CN113476486A - 桑黄的应用以及一种缓解口腔粘膜下纤维化的产品 - Google Patents
桑黄的应用以及一种缓解口腔粘膜下纤维化的产品 Download PDFInfo
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Abstract
本发明涉及医药技术领域，公开了桑黄的应用以及一种缓解口腔粘膜下纤维化的产品。本发明提供了桑黄及其提取物在治疗OSF领域的新应用。RT‑PCR分析结果以及masson纤维化染色结果均显示，对于使用桑黄提取物进行干预的动物模型，各生化指标和纤维化病理特征出现明显改善，这为控制OSF病程进展和治疗OSF提供了更加丰富的治疗途径，也为相关产品的研发提供了充足的基础。
Description
技术领域
本发明涉及医药技术领域，具体涉及桑黄的应用以及一种缓解口腔粘膜下纤维化的产品。
背景技术
嚼食槟榔时，槟榔的木质纤维，以及槟榔产品中的成分，如槟榔碱、石灰、铜离子、亚硝胺类等会刺激口腔黏膜，引起破损，长期刺激可能导致口腔黏膜下纤维化(OSF)。
Kondaiah P等(Molecular pathways regulated by areca nut in theetiopathogenesis of oral submucous fibrosis)在研究结果中指出，OSF的发生主要是因为口腔黏膜在槟榔刺激后TGF-β1表达过度增加，并诱导下游的α-SMA、γ-SMA，Collagentype VI等纤维蛋白成分过量表达并沉积在口腔黏膜，同时MMP3、MMP9表达升高，TIMP1表达降低，抑制了对胶原蛋白纤维的降解，最终导致口腔黏膜处胶原蛋白纤维沉积，引起纤维化病变。
针对OSF目前尚无好的预防和治疗方法，因此对于相关药物的研发具有重要意义。
发明内容
有鉴于此，本发明的目的在于提供桑黄在制备治疗OSF的产品中的新应用；
本发明的另外一个目的在于提供桑黄提取物在制备治疗OSF的产品中的新应用；
本发明的另外一个目的在于提供一种基于桑黄提取物的缓解OSF的有效性产品。
在本发明具体实施方式中，通过对桑黄子实体进行提取多糖，作用于口腔黏膜下纤维化大鼠模型进行干预，结果显示，相对于未实施桑黄多糖的模型组，干预组显著抑制了TGF-β1、α-SMA、γ-SMA，Collagen type VI、MMP9表达，降低了MMP3表达，并促进了TIMP1表达；
同时，大鼠口腔粘膜masson纤维化染色结果显示，模型组中可见大量胶原纤维在粘膜下沉积，表现出典型的纤维化特征；而干预组在施用了桑黄多糖后，纤维蛋白沉积情况出现显著改善；上述实验结果表明桑黄多糖可有效的改善口腔黏膜下纤维化状况，可以作为治疗OSF的活性成分。
基于此，本发明提出了桑黄在制备治疗或缓解OSF的产品中的应用，所述产品包括但不限于药物、功能性食品和保健食品，用于治疗、改善或辅助治疗OSF；进一步地是桑黄作为唯一治疗或缓解OSF的活性成分在制备治疗或缓解OSF的产品中的应用；作为优选，桑黄可以是其提取物，如水提取物或醇提物等；在本发明具体实施方式中，本发明以桑黄多糖作为其提取物进行OSF治疗或改善效果验证。
依据上述应用，本发明还提供了一种缓解OSF的产品，其以桑黄提取物为治疗OSF的活性成分。作为更加优选的方案，所述产品还可以添加药学或食品上允许添加的辅料，和/或用于治疗其他口腔疾病的、不与桑黄提取物，确切地说是桑黄多糖发生干扰的活性成分。
作为优选，本发明具体提供了一种缓解OSF的凝胶产品，其包括桑黄多糖和用于形成凝胶的食品胶体类物质，如明胶、琼脂糖、天然树胶、甘油树脂、卡拉胶、瓜尔豆胶、海藻酸钠等，这些物质加入沸水溶解后可制成凝胶产品。在本发明具体实施方式中，本发明提供了一种由桑黄多糖、明胶和琼脂糖制备而成的凝胶产品。
由以上技术方案可知，本发明提供了桑黄及其提取物在治疗OSF领域的新应用。RT-PCR分析结果以及masson纤维化染色结果均显示，对于使用桑黄提取物进行干预的动物模型，各生化指标和纤维化病理特征出现明显改善，这为控制OSF病程进展和治疗OSF提供了更加丰富的治疗途径，也为相关产品的研发提供了充足的基础。
附图说明
图1所示为模型组和干预组的大鼠口腔黏膜白斑样本RT-PCR分析结果；*、**和***分别表示p<0.001,p<0.01和p<0.05，MMP3结果p＝0.08；
图2所示为模型组和干预组的大鼠口腔粘膜masson纤维化染色结果。
具体实施方式
本发明公开了桑黄的应用以及一种缓解口腔粘膜下纤维化的产品，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明所述应用和产品已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述应用和产品进行改动或适当变更与组合，来实现和应用本发明技术。
本发明实施例中桑黄多糖基于常规的水提醇沉法制备，也可以通过市售途径进行购买，本发明提取桑黄多糖的方法如下：
桑黄子实体水提后滤液浓缩，然后加乙醇沉淀，通过木瓜蛋白酶法脱蛋白，透析、冷冻干燥得到桑黄子实体粗多糖。然后经过阴离子交换树脂分离和凝胶柱纯化得到桑黄多糖样品。
本发明中使用槟榔提取物对SD大鼠造模，槟榔提取物以槟榔为原料，在由氯仿、甲醇和水组成的介质中研磨，离心获取上清液，过滤后滤液冻干，获得槟榔提取物，使用时按10mg/ml溶解重悬于去离子水。
本发明实施例中实验动物为6周龄雄性SD大鼠，每组10只，平均体重200-220g，来源为同一批SD大鼠。
以下就本发明所提供的桑黄的应用以及一种缓解口腔粘膜下纤维化的产品做进一步说明。
实施例1：桑黄多糖的提取
桑黄子实体60℃烘干后粉碎过筛，置于超声波-微波协同萃取仪进行水提，抽滤后合并滤液浓缩，用体积分数80％乙醇醇沉。通过木瓜蛋白酶法脱蛋白，酶解温度为60℃，静置10min后沸水浴灭酶5min，350r/min离心15min，透析、冷冻干燥得到桑黄子实体粗多糖。然后经过DEAE-52阴离子交换树脂分离和Sepharose CL-6B凝胶柱纯化得到桑黄多糖样品。
实施例2：RT-PCR分析和masson纤维化染色
1、造模
槟榔提取物制备方法：将原果产地于海南、生产加工于湖南的槟榔成品，称重30g槟榔在4℃，氯仿：甲醇：去离子水12:5:3环境中机械研磨1小时。随后将提取物在4℃下12000rpm离心10分钟，获取上清液，经0.22um滤器过滤后，于-80℃冷冻干燥机浓缩冻干，获取干燥粉末状的提取物，按10mg/ml溶解重悬于去离子水备用。
通过给予6周龄雄性SD大鼠口腔黏膜下每天注射100ul槟榔产品提取物(10mg/ml)1次，持续30天的方法，建立口腔黏膜下纤维化动物模型，为模型组，使用桑黄多糖干预的为干预组，每组10只SD大鼠。
2、RT-PCR分析
造模过程中，每天通过喂食一次桑黄多糖(100mg/kg体重)的方式，进行干预(干预组)，模型组和干预组其他饲养条件相同。模型组和干预组注射槟榔产品提取物30天后取口腔黏膜白斑样本提取RNA进行RT-PCR分析，结果见图1。
图1显示，桑黄多糖干预显著抑制了槟榔提取物引起的TGF-β1、α-SMA、γ-SMA，Collagen type VI、MMP9表达，同时降低了MMP3表达，并促进了TIMP1表达，表明桑黄多糖可有效的缓解槟榔引起的口腔黏膜下纤维化。
3、masson纤维化染色
模型组和干预组注射槟榔产品提取物30天后，取大鼠口腔粘膜进行masson纤维化染色，结果见图2。
图2模型组结果中蓝色部分即为胶原蛋白纤维，可见大量胶原纤维在粘膜下沉积，表现出典型的纤维化特征；而干预组中纤维蛋白沉积情况出现显著改善。
实施例3：桑黄多糖凝胶产品的制备
1-5g桑黄多糖，2-10g明胶，2-10g中性琼脂糖，50ml沸水溶解后制成凝胶，4度低温保存。其中，各原料的质量与水的体积可以成倍调整。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.桑黄在制备治疗口腔黏膜下纤维化的产品中的应用。
2.桑黄提取物在制备治疗口腔黏膜下纤维化的产品中的应用。
3.根据权利要求2所述应用，其特征在于，所述桑黄提取物为水提醇沉提取物。
4.根据权利要求2或3所述应用，其特征在于，所述桑黄提取物为桑黄多糖。
5.一种缓解口腔黏膜下纤维化的产品，其特征在于，以桑黄提取物为治疗口腔黏膜下纤维化的活性成分。
6.根据权利要求5所述产品，其特征在于，还包括药学或食品上可接受的辅料和/或不与桑黄提取物发生干扰的治疗其他口腔疾病的活性成分。
7.根据权利要求6所述药物，其特征在于，所述药用辅料为食品胶类物质。
8.根据权利要求7所述产品，其特征在于，所述食品胶类物质选自明胶、琼脂糖、天然树胶、甘油树脂、卡拉胶、瓜尔豆胶和海藻酸钠。
9.根据权利要求5所述产品，其特征在于，所述产品为凝胶产品。
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