CN113750149B - 用于辅助抗氧化的中药复方制剂 - Google Patents
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Abstract
用于辅助抗氧化的中药复方制剂，由茯苓、陈皮、荷叶、山楂、甘草、白扁豆和黄芪组成，其中以重量份计，茯苓：10‑20份；陈皮：4‑14份；荷叶：2‑9份；山楂：3‑9份；甘草：1‑5份；白扁豆：1‑8份；以及黄芪：1‑8份。本发明的中药复方制剂可以作为抗氧化功能药食两用复方，尤其适用作绿色保健饮料产品例如护肝饮品。
Description
技术领域
本发明涉及一种可用于辅助抗氧化的中药复方制剂。
背景技术
人体在与外界环境不断的接触过程中，包括呼吸，污染，辐射等，均会产生大量自由基。尤其是人体受到外界不良因素损害时就会产生许多具有高度化学活性的自由基来抵御伤害，这些自由基的产生与清除处于一种动态平衡。但当机体处于疾病状态下时，这种平衡就会被打破，积累在体内的自由基不能被及时清除出去，就会引发机体产生炎症，进而可能引发多种疾病。
虽然目前很多慢性病的发生机制尚不甚明确，但很多学者认为与机体氧化应激反应有关，在预防慢性病时加强抗氧化治疗已得到普遍认可。此外，还有研究表明，体内自由基产生过多可能通过一系列生理生化反应引发心血管相关疾病和2型糖尿病。由此可见，各类氧化应激反应与慢性病的发生发展具有高度相关性。
因此，抗氧化逐渐成为预防和治疗各类慢性疾病的重要手段。抗氧化物质的作用则是能够直接作用于自由基或消耗掉易产生自由基的物质，以减少大量自由基对机体的伤害。由于实践证明，药用植物活性成分具有显著的抗氧化作用，因此合适的中草药复方在抗氧化活性或调节机体内氧化应激反应上无疑具有深远应用前景。
发明内容
本发明的目的是提供一种可药食两用的抗氧化中药复方制剂。
根据本发明的第一方面，提供了一种用于抗氧化的中药复方制剂，由茯苓、陈皮、荷叶、山楂、甘草、白扁豆和黄芪组成，其中以重量份计，茯苓：10-20份；陈皮：4-14份；荷叶：2-9份；山楂：3-9份；甘草：1-5份；白扁豆：1-8份；以及黄芪：1-8份。
本发明的复方制剂可以采用多种剂型，例如水提原液形式或添加辅料并干燥后制成的颗粒冲剂或胶囊等形式。辅料可以采用可溶性淀粉和糊精混合物等。
根据本发明的另一方面，提供了一种用于辅助抗氧化的保健饮品，例如护肝饮品，其包含上述中药复方制剂。
发明人通过多年精心筛选，形成了一种安全有效的中药降脂复方，其中茯苓为君，燥湿健脾；陈皮、黄芪为臣，补气固表、行气祛湿；荷叶、山楂、白扁豆为佐，健脾行气、化湿和中；甘草为使，健脾、调和诸药，七味药相辅相成，合而为一，扶正祛邪，利用身体自身之正气，通过实效运化，祛湿排毒，从而达到抵抗外界氧化应激反应的能力。
附图说明
图1为本发明的抗氧化评价实验流程示意图；
图2A为实验周期内各组小鼠体质量的变化趋势曲线图；图2B为实验结束时各组小鼠体质量柱形示意图；图2C为实验结束时各组小鼠的肝脏指数柱形示意图；
图3A和3B分别为各组小鼠肝脏脂质的不同过氧化产物水平柱形示意图；
图4A和4B分别为各组小鼠肝脏的不同抗氧化酶活性柱形示意图；
图5为各组小鼠肝脏抗氧化物质含量柱形示意图；以及
图6为各组小鼠肝脏组织病理学切片电子显微镜图。
所有图中统计学显著性分析表示为：a，与正常空白组比较，p＜0.05；b，与羧甲基茯苓多糖组比较，p＜0.05。
具体实施方式
下面结合具体实例和附图对本发明做进一步说明，本领域技术人员应该理解，实例和附图只是为了更好地理解本发明，并不用来做出任何限制。
制备中药复方原液
按重量份称取茯苓150g、陈皮90g、荷叶50g、山楂50g、甘草30g、白扁豆50g以及黄芪50g(均为市购)，加入纯净水4700mL，常温浸泡1h，然后常压下加热煮沸，计时0.5h，将提取液室温放凉后，采用8层纱布进行过滤，收集滤液备用，滤渣再加入4700mL饮用水，重复提取1次，即同一批原料先后共提取2次，第二次提取后，弃掉滤渣，合并2次的提取液，减压浓缩后获得4700mL原液。
制备复方颗粒制剂
向上述原液中加入329g混合辅料：可溶性淀粉和糊精混合物(二者质量比为4:1)，搅拌均匀后喷雾干燥，制得可例如用于冲服的颗粒制剂或固体饮料粉末248g。
复方颗粒制剂营养性能测试
将上述制备的复方颗粒制剂进行溶解度、粒度、分散性、冲调稳定性、堆密度、休止角、水分和吸湿性等各项指标检测，结果如表1所示。从表中可以看到抗氧化复方制剂各项指标要求为：溶解度约为330s；粒度要求均能通过六号筛；分散性要求均能通过筛网；冲调稳定性要求离心后无沉淀；堆密度值约为0.34；休止角约为29.4；水分约为2.08％和吸湿性约为15.87％。
表1表1保肝复方制剂理化指标测定结果
作为保健食品服用，上述制备的固体饮料粉末的成人推荐剂量可以为：每日二次，每次20g。
乙醇氧化损伤动物建模对照实验
实验材料
小鼠8-异构前列腺素(8-isoprostane)酶联免疫分析(ELISA)，货号ml058199-1，购自上海酶联生物科技有限公司；丙二醛测试盒，货号A003-1；总超氧化物歧化酶测试盒，货号A001-1；谷胱甘肽过氧化物酶测试盒，货号A005；微量还原型谷胱甘肽试剂盒，货号A006-2，均购自南京建成生物工程研究所。
受试物制备
中药复方原液的制备：各药材以按比例总共称重203.04g(茯苓64.8g,陈皮38.88g,荷叶21.6g,山楂21.6g，甘草12.96g，白扁豆21.6g，黄芪21.6g)，按照上述复方原液制备方法，得到复方原液216mL，冷藏于4℃冰箱，备用。
复配多糖(500mg/kg·BW)溶液配制：准确称取茯苓粉末，灵芝多糖，黄芪多糖，甘草多糖，党参多糖粉末100，200，300，200，100mg，混匀，溶于36mL蒸馏水中，为10天灌胃量，备用。
实验动物与分组
ICR小鼠：雄性，25-28g，48只；由中国医学科学院药用植物研究所动物实验中心提供，许可证编号：SYXK(京)2017-0020。
SPF级小鼠适应性喂养1周后，根据体质量水平，随机分组。各组均给予维持型饲料，自由饮食饮水。饲养温度为20～25℃，相对湿度55％～70％，昼夜明暗交替时间为12h/12h。
实验周期内30天，各组除灌胃受试物不同，其余处理均一致。末次灌胃后，除正常空白组外，模型空白组和复配多糖组小鼠禁食16h，然后1次性经口灌胃，给予50％乙醇12mL/kg BW，6h后取血取肝脏。
具体处理流程步骤见图1。
实验方法
将小鼠按照体质量水平，随机分为正常空白组(CK)，模型空白组(MD)，复配多糖组(500mg/kg·BW)，中药复方组(9.4g/kg·BW，生药量；C1)，共计4组，每组12只。其中，正常和模型空白组均灌胃同体积蒸馏水；复配多糖组灌胃多糖的复配溶液；中药复方组灌胃上述复方原液。
空白组和模型组，经口灌胃等体积蒸馏水，复配多糖组与中药复方组分别灌胃等体积受试物样品溶液，灌胃容量为0.2mL/10g，连续30d，每周记录小鼠体质量，并根据体质量来调整灌胃剂量，灌胃期间自由取食和饮水。
末次灌胃后，除空白组外，其他组禁食不禁水16h，在禁食10h后，一次性大剂量灌胃给予50％浓度的乙醇溶液，灌胃剂量为12mL/kg·BW，6h后水合氯醛注射麻醉后，进行取摘眼球血。麻醉摘眼球取血，1.5mL EP管收集血液，3000rpm/min离心15min，取上清液，-20℃冷冻备用。脱颈处死小鼠后，进行解剖，取肝脏，预冷生理盐水清洗去除残血，滤纸擦干，称重记录，然后将肝脏分离成2份，其1放入10％福尔马林固定液中，用于后续进行组织病理学观察，其2先放入冻存管中，后放入-80℃冰箱中储存备用，用于检测抗氧化功能指标；后收集小鼠肠道内存留的新鲜粪便，放入冻存管中，先置于液氮中预冷冻，后放入-80℃冰箱中储存备用，用于检测复配多糖对小鼠肠道微生物菌群的影响。
⑴抗氧化功能指标(MDA、8-Isoprostane、T-SOD、GSH-PX、GSH)检测：分别对实验小鼠肝脏组织和血清，进行操作检测，记录数据；
⑵进行组织HE染色切片制作和观察。
实验结果分析
(1)经过30天灌胃的实验周期，每周对小鼠体质量进行监测，绘制实验周期内各组小鼠体质量的变化趋势，结果如图2A所示。由于乙醇氧化损伤模型的造模时间是在实验结束前6h进行，所以在小鼠体质量的监测过程中，正常空白组和模型组小鼠处理一致，因此，在体质量变化趋势上，各个组较为一致，实验结束前的体重分析也没有显著性差异(P＞0.05，图2B)。经大剂量乙醇灌胃造模后，取小鼠肝脏进行称重并计算肝脏指数，结果如图2C所示，摄入大量乙醇的组别小鼠肝脏显著减小，导致其肝脏指数与正常空白组相比，显著降低(P＜0.05)。
(2)8-异前列腺素和丙二醛MDA均代表了体内脂质过氧化产物的累积，其含量越多，表明机体抗氧化能力越弱。如图3A和3B所示，经乙醇灌胃造模后，与正常空白组比较，模型空白组小鼠肝脏内的8-异前列腺素和MDA含量均显著升高(P＜0.05)，说明造模成功。与模型组比较，中药复方组小鼠肝脏内的8-异前列腺素含量显著下降(P＜0.05)，其中复配多糖组恢复到与正常空白组差异不显著的程度，说明在降低MDA含量角度，中药复方组下降差异显著(P＜0.05)，效果优于复配多糖。
(3)超氧化物歧化酶SOD和谷胱甘肽过氧化物酶GSH-Px是机体内重要的抗氧化酶类，具有清除体内氧化应激反应产生的过多的自由基作用，其含量越高抗氧化能力越强。如图4A和4B所示，与正常空白组比较，摄入大量乙醇的模型组及各受试物组小鼠肝脏内2种抗氧化酶的活性均显著降低(P＜0.05)，说明造模成功。与模型空白组相比，中药复方组小鼠肝脏内的2种酶活性均有显著提升(P＜0.05)，虽然与正常空白组还有一定差距，但表明本中药复方对改善因乙醇造成的机体氧化应激反应紊乱有明显正向作用。
(4)还原型谷胱甘肽GSH是机体内存在的还原性物质，具有清除氧自由基的作用，其含量越高说明机体抗氧化能力越强。如图5所示，大量乙醇摄入造模后，模型组较正常组小鼠肝脏中的GSH含量显著下降(P＜0.05)，亦说明造模成功。与模型空白组比较，灌胃复配多糖和中药复方组的小鼠肝脏内GSH含量明显上升(P＜0.05)，表明中药复方的摄入有助于缓解乙醇造成的氧化损伤，且以正常空白组差异不显著(P＞0.05)，说明本中药复方有助于模型小鼠机体的抗氧化能力恢复至正常水平。
(5)实验周期结束时，取小鼠肝脏，一部分用于进行生化指标检测，一部分用于制作HE病理切片，以观察小鼠肝脏的生理状态，进一步印证生化指标的检测结果。如图6所示(其中，CK：空白组；MD：模型组；FPDT：复配多糖组；C1：中药复方组)，各组小鼠的肝脏组织切片中均有炎症因子出现，其中，正常组小鼠肝脏中的炎症因子数目较少，肝细胞数目多且形态饱满；乙醇氧化损伤模型组小鼠肝脏中的炎症因子分布密集，组织中出现空泡，肝细胞数目减少且大小不一；灌胃中药复方的小鼠炎症因子有所减少，组织内空泡现象得到缓解，肝细胞数目增多且大小均匀，结果表明，乙醇氧化损伤模型造模成功，且本中药复方有助于缓解模型造成的肝脏损伤。
由此可知，本发明的中药复方制剂可以作为(辅助)抗氧化功能药食两用复方，尤其适用作绿色保健饮料产品例如护肝饮料。
Claims (4)
1.一种用于抗氧化的中药复方制剂，由茯苓、陈皮、荷叶、山楂、甘草、白扁豆和黄芪组成，其中以重量份计，茯苓：15份；陈皮：9份；荷叶：5份；山楂：5份；甘草：3份；白扁豆：5份；以及黄芪：5份。
2.根据权利要求1的复方制剂，其中该制剂为水提液形式。
3.根据权利要求1的复方制剂，其中该制剂为颗粒冲剂或胶囊形式。
4.一种用于辅助抗氧化的保健饮品，包含权利要求1的中药复方制剂。
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