CN113499377B - 一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物及其制备方法与应用 - Google Patents
一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物及其制备方法与应用 Download PDFInfo
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Abstract
本发明提供了一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物及其制备方法与应用，其包括：黄芪提取物、丹参提取物、玛俐那提取物、牛大力提取物、浒苔提取物、去核山楂提取物、桑椹提取物、去核大枣提取物、枸杞子提取物、乌梅肉提取物,上述中药配伍，共奏活血化瘀，益精养肝，化浊降脂之功效。药理实验表明，本申请的组合物能降低肝损伤大鼠的肝脏指数及血清中AST、ALT、LDH水平和高血脂大鼠血清中TC、TG和LDL‑C水平，升高HDL‑C水平，控制高脂模型大鼠体重增长，具有明确的保肝和辅助降血脂作用。
Description
技术领域
本发明涉及组合物及医药技术领域，具体涉及一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物及其制备方法与应用。
背景技术
近年来随着人们生活水平的提高，高血脂症的发病率逐年上升，高血脂症 (HLP)是指人体内脂肪代谢或运转异常导致血清总胆固醇(TC)、甘油三酯(TG) 和低密度脂蛋白胆固醇(LDL-C)水平升高，高密度脂蛋白胆固醇(HDL-C)水平降低。高血脂症容易引发冠状动脉粥样硬化、冠心病、心肌梗死、脑卒中、脂肪肝和胰腺炎等疾病，严重影响人们的生活质量和身体健康。
中医古文献并无“血脂”之名称,但在《黄帝内经》中已有“脂者”“油脂”“脂膜”等记载，《灵枢·逆顺肥瘦》说“广肩腋项，肉薄厚皮而色黑，唇临临然，其血黑以浊，其气涩以迟”，其中“血黑以浊”形象地描述了气血津液代谢失调，以致痰瘀胶结于血脉使血液浊滞的状态,与现代高脂血症、高粘血症的概念相近。根据临床表现,中医将高血脂归入“眩晕”“胸痹”“中风”“血瘀”“痰湿”等病症范畴。血脂形成、吸收、转运均和肝、脾关系密切。中药中的皂苷类、黄酮类、多糖类和蒽醌类等成分都具有辅助降血脂的功效，其作用机制可能与抗氧化作用、抑制外源性脂类和胆酸的吸收、抑制脂类物质的合成及减少脂类物质在血管内皮的沉积等有关。
化学性肝损伤是指由某些药物、环境中的化学有毒物质或者酒精等对肝脏造成的损害,会导致脂肪肝、肝纤维化、肝硬化和肝癌等疾病，以药物性和酒精性肝损伤最为常见。化学毒物损伤肝脏的机理主要有：抑制肝脏脂肪变性、抑制脂质过氧化反应和抑制胆汁淤积反应。肝脏对外来化学毒物生物转化和解毒功能主要包括以下四种方式：氧化解毒、还原解毒、水解解毒和结合毒性。肝脏在防御有害因素对机体损害的同时，自身也会受到伤害。
传统医学中对化学性肝损伤没有特定的病名记载,现代医家根据其“胁痛、右胁肋部肿块、黄疸、腹水”等临床表现,将其归为“胁痛、癥积、黄疸、鼓胀”等范畴。中医学认为化学性肝损伤的病因主要为“药毒、酒毒、化学及环境毒物”。中医药主要从消除自由基、降低谷丙转氨酶和谷草转氨酶、减少炎性因子的产生和改善肝脏病理组织等方面防治化学性肝损伤。中医药在治疗化学性肝损伤方面具有安全性高、适用性强、治疗效果显著、不加重肝脏负担、治疗疾病类型多样化等特点。
发明内容
本发明提供了一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物及其制备方法与应用。本发明所述的组合物的组方中，玛俐那、牛大力、浒苔、山楂、枸杞子、桑椹、乌梅和大枣为既是食品又是药品的果实类原料，黄芪和丹参为可用于组合物的中药，上述中药配伍，共奏活血化瘀，益精养肝，化浊降脂之功效。
本发明是通过如下技术方案实现的：
一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物，所述组合物包括：黄芪提取物9-30份、丹参提取物5-15份、玛俐那提取物6-15份、牛大力提取物6-15份、浒苔提取物6-15份、去核山楂提取物9-12份、桑椹提取物9-15份、去核大枣提取物6-15份、枸杞子提取物6-12份、乌梅肉提取物6-12份，辅料适量。
在本发明提供的一些实施方案中，一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物，由包括如下重量份数的成分制成：黄芪提取物10份、丹参提取物5份、玛俐那提取物10份、牛大力提取物10份、浒苔提取物10 份、去核山楂提取物9份、桑椹提取物9份、去核大枣提取物6份、枸杞子提取物6份、乌梅肉提取物6份，辅料适量。
在本发明提供的一些实施方案中，所述辅料包括魔芋胶、明胶、果胶、甜菊糖苷、木糖醇、山梨酸钾及水。
本发明还提供一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物的制备方法。
步骤(1)：浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药，加适量水浸泡后，置胶体磨中研磨处理后，提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，即得浒苔、去核山楂、乌梅、枸杞子、去核大枣、桑椹提取混合提取物；
步骤(2)：浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药，加适量的水，浸泡0.5～2h，煎煮10～30mi n，置胶体磨中研磨处理，即得浒苔、去核山楂、乌梅、枸杞子、去核大枣、桑椹提取混合提取物；
步骤(3)：黄芪提取物的制备：取适量黄芪，加适量水回流提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，即得黄芪提取物；
步骤(4)：丹参提取物的制备：取适量丹参，加适量水提取2-3次，每次 1-2h，滤过，合并提取液，减压浓缩，即得丹参提取物；
步骤(5)：玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用 90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得玛俐那提取物；
步骤(6)：牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用 90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得牛大力提取物；
将步骤(1)、步骤(3)-步骤(6)提取物混合均匀，加入适宜的辅料，制成所述的软糖；将步骤(2)-步骤(6)提取物混合均匀，加入适宜的辅料，制成所述的果糕、煎膏剂半固体或液体制剂。
在一个实施方案中，步骤(2)中加水量为各原料药总重量的9倍。
在一个实施方案中，步骤(1)、步骤(3)、步骤(4)提取工艺中加水量为各原料药总重量的12倍。
在一个实施方案中，步骤(2)中，浸泡2h，煎煮15min。
在一个实施方案中，步骤(1)中，提取温度为90℃，提取3次，每次1.5h。
在一个实施方案中，步骤(3)中，黄芪的提取温度为100℃，提取3次，每次1.5h。
在一个实施方案中，步骤(4)中，丹参的提取温度为80℃，提取2次，每次2h。
步骤(5)、步骤(6)提取工艺中被提取物与乙醇质量体积为各1g:5ml，每次浸提2h。
本发明还提供了所述组合物在制备用于辅助降血脂和对化学性肝损伤具有保护作用的保健品中的应用。
【原料药的功效和主治】
黄芪为豆科植物蒙古黄芪Astragalus membranaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao或膜荚黄芪Astragalus membranaceus (Fisch.)Bge.的干燥根。甘，微温。归肺、脾经。具有补气升阳，固表止汗，利水消肿，生津养血，行滞通痹，托毒排脓，敛疮生肌的功效。用于气虚乏力，食少便溏，中气下陷，久泻脱肛，便血崩漏，表虚自汗，气虚水肿，内热消渴，血虚萎黄，半身不遂，痹痛麻木，痈疽难溃，久溃不敛。
丹参为唇形科植物丹参Salvia miltiorrhiza Bge.的干燥根和根茎。苦，微寒。归心、肝经。具有活血祛瘀，通经止痛，清心除烦，凉血消痈的功效。用于胸痹心痛，脘腹胁痛，癥瘕积聚，热痹疼痛，心烦不眠，月经不调，痛经经闭，疮疡肿痛。
玛俐那为蔷薇科(Rosaceae)山植属(Crataegus)植物光叶山植(Crataegusdahurica Koenhne ex Schneid)的新鲜和/或干燥成熟果实，又名面果、涨肚黄、面都落等。其味酸甜可口，在民间常入药，有健脾消积、生津止渴之功效，多用于小儿积滞。该药材植物资源丰富，果实产量大，是重要的药食两用天然植物资源。玛俐那作为药食同源的植物资源，具有功能性食品特征，毒副作用小，其开发的产品可作为食品或保健品供适用人群日常食用，对病症的预防、治疗或辅助治疗均具有重要意义。
牛大力是豆科崖豆藤属美丽崖豆藤的干燥根，作为传统的药食两用中药材，在我国南方地区被广泛用于制作药膳、药酒、煲汤等。早期临床己证明其可治疗腰痛、肾虚、风湿性关节炎、腰肌劳损、慢性肝炎、病后体虚等多种慢性疾病。
浒苔((Enteromorpha prolifera,EP)，也称“海洋绿藻”或“绿藻”，为绿藻门石莼目石莼科浒苔属的藻类植物，其作为潮间带滩涂中的天然野生绿藻，自然繁殖能力强且产量巨大，具备很高的开发和利用价值。多糖为浒苔的主要活性成分，目前己有研究表明浒苔多糖具有降血脂功效。
山楂为蔷薇科植物山里红Crataegus pinnatifida Bge.var.MajorN.E. Br.或山楂Crataegus pinnatifida Bge.的干燥成熟果实。酸、甘，微温。归脾、胃、肝经。具有消食健胃，行气散瘀，化浊降脂的功效。用于肉食积滞，胃脘胀满，泻痢腹痛，瘀血经闭，产后瘀阻，心腹刺痛，胸痹心痛，疝气疼痛，高脂血症。
大枣为鼠李科植物枣Ziziphus jujuba Mill.的干燥成熟果实。甘，温。归脾、胃、心经。具有补中益气，养血安神。用于脾虚食少，乏力便溏，妇人脏躁。
枸杞子为茄科植物宁夏枸杞Lycium barbarum L.的干燥成熟果实。甘，平。归肝、肾经。具有滋补肝肾，益精明目的功效。用于虚劳精亏，腰膝酸痛，眩晕耳鸣，阳萎遗精，内热消渴，血虚萎黄，目昏不明。
桑椹为桑科植物桑Morus alba L.的干燥果穗。甘、酸，寒。归心、肝、肾经。具有滋阴补血，生津润燥的功效。用于肝肾阴虚，眩晕耳鸣，心悸失眠，须发早白，津伤口渴，内热消渴，肠燥便秘。
乌梅为蔷薇科植物梅Prunus mume(Sieb.)Sieb.etZucc.的干燥近成熟果实。酸、涩，平。归肝、脾、肺、大肠经。具有敛肺，涩肠，生津，安蛔的功效。用于肺虚久咳，久泻久痢，虚热消渴，蛔厥呕吐腹痛。
现代研究证实黄芪中主要含有多糖类、皂苷类、黄酮类等成分，可显著降低血浆总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)水平，增加胆固醇及胆汁酸排泄量，从而起到降血脂作用。其治疗肝损伤主要与蛋白质异二聚化活性、泛素蛋白连接酶结合、DNA结合转录激活活性、RNA聚合酶Ⅱ特异性、整合素结合等生物过程相关，还可以通过调节糖尿病并发症的晚期糖基化终末产物及其受体(AGE-RAGE)通路、卡波西(Kaposi)肉瘤相关疱疹病毒感染、乙型肝炎、肿瘤坏死因子通路等起到治疗肝损伤的作用。丹参中主要含有多糖类、丹参酮类和酚酸类等成分，通过阻止低密度脂蛋白氧化，降低脂质代谢酶活性和抑制胆固醇的合成起到降血脂的作用。其治疗肝损伤的机制可能与减少炎性因子生成，增强自由基清除酶的活性，抑制肝纤维化过程中ECM的生成，抑制肝星状细胞的活化和增殖，改善肝脏微循环等途径有关。玛俐那提取物包括甾醇类化合物、三萜类、脂肪醇苷类、芳香族化合物、单萜类化合物、苯丙素类等，玛俐那具有良好的体外自由基清除能力、铁离子还原能力和抗脂质过氧化能力，玛俐那具有良好的体内降血脂活性，具有开发成降脂药潜力。现代药理研究发现牛大力含有黄酮类化合物、生物碱类化合物等多种活性成分，改善脂代谢紊乱，具有降血脂、保肝等作用。浒苔中的多糖显著降低大鼠过氧化脂含量，具有一定抗衰老和抗血脂作用，其中硫酸多糖显著降低小鼠体内胆固醇与三酰甘油的含量，浒苔硫酸多糖可促进机体的免疫，加快了机体器官的物质代谢，肝内物质代谢降低了血脂在血液中的积累，起到降血脂的作用。山楂中主要含有有机酸、黄酮、三萜类等成分能显著降低血清胆固醇及甘油三酯水平，有效预防和治疗高血脂。枸杞子主要含有多糖类、糖脂类、苯丙素类和黄酮类等成分，通过降低血中总胆固(TC)、甘油三脂(TG)、低密度脂蛋白胆固醇(LDL-C)的水平以及降低肝内TC、TG水平起到治疗高血脂症的作用，其含有的甜菜碱可以起到对抗脂质过氧化，保护肝细胞膜的作用，治疗脂肪肝。桑椹中主要含有多糖、酚类、生物碱和氨基酸类等成分，具有降糖降脂、预防阿尔兹海默症、解酒、抑菌、提高免疫力、抗氧化、抗衰老等功效。大枣中含有糖类、蛋白质、生物碱、三萜、皂苷、黄酮、环腺苷酸、环鸟苷酸、氨基酸、维生素和矿物质等成分。大枣多糖能够降低血清总胆固醇、甘油三酯和动脉硬化指数(atherogenic index，AI)，同时抑制高密度脂蛋白胆固醇水平降低，起到降血脂的作用，通过增强机体单核细胞的吞噬功能、调节人体血清总蛋白与白蛋白水平、改善机体抵抗力和免疫力、清除自由基损伤、抗氧化和解毒作用治疗肝损伤。乌梅的化学成分主要为有机酸、萜类、甾醇、挥发性成分、氨基类、糖类、脂类、黄酮类和生物碱等，具有抗病毒性肝炎、抗肿瘤、抗菌消炎、抗疲劳、镇咳、镇静催眠及抗惊厥、抗变态反应、抗肿瘤、抗氧化、抗纤维化、降血脂、抑制黑色素、抗生育、治疗结肠炎、降血糖、防治结石和止血等药理作用。
本发明已经通过药理学实验证实了本发明的组合物能降低肝损伤大鼠的肝脏指数及血清中AST、ALT、LDH水平和高血脂大鼠血清中TC、TG和LDL-C 水平，升高HDL-C水平，控制高脂模型大鼠体重增长，具有明确的保肝和辅助降血脂作用。
与现有技术相比，本发明的优势是：配方科学，在归纳总结临床和实验研究文献的基础上，以现代医药学研究方法科学筛选优化了配方；用药安全性高，配方的原料以卫计委公布的既是食品又是药品类中药和可用于组合物黄芪和丹参组成；十种成分有机组合，全面兼顾，温性提取物有效中和了寒性提取物，有效控制了药物的烈性和毒性，降低了对消化系统的刺激，协同增效提高了辅助降血脂的效果。制备工艺先进，采用破壁机或胶体磨进行超微粉碎，药材利用率高，疗效好；辅料增效，如魔芋甘露聚糖、蜂蜜等对化学性肝损伤也具有保护作用，可以协同增效；剂型新颖，依从性好，尤其是软糖、果糕等剂型，极大改变了人们对组合物的感官认识，大大增加了消费者服用组合物的顺应性，让消费者在身心愉悦的同时起到了保健的作用。药理研究表明，其疗效确切、顺应可控，对高血脂症和化学性肝损伤均有改善作用，适合消费者长期食用，具有较大的开发价值。
附图说明
图1不同剂量对高血脂大鼠肝脏病理方面的影响(400×)；其中A：空白组， B：模型组，C：阳性组，D：低剂量组，E：中剂量组，F：高剂量组。
图2不同剂量对肝损伤大鼠肝脏病理方面的影响(400×)；其中：A：空白组， B：模型组，C：阳性组，D：低剂量组，E：中剂量组，F：高剂量组
具体实施方式
以下对本发明的优选实施例进行说明，应当理解，此处所描述的优选实施例仅用于说明和解释本发明，并不用于限定本发明。
(一)配方筛选试验例
1.实验材料
AK-400B中药粉碎机、KDM-1000调温电热套、AB265-S分析电子天平、 N1100-OSB-2100旋转蒸发仪、SHB-III循环水式多用真空泵、DZF-6050真空干燥箱、DLZY-0.5B真空冷冻干燥机、DNM-9602型多功能酶标仪、3-18R (3006004)型冷冻高速离心机、总胆固醇(TC)试剂盒、甘油三酯(TG)试剂盒、低密度脂蛋白胆固醇(LDL-C)试剂盒、高密度脂蛋白胆固醇(HDL-C) 试剂盒。
2.实验方法
2.1受试药物的制备
黄芪提取物的制备：取适量黄芪，加12倍量水回流提取3次，每次1.5h，滤过，合并提取液，减压浓缩后，冷冻干燥，即得黄芪提取物。
丹参提取物的制备：取适量丹参，加12倍量水提取2次，每次2h，滤过，合并提取液，减压浓缩，冷冻干燥，即得丹参提取物。
玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得玛俐那提取物；
牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得牛大力提取物；
浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药，加适量水浸泡后，置胶体磨中研磨处理后，提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，冷冻干燥，即得混合提取物。
2.2分组、给药及模型复制
取120只SD大鼠，随机分为空白对照组、模型对照组、阳性对照组，给药组1～9组，每组10只。其中，采用L9(34)正交试验设计给药组1～9组，以黄芪提取物、丹参提取物、八药食(浒苔+玛俐那+牛大力+山楂+大枣+乌梅+枸杞+桑椹)为影响因素，各设3个水平(临床用量的0.167、0.33、0.5倍)，参考现版《中国药典》中各药的用量，以60kg成人给黄芪提取物10g、丹参提取物5g、玛俐那提取物10g、牛大力提取物10g、浒苔提取物10g、去核山楂提取物9g、桑椹提取物9g、去核大枣提取物6g、枸杞子提取物6g、乌梅肉提取物6份g为临床用量，根据人和大鼠体表面积折算给药剂量。
表1因素水平表g/kg
2.3模型复制及指标检测
实验期间，实验动物自由饮水。空白对照组给予正常饲料，模型组、阳性组及低、中、高剂量组给予高脂饲料。连续灌胃给药30d，每天1次。空白对照组和模型对照组每天给予生理盐水。经腹腔注射7％水合氯醛溶液(4mL/kg) 麻醉后，腹主动脉取血，4℃离心机(3500r/min)离心15min，取上层澄清液体于-80℃保存，用于测定高胆固醇(TC)试剂盒、甘油三酯(TG)试剂盒、低密度脂蛋白胆固醇(LDL-C)试剂盒和高密度脂蛋白胆固醇(HDL-C)试剂盒；取新鲜的肝脏0.2g，加入预冷的生理盐水，制成10％的肝组织匀浆，3500rpm 离心10min，取上清液冷藏，用于测定谷草转氨酶(AST)试剂盒，谷丙转氨酶(ALT)试剂盒，总超氧化物歧化酶(SOD)试剂盒和丙二醛(MDA)试剂盒。
2.4数据分析
采用SPSS20.0对实验数据进行分析，实验数据采用平均值±标准差(x ±s)表示，组间比较采用方差分析；P＜0.05认为差异具有显著性，P＜0.01 认为差异具有极显著性。
2.5实验结果
各组血清的生化指标的结果见表2。
表2不同组方对高血脂大鼠血清中TC、TG、LDL-C和HDL-C水平的影响
注：与空白组比较，##P＜0.01；与模型组比较，**P＜0.01，P＜0.05
各指标进行综合评分作为正交试验评价指标。对各指标进行单一化处理，计算各给药组的综合降血脂作用。首先，对各指标进行标化，各指标的权重相等，当模型组的相应指标数值与正常组相比升高时(TC、TG、LDL-C)，V标化＝(V模型-V给药)/V模型×25/各给药组中最高分；当模型组的相应指标数值与正常组相比降低时(HDL-C)，V标化＝(V给药-V模型)/V模型×25/各给药组中最高分。V综合＝VTC+VTG+VLDL-C+VHDL-C。结果见表3、表4。
表3正交试验直观分析表
表4方差分析表
注：F0.05(2，2)＝19，F0.01(2，2)＝99
结果显示，与空白组相比，模型组血清中TC、TG、LDL-C、HDL-C水平均具有极显著性差异(P＜0.01)，说明造模成功。阳性对照组与模型对照组相比，具有极显著性差异(P＜0.01)。方差分析结果表明，黄芪、丹参、八药食对高血脂大鼠降脂作用均具有显著性影响(P＜0.05)，影响降脂药效的因素顺序为黄芪＞八药食＞丹参。直观分析表明，黄芪、八药食3水平最佳，丹参2水平最佳，因此确定最佳配伍组为A3B2C3，即黄芪、丹参、八药食用量分别为0.500g/kg、0.333g/kg、1.800g/kg。
(二)最优配方药效学实施例
实施例1：对高血脂大鼠降脂作用
1.实验材料：同上。受试药物为上述正交试验优化获得的具有最佳效果的中药提取物；正常对照组和模型对照组给予生理盐水；阳性对照组给予辛伐他汀。
2.给药剂量：阳性对照组(Y组)给予辛伐他汀5mg/kg；本申请组合物的低剂量组、中剂量组和高剂量组分别给予不同剂量的丹参-黄芪-八药食提取物 (0.250g/kg、0.165g/kg、0.900g/kg，0.500g/kg、0.333g/kg、1.800g/kg，1.000g/kg、0.667g/kg、3.600g/kg)；空白组与模型组给予生理盐水。
3.实验分组、给药、模型复制及指标测定：取60只雄性大鼠，随机分为6 组，空白对照组、模型对照组、阳性对照组、低剂量组、中剂量组和高剂量组，每组10只。实验动物饮食正常，自由饮水。实验期间，正常组给予正常饲料，模型组、阳性组及低、中、高剂量组给予高脂饲料，30d后，连续灌胃给药30d。经腹腔注射7％水合氯醛溶液(5mL/kg)麻醉后，腹主动脉取血，4℃离心机 (3500r/min)离心15min，取上层澄清液体于-80℃保存，用于测定高胆固醇 (TC)试剂盒、甘油三酯(TG)试剂盒、低密度脂蛋白胆固醇(LDL-C)试剂盒和高密度脂蛋白胆固醇(HDL-C)试剂盒。取以4％多聚甲醛固定好的新鲜肝脏，经过常规脱水、透明、浸蜡、包埋、切片后，进行HE染色，最后于400 倍光镜下观察肝脏病理形态的变化。
4.数据分析：采用SPSS20.0对实验数据进行分析，实验数据采用平均值±标准差(x±s)表示，组间比较采用方差分析；p＜0.05认为差异具有显著性，p＜0.01认为差异具有极显著性。
5.实验结果：见表5。
表5不同剂量对高血脂大鼠血清中TC、TG、LDL-C、HDL-C水平的影响
注：与空白组比较，##P＜0.01；与模型组比较，**P＜0.01
结果显示，与空白对照组比较，模型对照组大鼠血清中TC、TG、LDL-C、 HDL-C水平极显著升高(p＜0.01)；与模型对照组比较，各给药组均能极显著降低大鼠血清中TC、TG、LDL-C、HDL-C(p＜0.01)。
参见图1：结果显示，与空白组相比，模型组大鼠的肝细胞结构破坏，形状不规则，且大小不一，呈水肿及脂肪变性，细胞内可见明显弥漫脂肪空泡，炎性细胞浸润，散在灶状坏死；各给药组大鼠的肝细胞均有一定程度受损，但与模型组相比，肝细胞坏死及炎性细胞浸均有所改善，肝细胞结构较完整，排列更加有序，脂肪样改变面积减少。
实施例2：对肝脏的保护作用
1.实验材料：同上。受试药物为上述正交试验优化获得的具有最佳效果的中药提取物；正常对照组和模型对照组给予生理盐水；阳性对照组给予联苯双酯滴丸。
2.给药剂量：阳性对照组给予联苯双酯滴丸0.94mg/kg；本申请组合物的低剂量组、中剂量组和高剂量组分别给予不同剂量的丹参-黄芪-八药食提取物 (0.25g/kg、0.165g/kg、0.9g/kg，0.5g/kg、0.333g/kg、1.800g/kg，1.000g/kg、 0.667g/kg、3.600g/kg)；空白组与模型组给予蒸馏水。
3.实验分组、给药、模型复制及指标测定：取60只SD大鼠，随机分为6 组，空白对照组、模型对照组、阳性对照组、低剂量组、中剂量组和高剂量组，每组10只。连续灌胃给药30d，1次/d。末次灌胃给药7h后，空白对照组大鼠经尾静脉注射生理盐水(2mL/kg)，其余各组大鼠经尾静脉注射ConA (20mg/kg，以ConA质量计)。尾静脉注射ConA或生理盐水12h后，经腹腔注射7％水合氯醛溶液(4mL/kg)麻醉后，腹主动脉取血，4℃离心机(3500r/min) 离心15min，取上层澄清液体于-80℃保存，用于测定谷草转氨酶(AST)试剂盒，谷丙转氨酶(ALT)试剂盒和乳酸脱氢酶(LDH)试剂盒。
4.数据统计：采用SPSS20.0对实验数据进行分析，实验数据采用平均值±标准差(x±s)表示，组间比较采用方差分析；P＜0.05认为差异具有显著性，p＜0.01认为差异具有极显著性。
5.实验结果：见表6。
表6不同剂量对ConA致肝损伤大鼠血清中AST、ALT和LDH水平的影响
注：与空白组比较，##P＜0.01；与模型组比较，**P＜0.01
结果显示，与空白对照组比较，模型组大鼠血清中AST、ALT和LDH水平极显著升高(P＜0.01)；与模型对照组比较，各给药组大鼠血清中AST、ALT 及LDH水平均极显著降低(P＜0.01)。
参见图2，结果显示，与空白组比较，模型组大鼠的肝细胞出现灶状坏死、脂肪病变且肝门管区亦有大量炎性细胞浸润；与模型组比较，各给药组大鼠肝细胞的坏死情况均有改善。
(三)制备实施例
实施例1软糖的制备
一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物的原料药包括如下重量份数的成分：黄芪提取物10份、丹参提取物5份、玛俐那提取物10份、牛大力提取物10份、浒苔提取物10份、去核山楂提取物9份、桑椹提取物9份、去核大枣提取物6份、枸杞子提取物6份、乌梅肉提取物6份，辅料适量。
步骤(1)：浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药，加12倍水浸泡2h后，90℃回流提取3次，每次1.5h，滤过，合并提取液，减压浓缩，即得混合提取物。
步骤(2)：黄芪提取物的制备：取适量黄芪，加12倍水，100℃回流提取3 次，每次1.5h，滤过，合并提取液，减压浓缩，即得黄芪提取物。
步骤(3)：丹参提取物的制备：取适量丹参，加12倍水，80℃回流提取2 次，每次2h，滤过，合并提取液，减压浓缩，即得丹参提取物。
步骤(4)：玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用 90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得玛俐那提取物。
步骤(5)：牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得牛大力提取物。
步骤(6)：取1.0g果胶加40倍水浸泡2h，15g明胶加10倍水浸泡2h，加热熬煮至粘稠状态，将步骤(1)-步骤(5)制得的提取物加入到胶液中，加入1.0g 甜菊糖苷和0.1g山梨酸钾，于105℃加热熬煮，趁热浇模，即得。
步骤(7)：室温放置12h后，脱模。
步骤(8)：取脱模后的糕块包装，即得。
实施例2果糕的制备
一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物的原料药包括以下重量份的中药：黄芪提取物10份、丹参提取物5份、玛俐那提取物 10份、牛大力提取物10份、浒苔提取物10份、去核山楂提取物9份、桑椹提取物9份、去核大枣提取物6份、枸杞子提取物6份、乌梅肉提取物6份，辅料适量。
步骤(1)：浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药加各原料药总重量9倍量的水，浸泡2h，煎煮15min，置胶体磨中研磨，即得混合提取物。
步骤(2)：黄芪提取物的制备：取适量黄芪，加12倍水，100℃回流提取3 次，每次1.5h，滤过，合并提取液，减压浓缩，即得黄芪提取物。
步骤(3)：丹参提取物的制备：取适量丹参，加12倍水，80℃回流提取2 次，每次2h，滤过，合并提取液，减压浓缩，即得丹参提取物。
步骤(4)：玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用 90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得玛俐那提取物；
步骤(5)：牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用 90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得牛大力提取物；
步骤(6)：取10g魔芋胶加40倍水浸泡6h，加热熬煮至粘稠状态，将步骤 (1)-步骤(5)制得提取物加入到魔芋胶中，混匀，加入0.4g甜菊糖苷和0.1g 山梨酸钾，于120℃加热熬煮，趁热浇模，即得。
步骤(7)：室温放置2h后，脱模。
步骤(8)：取脱模后的糕块包装，即得。
实施例3煎膏剂的制备
一种具有辅助降血脂和对化学性肝损伤具有保护作用的组合物的原料药包括以下重量份的中药：黄芪提取物10份、丹参提取物5份、玛俐那提取物 10份、牛大力提取物10份、浒苔提取物10份、去核山楂提取物9份、桑椹提取物9份、去核大枣提取物6份、枸杞子提取物6份、乌梅肉提取物6份，辅料适量。
制备方法：
步骤(1)：浒苔、山楂、乌梅、枸杞子、大枣、桑椹提取物的制备：取适量上述中药加各原料药总重量9倍量的水，浸泡2h，煎煮15min，置胶体磨中研磨，即得混合提取物。
步骤(2)：黄芪提取物的制备：取适量黄芪，加12倍水，100℃回流提取3 次，每次1.5h，滤过，合并提取液，减压浓缩，即得黄芪提取物。
步骤(3)：丹参提取物的制备：取适量丹参，加适量水，80℃回流提取2 次，每次2h，滤过，合并提取液，减压浓缩，即得丹参提取物。
步骤(4)：玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用 90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得玛俐那提取物；
步骤(5)：牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用 90％乙醇水溶液浸提3次，被提取物与乙醇质量体积为各1g:5ml，每次浸提2h，滤过，合并提取液，减压浓缩，即得牛大力提取物；
步骤(6)：取步骤(1)-步骤(5)制得的提取物减压浓缩至清膏，加入清膏量1倍的中蜜，熬煮至挂旗、手捻出现拉丝和滴水成珠状态，即得。
步骤(7)：冷却至室温，包装即得。
显然，本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样，倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内，则本发明也意图包含这些改动和变型在内。
Claims (4)
1.一种具有辅助降血脂和对化学性肝损伤具有辅助保护作用的组合物，其特征在于，由如下重量份数的成分制成：黄芪提取物10-15份、丹参提取物5-10份、玛俐那提取物7-13份、牛大力提取物7-13份、浒苔提取物7-13份、去核山楂提取物9-10份、桑椹提取物9-13份、去核大枣提取物6-10份、枸杞子提取物6-10份、乌梅肉提取物6-10份，辅料适量；所述辅料包括魔芋胶、卡拉胶、黄原胶、明胶、果胶、麦芽糖醇、低聚半乳糖、甜菊糖苷、木糖醇、山梨糖醇、柠檬酸、山梨酸钾、蜂蜜中的一种或几种；所述组合物的剂型为软糖、果糕、煎膏剂；
所述组合物的制备方法，包括如下步骤：
步骤(1)：浒苔、去核山楂、乌梅、枸杞子、去核大枣、桑椹的混合提取物的制备：取适量上述中药，加适量水浸泡后，置胶体磨中研磨处理后，提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，即得浒苔、去核山楂、乌梅肉、枸杞子、去核大枣、桑椹的混合提取物；
步骤(2)：浒苔、去核山楂、乌梅、枸杞子、去核大枣、桑椹的混合提取物的制备：取适量上述中药，加适量的水，浸泡0.5～2h，煎煮10～30min，置胶体磨中研磨处理，即得浒苔、去核山楂、乌梅、枸杞子、去核大枣、桑椹的混合提取物；
步骤(3)：黄芪提取物的制备：取适量黄芪，加适量水回流提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，即得黄芪提取物；
步骤(4)：丹参提取物的制备：取适量丹参，加适量水回流提取2-3次，每次1-2h，滤过，合并提取液，减压浓缩，即得丹参提取物；
步骤(5)：玛俐那提取物的制备：将取适量干燥玛俐那，粉碎，25℃下用90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得玛俐那提取物；
步骤(6)：牛大力提取物的制备：将取适量牛大力，干燥、粉碎，60℃用90％乙醇水溶液浸提3次，滤过，合并提取液，减压浓缩，即得牛大力提取物；
将步骤(1)、步骤(3)-步骤(6)的提取物混合均匀，加入适宜的辅料，制成软糖；
将步骤(2)-步骤(6)的提取物混合均匀，加入适宜的辅料，制成果糕或煎膏剂。
2.如权利要求1所述的一种具有辅助降血脂和对化学性肝损伤具有辅助保护作用的组合物，其特征在于，由如下重量份数的成分制成：黄芪提取物10份、丹参提取物5份、玛俐那提取物10份、牛大力提取物10份、浒苔提取物10份、去核山楂提取物9份、桑椹提取物9份、去核大枣提取物6份、枸杞子提取物6份、乌梅肉提取物6份，辅料适量。
3.如权利要求1或2所述的一种具有辅助降血脂和对化学性肝损伤具有辅助保护作用的组合物，其特征在于，步骤(1)、步骤(3)、步骤(4)提取工艺中加水量为各原料药总重量的12倍，步骤(2)中加水量为各原料药总重量的9倍；步骤(1)中， 提取温度为90℃，提取3次，每次1.5h；步骤(2)中，浸泡2h，煎煮15min；步骤(3)中，黄芪的提取温度为100℃，提取3次，每次1.5h；步骤(4)中，丹参的提取温度为80℃，提取2次，每次2h。
4.如权利要求1或2所述的一种具有辅助降血脂和对化学性肝损伤具有辅助保护作用的组合物，其特征在于，步骤(5)、步骤(6)提取工艺中玛俐那或牛大力与乙醇质量体积为1g:5ml，每次浸提2h。
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