CN108079026A - 一种桑黄提取物在制备抗菌药物中的应用 - Google Patents
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Abstract
本发明公开了一种桑黄提取物在制备抗菌药物中的应用，一种桑黄提取物在制备抗菌药物中的应用，所述桑黄提取物是从干燥桑黄样品中提取得到。本发明探明了桑黄提取物的抗菌性能，桑黄提取物具有良好的抗菌活性，尤其对革兰氏阳性菌更敏感，能够制备抗菌药物提供较大的帮助。
Description
技术领域
本发明涉及一种桑黄提取物，尤其涉及的是一种桑黄提取物在制备抗菌药物中的应用。
背景技术
桑黄是一种珍稀的药用真菌，主要用于止泻、崩漏带下、脾虚泄泻，此外还能利五脏、排毒、活血。
桑黄多糖能有效抑制癌细胞生长和防止其转移是通过细胞调节和体液免疫而起作用的，并且对正常成纤维细胞没有细胞毒作用。桑黄具有明显的抗肝纤维化作用和抗氧化作用。
在过去的30年里，其20多种药效逐渐为人们所探知，如增强机体免疫、保肝护肝、治疗关节炎等(张维博，王家国，李正阔，杨丽群，秦俭，向仲怀，崔红娟。药用真菌桑黄的研究进展，中国中药杂志，2014；39(15)：2838-2845)；桑黄的乙醇提取物已被证实具有消炎和止痛功效。
发明内容
本发明的目的在于克服现有技术的不足，提供了一种桑黄提取物在制备抗菌药物中的应用，能够进一步研究桑黄提取物的抗菌特性。
本发明是通过以下技术方案实现的，本发明的所述桑黄提取物是从干燥桑黄子实体中提取得到的，其具有抗菌活性的提取物是从石油醚和乙酸乙酯萃取部位得到，其主要成分是甾体和酚性成分，本品按干燥品计算，含总酚1.85～3.00％，甾体成分中主要成分是麦角甾醇，本品按干燥品计算，含麦角甾醇0.086～0.120％。
所述提取过程如下：将干燥的桑黄子实体粉碎过16目筛，用95％乙醇溶液提取四次，合并滤液过滤，将滤液浓缩成浸膏，用蒸馏水悬浮，然后用石油醚、乙酸乙酯萃取，将萃取液合并，滤液用旋转蒸发仪蒸干，再用10％DMSO溶液使其溶解并用灭菌蒸馏水采用二倍稀释法稀释。
所述抗菌药物的菌种为表皮葡萄球菌(Staphy lococcus epidermidis)、金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)、大肠杆菌(Escherichia coli)和绿脓杆菌(Pseudomonas aeruginosa)、白色念珠菌(Candidaalbicans)和黑曲霉(Aspergillus nigar)。
本发明相比现有技术具有以下优点：本发明探明了桑黄提取物的抗菌性能，桑黄提取物具有良好的抗菌活性，尤其对革兰氏阳性菌更敏感，能够为制备抗菌药物提供较大的发展思路。
附图说明
图1是桑黄提取物的抑菌直径示意图。
具体实施方式
下面对本发明的实施例作详细说明，本实施例在以本发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。
本实施例首先制备桑黄提取物：
其具有抗菌活性的提取物是从石油醚和乙酸乙酯萃取部位得到，其主要成分是甾体和酚性成分，本品按干燥品计算，含总酚2.7％，甾体成分中主要成分是麦角甾醇，本品按干燥品计算，含麦角甾醇0.107％。
将干燥的桑黄子实体20g粉碎过16目筛，用95％乙醇溶液提取四次，合并滤液过滤，将滤液浓缩成浸膏称重3.2g，用蒸馏水悬浮，后用石油醚、乙酸乙酯萃取，将萃取液合并，滤液用旋转蒸发仪蒸干，称重1.72g，然后用10％DMSO溶液使其溶解并用灭菌蒸馏水采用二倍稀释法稀释。
本实施例选用表皮葡萄球菌(Staphy lococcus epidermidis)、金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)、大肠杆菌(Escherichiacoli)、绿脓杆菌(Pseudomonas aeruginosa)、白色念珠菌(Candida albicans)和黑曲霉(Aspergillus nigar)，上述菌种均购自于黑龙江省微生物研究所。前三种为革兰氏阳性菌，大肠杆菌和绿脓杆菌为革兰氏阴性菌，后两种为真菌。除黑曲霉用察氏培养基于28℃培养5d外，其余菌种均于营养琼脂培养基37℃培养24h。
将上述细菌接种于LB液体培养基中培养活化后，以无菌水制成含菌数约为1×105cfu﹒ml-1菌悬液。将上述真菌菌种挑取一小块接种于PDA平板中央中培养活化。
桑黄提取物抑菌直径的测定：
采用琼脂扩散法用无菌棉签蘸取105cfu﹒mL-1的试验菌悬液，在营养琼脂平板表面均匀涂抹3次，然后绕平板边缘涂抹一周，盖好平皿，室温干燥5min，滤纸片(直径6.0mm)平放于无菌平皿内160℃干热灭菌2h，每片滴加桑黄提取液(200mg﹒mL-1)20μL，开盖室温自然晾干，用无菌镊子取样片轻轻贴放于平板表面，每个平板贴3片试验样片，1片无菌生理盐水对照片。各实验菌株在相应的培养条件下培养。测量抑菌直径，实验重复3次。测量结果如图1所示。由图1可以看出桑黄提取物对革兰氏阳性菌比对革兰氏阴性菌敏感，对真菌则不敏感。10％DMSO无抑菌活性。由此可见，桑黄提取物具有抑菌活性。
采用微量肉汤稀释法测定MIC及MBC：
MIC(minimal inhibitory concentration)为最小抑菌浓度，即药物与一定浓度的菌液作用后，能够抑制可见菌生长的最低浓度，MBC(minimal bactericidalconcentration)为最小杀菌浓度，即在MIC基础上，从每管吸取10μL溶液，点于固体培养基上，继续按MIC培养条件下培养，以杀灭99.9％细菌数量的最低浓度为最小杀菌浓度。采用二倍稀释法将药液用无菌生理盐水稀释系列浓度(0.064～65.8mg/mL)，在96孔板上每孔加100μL不同浓度的药液和100μL菌液，使最终菌液浓度为1×105cfu﹒ml-1，以无菌生理盐水加菌液作为阳性对照，每个实验重复三次。结果如表1所示：
表1 桑黄提取物的MIC和MBC
由表1可见，桑黄提取物对三种革兰氏阳性菌(金黄色葡萄球菌、表皮葡萄球菌、枯草芽孢杆菌)比对革兰氏阴性菌(大肠杆菌和绿脓杆菌)及真菌(白色念珠菌及黑曲霉)的抗菌活性强。通过本研究可以得出，桑黄提取物具有良好的抗菌活性，尤其对革兰氏阳性菌更敏感。
使用本实施例的桑黄提取物制得的桑黄抑菌软膏剂可用于慢性溃疡创伤面，桑黄提取物联合促透剂制成凝胶可用于皮肤瘙痒、湿疹、痤疮，桑黄创伤膏可提高创面修复质量，缩短创面愈合时间，用于皮肤创伤。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
Claims (6)
1.一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述桑黄提取物是从干燥桑黄子实体中提取得到的，其具有抗菌活性的提取物是从石油醚和乙酸乙酯萃取部位得到，其主要成分是甾体和酚性成分，本品按干燥品计算，含总酚1.85～3.00％，甾体成分中主要成分是麦角甾醇，本品按干燥品计算，含麦角甾醇0.086～0.120％。
2.根据权利要求1所述的一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述提取过程如下：将干燥的桑黄子实体粉碎过16目筛，用95％乙醇溶液提取四次，合并滤液过滤，将滤液浓缩成浸膏，用蒸馏水悬浮，然后用石油醚、乙酸乙酯萃取，将萃取液合并，滤液用旋转蒸发仪蒸干，再用10％DMSO溶液使其溶解并用灭菌蒸馏水采用二倍稀释法稀释。
3.根据权利要求1所述的一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述抗菌药物的菌种为表皮葡萄球菌(Staphy lococcus epidermidis)、金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)、大肠杆菌(Escherichiacoli)和绿脓杆菌(Pseudomonas aeruginosa)、白色念珠菌(Candida albicans)和黑曲霉(Aspergillus nigar)。
4.根据权利要求1所述的一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述抗菌药物为用于慢性溃疡创伤面的抑菌软膏剂。
5.根据权利要求1所述的一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述抗菌药物为用于皮肤瘙痒、湿疹、痤疮的凝胶。
6.根据权利要求1所述的一种桑黄提取物在制备抗菌药物中的应用，其特征在于，所述抗菌药物为用于皮肤创伤的创伤膏。
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