CN104587103A - 一种治疗出血性中风中药挥发油提取物成分群的提取方法及其应用 - Google Patents
一种治疗出血性中风中药挥发油提取物成分群的提取方法及其应用 Download PDFInfo
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Abstract
本发明公开了一种治疗出血性中风中药挥发油提取物成分群的提取方法及其应用。大黄、丹皮、赤芍、石菖蒲按一定重量配比，粉碎后加水浸泡，水蒸汽蒸馏法提取挥发油，经GC-MS分析有70种挥发油成分，其中β-细辛醚占30.8%-36.2%、丹皮酚14.8%-18.2%、α-细辛醚3.8%-7.4%、菖蒲酮3.8%-5.2%。该中药挥发油提取物能显著降低自发性高血压雄性大鼠神经元特异性烯醇化酶、Ｓ-100β蛋白、IL-1、TNF-α、血浆纤维蛋白原水平，可用于制备治疗出血性中风药物的用途。
Description
一、技术领域
本发明涉及中医药技术领域，具体地说是一种治疗出血性中风挥发油提取物成分群的提取方法及其应用。
二、背景技术
出血性中风(脑出血)是国内外临床上常见的疾病，该病发病急骤，进展迅速，病死率高，患者预后情况直接影响到病后的生存质量。神经元特异性烯醇化酶(SNE)和S-100β蛋白是判断脑损害严重程度的重要指标，其中，SNE是神经元及神经内分泌细胞的一个标志酶，当脑组织发生缺血缺氧时，脑脊液中SNE水平升高。S-100β蛋白是脑特异性蛋白，血清中S-100β蛋白水平越高，脑损害程度越重。当前西医治疗脑出血的方法为手术治疗和内科综合治疗两大类，有一定危险性且疗效不够显著。
中医认为出血性中风源于血分瘀热，搏击不解，闭阻脑窍，而令邪热愈炽，进而化火生风生痰，造成瘀热阻窍，故治疗时以凉血散瘀、通腑泄热为主。中药大黄清热泻火、丹皮凉血散瘀、赤芍凉血活血、石菖蒲开窍醒神。因此，本发明采用水蒸汽蒸馏法提取制备该中药组合物挥发油提取物，用自发性高血压大鼠模型评价其抗炎、活血及防治脑损伤作用，用于治疗出血性中风药物的研发。
三、发明内容
针对临床出血性中风病死率和致残率高的现状，本发明的目的是提供一种具有治疗出血性中风挥发油提取物成分群的提取方法及其应用。
本发明的目的是通过以下的技术方案来实现的：
1.一种治疗出血性中风挥发油提取物成分群的提取方法，提取步骤如下：
(1)原料:大黄1-5份、丹皮1-5份、赤芍1-5份、石菖蒲1-6份；
(2)将上述原料药粉碎至20-60目，加5-20倍量水浸泡5-12h,水蒸汽蒸馏法提取4-8小时，收集挥发油提取物；
(3)挥发油成分分析：挥发油提取物用GC-MS分析有70种挥发油成分，其中β-细辛醚 占30.8％-36.2％、丹皮酚14.8％-18.2％、α-细辛醚3.8％-7.4％、菖蒲酮3.8％-5.2％。
2.一种挥发油提取物成分群在制备治疗出血性中风药物中的应用
有益效果：
本发明的中药挥发油提取物成分群，主要由β-细辛醚、丹皮酚、α-细辛醚、菖蒲酮等70多个挥发油成分组成，能显著降低自发性高血压大鼠神经元特异性烯醇化酶(SNE)、S-100β蛋白、IL-1、TNF-α、血浆纤维蛋白原水平，可用于制备治疗出血性中风药物的用途。
四、挥发油成分群的药理试验
1.造模及给药
自发性高血压雄性大鼠随机分为4组，每组8只，以Wistar雄性大鼠为空白对照组(生理盐水组)，高血压大鼠分别为模型组(生理盐水组)，阳性对照组(阿司匹林组)、挥发油提取物高、低剂量组。将各组按10mL·kg-1每天灌胃给药1次，连续15天。第15天灌胃给药1h后颈动脉取血5ml于加有0.038g·ml-1枸橼酸钠的离心管中，以3000r·min-1转速离心10min，取上层血浆，Elisa试剂盒测定大鼠S-100B蛋白、NSE、IL-1、TNF-α、血浆纤维蛋白原。
统计采用SPSS13.0统计软件对数据进行双样本t检验及方差分析，实验数据以表示。
2.对S-100B和NSE的影响
实验结果(表1)表明：模型组S-100B和NSE较空白组显著升高(P＜0.01)，与模型组比较，阳性对照组及挥发油不同剂量组可显著降低S-100B和NSE水平(P＜0.01)。
表1挥发油提取物对S-100B和NSE的影响(n＝8)
注：与空白组比##P﹤0.01；与模型组比**P﹤0.01
3.对抗炎因子的影响
实验结果(表2)表明：自发性高血压大鼠模型组IL-1、TNF-α水平较空白组显著升高(P＜0.01)。与模型组比较，阳性对照组及挥发油提取物高、低剂量组能显著降低自发性高血压大鼠IL-1、TNF-α水平(P＜0.01)。
表2挥发油提取物对炎症因子的影响(n＝8)
注：与模型组比较**P﹤0.01；与空白组比较##P﹤0.01
4.对血浆纤维蛋白原的影响
实验结果(表3)表明：自发性高血压雄性大鼠纤维蛋白原水平较空白组显著升高(P＜0.01)。与模型组比较，阳性对照组及挥发油提取物高、低剂量组能显著降低自发性高血压雄性大鼠血浆纤维蛋白原水平(P＜0.01)。
表3对大鼠纤维蛋白原测定结果(n＝8)
注：与模型组比较**P﹤0.01；与空白组比较##P﹤0.01
五、挥发油提取物化学成分GC-MS分析
色谱柱HP-5MS Phenyl Methyl Siloxane石英毛细管色谱柱(0.25mm×30m，0.25μm)；载气为氦气，进样量1μL，流速1μL·mL-1，分流比20∶1,进样口温度250℃，检测器温度250℃；程序升温：初始温度70℃，保持2min，以2℃/min速率升温至150℃，保持10min，以2℃/min的速率升温至180℃，保持20min，以2℃/min的速率升温至230℃，保持5min。质谱分辨率500，电离方式EI，电子能量70eV，离子源温度200℃，加速电压4kV，扫描速度1s·dec-1，扫描范围m/z20～500。
采用GC-MS分析挥发油提取物化学成分(图1)，经数据处理系统对其内存谱库自动检索，人工谱图解析，结合文献资料，鉴定出70个化合物(表4)，占总挥发油量的90.1％-95.4％，其中β-细辛醚占32.1％-38.8％、丹皮酚10.2％-18.2％、α-细辛醚3.5％-7.4％、菖蒲酮2.2％-6.2％。 表4挥发油提取物成分
经研究结果表明，由本发明制备的中药挥发油提取物成分群，由β-细辛醚、丹皮酚、α-细辛醚、菖蒲酮等70多个挥发油成分组成，能显著降低自发性高血压雄性大鼠神经元特异性烯醇化酶、S-100β蛋白、IL-1、TNF-α、血浆纤维蛋白原水平，可用于制备治疗出血性中风药物的用途。
六、附图说明
图1挥发油提取物GC-MS总离子流图。
七.具体实施方式
实施例1：治疗出血性中风挥发油提取物的提取方法，提取步骤如下：
(1)原料药:大黄1份、丹皮1份、赤芍1份、石菖蒲1份；
(2)将上述原料药粉碎至20目，加5倍量水浸泡5h,水蒸汽蒸馏法提取4小时，收集挥发油提取物；
(3)挥发油成分分析：用GC-MS分析挥发油提取物的化学成分，共有70多挥发油成分，其中β-细辛醚占34.8％、丹皮酚14.8％、α-细辛醚5.2％、菖蒲酮3.8％。
实施例2：治疗出血性中风挥发油提取物的提取方法，提取步骤如下：
(1)原料药:大黄3份、丹皮2份、赤芍3份、石菖蒲4份；
(2)将上述原料药粉碎至40目，加10倍量水浸泡10h，水蒸汽蒸馏法提取6小时，收集挥发油提取物；
(3)挥发油成分分析：用GC-MS分析挥发油提取物的化学成分，共有70多挥发油成分，其中β-细辛醚占30.8％、丹皮酚17.8％、α-细辛醚3.8％、菖蒲酮4.8％。
实施例3：治疗出血性中风挥发油提取物的提取方法，提取步骤如下：
(1)原料药:大黄5份、丹皮5份、赤芍5份、石菖蒲6份；
(2)将上述原料药粉碎至60目，加20倍量水浸泡12h,水蒸汽蒸馏法提取8小时，收集挥发油提取物；
(3)挥发油成分分析：用GC-MS分析挥发油提取物的化学成分，共有70多挥发油成分，其中β-细辛醚占36.2％、丹皮酚18.2％、α-细辛醚7.4％、菖蒲酮5.2％。
Claims (2)
1.一种治疗出血性中风的中药挥发油提取物成分群的提取方法，提取步骤如下：
（1）原料药: 大黄1-5份、丹皮1-5份、赤芍1-5份、石菖蒲1-6份；
（2）将上述原料药粉碎至20-60目，加5-20倍量水浸泡5-12h, 水蒸汽蒸馏法提取4-8小时，收集挥发油提取物；
（3）挥发油成分分析：挥发油提取物经GC-MS分析有70种挥发油成分，其中β-细辛醚占30.8%-36.2%、丹皮酚14.8%-18.2%、α-细辛醚3.8%-7.4%、菖蒲酮3.8%-5.2%。
2.如权利要求1所述的中药挥发油提取物成分群在制备治疗出血性中风药物中的应用。
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| CN101721572A (zh) * | 2009-12-30 | 2010-06-09 | 南京中医药大学 | 一种治疗出血性中风的中药复方制剂及其制备方法和应用 |
| WO2013148122A1 (en) * | 2012-03-30 | 2013-10-03 | University Of Central Florida Research Foundation, Inc. | Methods and compositions using fgf-9 to enchance neovascularization and regeneration |
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| CN101721572A (zh) * | 2009-12-30 | 2010-06-09 | 南京中医药大学 | 一种治疗出血性中风的中药复方制剂及其制备方法和应用 |
| WO2013148122A1 (en) * | 2012-03-30 | 2013-10-03 | University Of Central Florida Research Foundation, Inc. | Methods and compositions using fgf-9 to enchance neovascularization and regeneration |
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| 过伟峰等: "凉血通瘀中药治疗出血性中风急性期56例疗效观察", 《中华中医药杂志》 * |
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| WO2023000247A1 (zh) * | 2021-07-22 | 2023-01-26 | 四川大学 | α-细辛脑在制备预防或治疗出血性脑卒中药物中的应用 |
| EP4205734A4 (en) * | 2021-07-22 | 2024-05-01 | Sichuan University | APPLICATION OF ALPHA-ASARONE IN THE PREPARATION OF DRUGS FOR THE PREVENTION OR TREATMENT OF HEMORRHAGIC STROKE |
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