dysgalactiae ATCC27957,大肠杆菌ATCC 25922,金黄色葡萄球菌Newman引起的奶牛乳腺炎中的用途。
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