CN105920057A - 一种淫羊藿苷的用途 - Google Patents
一种淫羊藿苷的用途 Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
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Abstract
本发明公开了一种淫羊藿苷的用途，该淫羊藿苷可用于制备保健食品或食物添加剂。一种应用淫羊藿苷的防止动脉硬化的药物组合物，该药物组合物由以下成分及其质量百分比组成：淫羊藿苷5~15%、银杏黄酮1~12%、洛伐他汀1~10、吉非贝齐5~10%、胆汁酸5~10%、填充剂43~83%。
Description
技术领域
本发明涉及天然药物领域，具体涉及一种淫羊藿苷的用途。
背景技术
淫羊藿苷（Icariin）是从小檗科植物淫羊藿（Epimedium brevicornum Maxim）全草中提取的活性成分，属黄酮类化合物。淫羊藿苷具有抗炎、抗肿瘤、免疫调节和抗氧化作用。
氧化低密度脂蛋白（ox-LDL）可诱导血管平滑肌细胞（VSMC）损伤，而受损的VSMC由中膜向内膜下迁移、增殖、合成和分泌大量细胞外基质，PCNA即是VSMC增殖的重要标志物之一，其表达水平的高低可反映细胞的增殖活性。
丝裂原活化蛋白激酶（MAPK）是细胞内促增殖和传递应激信号的关键激酶，该家族包括ERK、JNK、p38等亚族与细胞增殖分化相关的信号通路关系密切，该信号转导通路在未受刺激时，处于静止状态；在受到生长因子的刺激时，在上游激酶的作用下，发生级联磷酸化而激活，MAPK被激活后进入细胞核，使一些转录因子磷酸化，改变细胞基因的表达状态，参与细胞的生长、分化、增殖等多种生理过程的调控，在动脉粥样硬化的起病和发展过程中起着极其重要的作用。
因此，基于已知的ＶＳＭＣ增殖调节机制，寻找药物干预与此相关的病理过程已经成为目前防治心血管疾病的重要策略之一，如“羟基红花黄色素A通过影响PCNA表达和MEK-ERK1/2信号通路抑制大鼠血管平滑肌细胞增殖”（《中国药理学通报》2015年滴7期）。
发明内容
本发明为解决上述技术问题提供一种淫羊藿苷一种淫羊藿苷的用途。
本发明为解决上述技术问题所采取的方案为：一种淫羊藿苷的用途，其特征在于：
该淫羊藿苷可用于制备保健食品或食物添加剂。
一种应用淫羊藿苷的防止动脉硬化的药物组合物，该药物组合物由以下成分及其质量百分比组成：淫羊藿苷5~15%、银杏黄酮1~12%、洛伐他汀1~10、吉非贝齐5~10%、胆汁酸5~10%、填充剂43~83%。
作为优选，该药物组合物由以下成分及其质量百分比组成：淫羊藿苷13%、银杏黄酮10%、洛伐他汀7%、吉非贝齐9%、胆汁酸10%、填充剂70%。
或者，该药物组合物由以下成分及其质量百分比组成：淫羊藿苷10%、银杏黄酮7%、洛伐他汀10%、吉非贝齐8%、胆汁酸10%、填充剂75%。
附图说明：
图1 淫羊藿苷可剂量依赖性抑制ox-LDL诱导的VSMC增殖
图2 MTT实验结果
图3 细胞周期测定实验结果
图4 淫羊藿苷可剂量依赖性的降低ox-LDL诱导增殖的VSMC的PCNA蛋白表达水平
图5 淫羊藿苷可剂量依赖性的降低ox-LDL诱导增殖的VSMC的ERK1/2蛋白磷酸化。
具体实施方式
以下结合附图和实施例作进一步详细描述：
实施例1：MTT实验
实验方法：
将处于对数生长期间的VSMC接种于96孔板中，24h后给予ox-LDL(50 μg/mL)刺激，刺激24h后给予不同剂量的淫羊藿苷(0，10，20，40 μM)。24h、48h后每孔中加入20μL(5mg/mL)MTT，置培养箱中，温度37°C ，孵育4h，弃去MTT溶液，加入DMSO(150μL/孔)溶解结晶，15min后取出置于酶标仪，于490nm波长处测定其吸光度（OD值）。给药组细胞存活率用以下公式计算:存活率=（处理组OD值/对照组OD值）*100%，结果如图1、2。
实施例2：Westernblot实验
实验方法：
将处于对数生长期间的VSMC接种于6孔板中，24h后给予ox-LDL(50 μg/mL)刺激，24h后给予不同剂量的淫羊藿苷(0，10，20，40 μM)，置培养箱中，温度37°C ，孵育24h，收集细胞，制成westblot实验样品，经BCA试剂盒蛋白定量后，加5×上样缓冲液煮沸5 min后，即可进行SDS-PAGE。配制10% SDS-PAGE分离胶，***梳齿，每孔中加入变性蛋白20 μg，在Marker加样孔中加入10 μl的Marker。接通电源，稳压120 V，45 min进行电泳，待蛋白样品中的溴酚蓝泳至距分离胶下缘1 cm时，停止电泳。制作三明治，转膜槽内加入预冷的转移缓冲液，恒流180 mA转膜2 h。取出PVDF膜在室温封闭2～3 h。ERK 1/2，p-ERK 1/2，PCNA等蛋白一抗稀释液，与PVDF膜4 ℃摇床孵育过夜。洗液冲洗3次，每次10 min ，辣根酶标记山羊抗兔二抗（1:5000）稀释，常温下与PVDF膜摇床2 h。洗液冲洗3次，每次10 min，按照ECL试剂盒说明书进行操作，将发光液均匀地滴在PVDF膜上，反应1 min，最后将膜放入凝胶成像***检测，结果如图4、5。。
实施例3：细胞周期测试
实验方法：
将处于对数生长期间的VSMC接种于6孔板中，24h后给予ox-LDL(50 μg/mL)刺激，24h后给予不同剂量的淫羊藿苷(0，10，20，40 μM)，置培养箱中，温度37°C ，孵育24h，收集细胞到15ml的管中，离心，10mLPBS洗细胞，沉淀细胞并重悬，共2次，70%冰乙醇固定，加入PCB溶液室温低渗处理后，离心去上清，加入RNaseA消化后，加入PI，上流式细胞仪检测细胞周期，结果如图1、3。
其中，MTT染色及流逝细胞周期结果显示，淫羊藿苷对ox-LDL引起的VSMC增殖具有显著的抑制作用。Westernblot实验结果证实，淫羊藿苷可以剂量依赖性的降低ox-LDL刺激后增殖的VSMC的PCNA蛋白表达水平，并可剂量依赖性的降低ERK1/2蛋白磷酸化。
实施例4
一种应用淫羊藿苷的防止动脉硬化的药物组合物，分别称取淫羊藿苷52g、银杏黄酮35g、洛伐他汀28g、吉非贝齐38g、胆汁酸37g、填充剂110g，混合均匀制成药物1000份，每份0.3g。
对比实验
实验方法：
取两只动脉硬化的大鼠，分别使用淫羊藿苷（对照组）和 淫羊藿苷组合物（实验组）进行喂药，淫羊藿苷组合物由以下成分及其质量百分比组成：淫羊藿苷13%、银杏黄酮10%、洛伐他汀7%、吉非贝齐9%、胆汁酸10%、填充剂70%。并在37℃温度下，孵育24h，通过解剖发现，结果如下表1：
表1 对比实验
| 层状胶原 | 黄斑 | 肉芽组织和纤维化 | 血管壁 | |
| 实验组 | 隐约可见 | 无 | 隐约可见 | 柔软 |
| 对照组 | 明显可见 | 隐约可见 | 明显可见 | 硬化 |
虽然本发明已以实施例公开如上，但其并非用以限定本发明的保护范围，任何熟悉本领域的技术人员，在不脱离本发明的构思和范围内所做的更动和润饰，均应属于本发明所附权利要求的保护范围。
Claims (3)
1.一种淫羊藿苷的用途，其特征在于：该淫羊藿苷可用于制备保健食品或食物添加剂。
2.一种应用淫羊藿苷的防止动脉硬化的药物组合物，其特征在于：该药物组合物由以下成分及其质量百分比组成：淫羊藿苷5~15%、银杏黄酮1~12%、洛伐他汀1~10、吉非贝齐5~10%、胆汁酸5~10%、填充剂43~83%。
3.根据权利要求1所述的药物组合物，其特征在于：该药物组合物由以下成分及其质量百分比组成：淫羊藿苷13%、银杏黄酮10%、洛伐他汀7%、吉非贝齐9%、胆汁酸10%、填充剂70%。
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