CN114796290B - 香青藤或其提取物在制备抗脑缺血药物中的应用 - Google Patents
香青藤或其提取物在制备抗脑缺血药物中的应用 Download PDFInfo
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Abstract
本发明公开了香青藤或其提取物在制备抗脑缺血药物中的应用，所述香青藤(Illigera aromaticfa S.Z.Huang et S.L.Mo)为莲叶桐科青藤属香青藤的干燥藤茎，对香青藤进行水提得到香青藤的水提物或采用有机溶剂对香青藤进行提取得到香青藤的有机溶剂提取物，提取方法为：取香青藤，加入溶剂进行浸泡处理，再加热进行提取，合并多次提取的药液进行浓缩即得。本发明首次发现香青藤及其提取物具有治疗脑缺血的成药潜力，可用于制备治疗脑缺血的药物，且药物制剂的制备方法简单，成本低，成药经济效益好。
Description
技术领域
本发明属于中医药技术领域，具体涉及一种香青藤或其提取物在制备抗脑缺血药物中的应用。
背景技术
脑卒中包括缺血性脑卒中和出血性脑卒中，缺血性脑卒中约占60%-80%。缺血性脑卒中是由局部脑供血、供氧不足或中断引起的脑部局灶性功能障碍，常见的有短暂脑缺血发作、脑栓塞、脑血栓等，是世界范围内重大的脑血管疾病，是临床常见的为危重病、多发病，往往预后较差，具有发病率和致残致死率高的特点，目前尚缺乏治疗或延缓脑缺血损伤的有效方法。缺血-再灌注损伤(ischemia reperfusion injury，IRI) 是指组织在遭受一定时间缺血，恢复血液供应后，组织损伤程度加重的病理现象。缺血引起的脑组织缺氧以及缺血再灌注，触发了一系列脑缺血级联反应，最终导致神经元死亡和神经功能障碍。
现代医学研究关于脑缺血的发生机制有能量代谢障碍、血小板聚集、血管收缩及微循环障碍、自由基损伤、兴奋性氨基酸毒性作用、细胞内钙离子超载、炎症损伤、NO和NOS的作用、细胞凋亡等，病理生理机制复杂；脑缺血损伤是由于脑部血液循环障碍，造成细胞能量代谢衰竭，从而启动的损伤级联反应的结果。该损伤级联反应主要涉及兴奋性神经毒性、酸中毒、炎症反应、氧化和硝化应激、围梗死去极化、细胞凋亡等方面。现代医学防治脑缺血多采用药物性干预，临床可用于防治脑缺血的药物主要有抗血小板药的阿司匹林、氯吡格雷、噻氯匹定和阿司匹林 - 双嘧达莫，抗凝剂的肝素、低分子肝素，口服抗凝剂的华法林、双香豆素针，溶栓的尿激酶 (UK)、链激酶 (SK)，血管扩张剂麦角溴烟酯，自由基清除剂超氧化物歧化酶、地塞米松、甘露醇、维生素E，钙拮抗剂尼莫地平，谷氨酸受体拮抗剂，NOS抑制剂及他汀类药物等，其中以阿司匹林抗血小板聚集的应用最多。但是现有的药物由于价格较贵且给药方式受限、副作用多而不适宜反复应用或因用药时间长、给药剂量大等使应用受到了限制。
近些年来，国内外学者将目光投向天然药物（中药或中药制剂）的开发，中药治疗脑血管疾病的历史悠久，且中药具有副作用小等优点；中药保护缺血性脑损伤多通过降低兴奋性神经递质和酸的毒性作用、抑制脂质过氧化和硝化反应、减轻炎症反应、促进血管新生和抑制细胞凋亡等途径来实现，具有多靶点、多层次、多环节的脑保护作用，能从多条路径干预脑缺血，减轻脑缺血再灌注的损伤。如，河南中医学院在专利CN 101978987 B、CN102008521 B、CN 103417679 B分别提出了冬凌草提取物、凌霄花提取物、玫瑰花总黄酮在制备治疗抗脑缺血药物中的应用；西北大学在专利CN 103800826 B提出了黄姜提取物在制备抗脑缺血药物中的应用；广西壮族自治区中医药研究院在专利CN 112245456 A提出了一种拟黑多刺蚁提取物在制备抗脑缺血药物中应用。尚远宏和徐晓玉总结了截至2013年近10 年来对脑缺血具有保护作用的中药及其提取物，包括植物药、动物药和矿物药共35 种单味药，国内外文献中的实验研究都表明从中药得到的单体成分(如银杏内酯B、川芎嗪、人参皂苷 Rb3和丹参酮Ⅱ等)或提取物 (如山楂醇提物、当归水提物和地龙提取物等)，及多种药物组方(如银杏达莫注射液、参龙健脑胶囊和参芎注射液等)可通过脑缺血的病理生理机制中的各种途径来实现对缺血性脑损伤的保护作用，充分说明中药及其提取物对于缺血性脑损伤保护作用的研究前景广阔（见尚远宏,徐晓玉.中药及其提取物对脑缺血保护作用的实验研究进展[J]，《中国中药杂志》2013年8期）。
香青藤(Illigera aromaticfa S.Z.Huang et S.L.Mo)为莲叶桐科青藤属香青藤的干燥藤茎，气香，味辛、凉，主要分布在广西，是我国特有的藤本资源。香青藤具有祛风除湿、行气止痛、舒筋活络等功效，用于风湿骨痛、跌打损伤、肥大性脊柱炎等。香青藤常用作民间草药,广西民间常用其茎泡茶饮用,作为镇痛、止痢、解热药物使用。香青藤中含有糖、多糖、苷类、皂苷、有机酸、黄酮类、蒽醌类、香豆素、内酯、植物甾醇、三萜、挥发油、油脂等成分。目前针对香青藤研究较少，主要集中在香青藤的化学成分分析方面，对其化学成分的药理作用报道较少。而在香青藤抗脑缺血方面，目前未见有相关的报道。所以，如何提高香青藤的利用率，使之产生最大的功效，发挥出应有的效能，仍是医学医药研究的主攻方向。
发明内容
本发明的目的是提供一种香青藤或其提取物在制备抗脑缺血药物中的应用，本发明首次发现香青藤及其提取物具有治疗脑缺血的成药潜力，可用于制备治疗脑缺血的药物，为脑缺血内科治疗开辟一条新途径，为脑缺血内科的治疗提供了新的药物来源。
为实现上述目的，本发明采用如下技术方案：
香青藤或其提取物在制备抗脑缺血药物中的应用。
所述香青藤(Illigera aromaticfa S.Z.Huang et S.L.Mo)为莲叶桐科青藤属香青藤的干燥藤茎，香青藤提取物为香青藤的水提物或香青藤的有机溶剂提取物。
所述香青藤水提物的提取方法为：
（1）取香青藤，加入5～7倍质量蒸馏水先浸泡45～75min，然后加热至80～100℃提取45～75min，过滤，收集滤液；
（2）滤渣再加入5～7倍质量蒸馏水加热至80～100℃提取45～75min，过滤，收集滤液；
（3）重复提取滤渣1～3次，合并多次提取得到的滤液并进行浓缩，即得。
所述香青藤的有机溶剂提取物为香青藤的乙醇提取物，提取方法为：
（1）取香青藤，加入5～7倍量体积浓度为85～95%的乙醇先浸泡45～75min，然后加热进行回流提取45～75min，过滤，收集滤液；
（2）滤渣再加入5～7倍量体积浓度为85～95%的乙醇，继续加热进行回流提取45～75min，过滤，收集滤液；
（3）重复提取滤渣1～3次，合并多次提取得到的滤液并进行浓缩回收乙醇，即得。
以上所述的香青藤提取物添加常规辅料，按照常规工艺制成临床可接受的用于预防或治疗脑缺血及其相关疾病的药物制剂。
所述制剂包括片剂、胶囊剂、滴丸或颗粒剂等。
所述的常规辅料包括淀粉、乳糖、微晶纤维素、糊精、磷酸钙、聚乙二醇-4000、聚乙二醇-6000、豆油、蜂胶、羧甲基纤维素钠、羟丙纤维素或交联聚维酮等中一种以上。
本发明的有益效果是：
1、本发明首次发现香青藤及其提取物具有治疗脑缺血的成药潜力，可用于制备治疗脑缺血的药物，为脑缺血的内科治疗开辟一条新途径，为脑缺血内科的治疗提供了新的药物来源。
2、本发明的香青藤资源丰富，提取制备方法简单易行，香青藤来源方便易得，可用于香青藤药材及其提取物的大量制备；成本低、污染小，利于节能减排条件下的大规模生产，产业化前景好。
3、本发明通过研究香青藤提取物对脑缺血再灌注损伤大鼠的治疗作用，证明了香青藤提取物可以改善缺血大鼠神经功能，降低大鼠脑梗死面积和脑水肿，有力证明了本发明的香青藤提取物对脑缺血损伤具有明显的拮抗作用，可用于制备抗脑缺血药物。
4、本发明的香青藤或香青藤提取物还具有祛风活血的功效，可用于风湿骨痛、跌打损伤、肥大性脊柱炎等症，可以在用于预防或治疗脑缺血的同时起到治疗其他合并症的效果。
5、香青藤可以与其他药材进行配伍组成复方药用于预防或治疗脑缺血，还可以将香青藤提取物与辅料混合制成片剂、胶囊、滴丸或颗粒剂，这些制剂较为方便服用，能单独或者搭配其他药剂服用，通过内服能将药性通过血液快速输送至病症部位进行有效治疗。
附图说明
图1香青藤提取物及其他组对脑缺血大鼠脑含水量的影响作用；
图2香青藤提取物及其他组对脑缺血大鼠神经行为学评分的影响作用；
图3香青藤提取物及其他组对脑缺血大鼠脑梗塞率的影响作用；
图4各组脑缺血大鼠脑TTC染色结果。
具体实施方式
为了更加详细的介绍本发明，下面结合实施例，对本发明做进一步说明。
实施例1 香青藤提取物的制备
取香青藤1kg，加入6L蒸馏水浸泡60min，然后加热至80℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至80℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至80℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.0g/mL，即得。
实施例2 香青藤提取物的制备
取香青藤1kg，加入6L蒸馏水浸泡60min，然后加热至90℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.1g/mL，即得。
实施例3 香青藤提取物的制备
取香青藤1kg，加入6L蒸馏水浸泡60min，然后加热至98℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至98℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至98℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.2g/mL，即得。
实施例4 香青藤提取物的制备
取香青藤1kg，加入7L蒸馏水浸泡75min，然后加热至80℃提取75min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；滤渣再加入5L蒸馏水加热至98℃提取45min，过滤，收集滤液；合并3次提取液，浓缩至1.0～1.2g/mL，即得。
实施例5 香青藤提取物的制备
取香青藤1kg，加入6L体积浓度为90%的乙醇先浸泡60min，然后加热进行回流提取60min，过滤，收集滤液；滤渣再加入6L体积浓度为90%的乙醇，继续加热进行回流提取60min，过滤，收集滤液；滤渣再加入6L体积浓度为90%的乙醇，继续加热进行回流提取60min，过滤，收集滤液；合并3次提取液并进行浓缩回收乙醇成膏，即得。
实施例6 香青藤提取物片剂的制备
【处方】由实施例3的方法制得的香青藤水提物（取相当于香青藤药材3kg的量）、干淀粉550g；
【制备方法】将所得的香青藤水提物、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，压制成1000片，所得的香青藤提取物片，每片含香青藤药材3g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上购买得到。
实施例7 香青藤提取物胶囊的制备
【处方】由实施例3的方法制得的香青藤水提物（取相当于香青藤药材3kg的量）、干淀粉550g；
【制备方法】将所得的香青藤水提物、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，分装成1000 粒，所得的香青藤提取物胶囊，每粒含香青藤药材3g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上售得到。
实施例8 香青藤提取物颗粒剂的制备
【处方】由实施例3的方法制得的香青藤水提物（取相当于香青藤药材10kg的量）、干淀粉550g；
【制备方法】将所得的香青藤水提物、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，分装成1000袋，所得的香青藤提取物颗粒，每袋含香青藤药材10g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上购买得到。
药性试验
为了验证本发明香青藤提取物的抗脑缺血损伤作用，本申请人进行了以下试验：
1、香青藤提取物对大鼠脑缺血再灌注损伤的治疗作用
供试品：所述香青藤提取物均是由实施例3中所述方法制备而成。
处理方法：
（1）大鼠大脑中动脉阻塞再灌注（MCAO/R）模型的建立
取SD大鼠按体重随机分4组：模型组、阳性药组（银杏二萜内酯葡胺注射液，1.25mg/kg）、香青藤组（1800mg/kg）、假手术组。假手术组和模型组两组给予等容积的溶剂，其余给药组每天灌胃给药1次，连续给药12天，于末次给药1h后开始造模。采用线栓法建立脑缺血/再灌注模型。线栓推进右颈内动脉，并堵塞脑中动脉，90min 后，将栓线移除，恢复血流，建立脑内再灌注状态。
（1）神经行为学评分
大鼠再灌后24 h进行行为学观察，按表1标准进行Bederson评分
（2）大鼠脑梗死面积和脑含水量的检测
戊巴比妥钠麻醉后取出全脑。将全脑置于-20℃冰箱冷冻20min。至大鼠脑变硬时，用锋利的刀片在视交叉及其前后各2mm处，做冠状切片，将脑组织切成五片，迅速将切好的脑片置于含有1% TTC的PBS缓冲溶液中，37℃避光温孵10min。10min后轻轻取出脑片，按顺序排列好，擦干脑片周围的水分，用数码相机进行拍照。数码照片使用Image J图像分析软件进行分析，计算梗死率的公式如下：
梗死率（%）=（缺血对侧脑面积-缺血侧非梗死区脑面积）/缺血对侧脑面积×100%
染色后的脑组织进行称重，此时重量为脑组织湿重，随后置于110℃烘箱24 h后称重，此时重量为脑组织干重，计算脑组织含水量的公式如下：
脑组织含水量（%）=（湿重-干重）/湿重×100%
（3）统计学方法
所有数据以平均值±标准差（mean ± SD）表示，spss软件（版本号19.0）进行统计分析。实验组间差异采用非参数检验对神经行为学评分进行统计分析，其余采用单因素方差分析（one-way ANOVA），当方差齐性时，选择LSD检验，当方差不齐性时，选择GamesHowell检验，P<0.05为差异有显著性，P<0.01为差异有极显著性。
结果：如图1-3所示，与假手术组相比，模型组大鼠神经行为学评分、脑梗死率和脑含水量极显著升高（P<0.01）。与模型组相比，阳性药组、香青藤组大鼠神经行为学评分显著降低（P<0.05，P<0.01）；阳性药组、香青藤组大鼠脑梗死率极显著降低（P<0.01，P<0.01）；阳性药组、香青藤组大鼠脑含水量极显著降低（P<0.01，P<0.01）。脑TTC 染色结果如图4 所示。
注：## P<0.01 vs.假手术组；** P<0.05，** P<0.01 vs.模型组。
Claims (4)
1.香青藤提取物在制备抗脑缺血药物中的应用，所述香青藤为莲叶桐科青藤属香青藤的干燥藤茎，香青藤提取物为香青藤的水提物；所述香青藤水提物的提取方法为：
（1）取香青藤，加入5～7倍质量蒸馏水先浸泡45～75min，然后加热至80～100℃提取45～75min，过滤，收集滤液；
（2）滤渣再加入5～7倍质量蒸馏水加热至80～100℃提取45～75min，过滤，收集滤液；
（3）重复提取滤渣1～3次，合并多次提取得到的滤液并进行浓缩，即得。
2.根据权利要求1所述的应用，其特征在于，香青藤提取物添加常规辅料，按照常规工艺制成临床可接受的用于预防或治疗脑缺血的药物制剂。
3.根据权利要求2所述的应用，其特征在于，所述制剂包括片剂、胶囊剂、滴丸或颗粒剂。
4.根据权利要求2所述的应用，其特征在于，所述的常规辅料包括淀粉、乳糖、微晶纤维素、糊精、磷酸钙、聚乙二醇-4000、聚乙二醇-6000、羧甲基纤维素钠、羟丙纤维素或交联聚维酮中一种以上。
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