CN115487280B - 一种用于解酒保肝、促进肝再生的中药组合物及其制剂和制备方法 - Google Patents
一种用于解酒保肝、促进肝再生的中药组合物及其制剂和制备方法 Download PDFInfo
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Abstract
本发明公开了一种用于解酒保肝、促进肝再生的中药组合物及其制剂和制备方法，属于中药制药技术领域。一种用于解酒保肝、促进肝再生的中药组合物，所述中药组合物由以下重量份数配比的组分组成：葛根提取物4‑6份、炒白芍提取物2‑3份、熟地黄提取物4‑5份、丹参提取物2‑4份、姜黄提取物1‑3份、五味子提取物1‑3份、玄参提取物2‑4份、炙甘草提取物1‑3份、炒黄芩提取物2‑3份、枳椇子提取物2‑4份、藤茶提取物2‑3份、桑黄提取物2‑4份。本发明提供的中药组合物安全性和药效实验发挥抗ALD和促进肝再生的疗效。
Description
技术领域
本发明涉及中药制药技术领域，更具体地说，涉及一种用于解酒保肝、促进肝再生的中药组合物及其制剂和制备方法。
背景技术
酒精性肝病（Alcoholic liver disease, ALD）包括脂肪肝、酒精性肝炎、肝纤维化和肝硬化，严重者可发展为肝癌；ALD发病率逐年增长，已成为当今世界范围内一个重要的公共卫生问题。流行病学调查显示酒精是全球范围内导致肝癌发生、死亡的次要因素。迄今为止，ALD的临床治疗仅停留于戒酒、营养补充、改善症状层面，且干预靶点单一，疗效并不确切。近年来研究发现，包括ALD在内许多慢性肝病患者存在肝脏再生功能障碍现象，若伴有肝脏结构异常和明显的肝纤维化时，患者往往预后不良。现代医学认为ALD疾病进展过程中伴随着肝再生障碍，临床研究发现急性酒精或者慢性酒精摄入可以抑制部分肝切除术后诱导的肝细胞增殖反应。而且，酒精性肝炎住院病人中，肝细胞增值能力的下降与其病死率上升之间有相关性。由此可见，促进肝再生是延缓各类型慢性肝病的疾病进展和改善预后的关键。
中医治疗ALD具有丰富的临床经验，中医理论认为ALD的肝再生障碍属于“虚证”范畴，其发生是由于饮酒过度，导致湿热内蕴，影响及脾，致脾运化失健，痰湿内生，肝疏泄不调，从而气机郁结，血行不畅而瘀滞于内，最终气滞、痰湿、淤血互结，积于胁下而发病；其发病机理与痰、湿、淤等密切相关，主要责之于肝、脾两脏；治则以祛湿化浊、疏肝健脾、清热化湿、活血化瘀和祛痰散结为主。中医中肝肾同源的理念也为肝再生提供了新思路；《内经》提出“髓生肝”理论，中医“生髓”治疗多种病证的疗效机制之一是通过影响干细胞及其组织微环境。“生髓”即“使髓生”，维护或促进“髓”处于正常的生理状态。“虚”、“损”、“积”是“髓失生肝”的重要病机转换，三者之间通过肝再生修复机制存在如下关系: 肝损伤后通过正常肝再生修复而不会形成虚证，肝再生修复机制受到干扰后会出现虚证与实证两种结果，并会形成“因虚致实”、“因实致虚”恶性病理循环。肝再生紊乱是导致肝纤维化向肝硬化进展中出现“因实致虚”( “积生虚损”) 、“因虚致实”( “虚损生积”) 两种病机转换( “虚积互生”) 的关键环节。治疗主张协调运用“扶正除积”与“除积复正”，“补肾生髓成肝”是其重要治疗法则之一。
在急慢性肝病病程进展中，肝再生是重要而关建的病理生理学基础，维持正常的肝再生是修复肝损伤的必然机制，肝再生失调与肝衰竭、肝硬化和肝癌的发生发展密不可分。中医药广泛用于急慢性肝病的治疗，其作用机制可能是多途径、多层次、多***、多靶点、多时限地调控肝再生过程。近年来研究发现，肝再生障碍可能与慢性肝损伤中肝细胞呈现衰老状态，自我再生能力受损密切相关。中医药是我国得天独厚的医药卫生资源，在多种慢性疾病治疗中彰显出独特的优势，已成为解酒保肝和促进肝再生药物开发的热点领域。
发明内容
1．要解决的技术问题
针对现有技术中存在的问题，本发明的第一个目的在于提供一种用于解酒保肝、促进肝再生的中药组合物，本发明的第二个目的在于提供中药组合物制剂，本发明的第三个目的在于提供中药组合物的制备方法；本发明提供的中药组合物安全性和药效实验发挥抗ALD和促进肝再生的疗效。
2．技术方案
为解决上述问题，本发明采用如下的技术方案。
为了实现上述第一个目的，本发明采用了以下的技术方案：
一种用于解酒保肝、促进肝再生的中药组合物，所述中药组合物由以下重量份数配比的组分组成：葛根提取物4-6份、炒白芍提取物2-3份、熟地黄提取物4-5份、丹参提取物2-4份、姜黄提取物1-3份、五味子提取物1-3份、玄参提取物2-4份、炙甘草提取物1-3份、炒黄芩提取物2-3份、枳椇子提取物2-4份、藤茶提取物2-3份、桑黄提取物2-4份。
进一步的，将葛根提取物4份，炒白芍提取物2份，熟地黄提取物5份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份。
为了实现上述的第二个目的，本发明采用了以下的技术方案：
一种用于解酒保肝、促进肝再生的中药组合物制剂，所述中药制剂包括权利要求1所述的中药组合物和/或适量的其他原料。
进一步的，所述中药制剂为片剂、咀嚼片、粉剂、散剂、颗粒剂、冲剂、胶囊剂、膏剂、丸剂或液体制剂。
进一步的，所述中药制剂还包括食品添加剂。
为了实现上述的第三个目的，本发明采用了以下的技术方案：
一种制备中药组合物的方法，所述方法包括如下步骤，先将所述中药组合物中的葛根、炒白芍、熟地黄、丹参，姜黄，五味子，玄参，炙甘草，炒黄芩，枳椇子，藤茶及桑黄的提取物通过以下方法获得：
1）将葛根实体破碎，加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到葛根提取物粉末；
2）将炒白芍破碎，加入3-7倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炒白芍提取物粉末；
3）将熟地黄加入4-6倍量水煎煮4-6次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到熟地黄提取物粉末；
4）将丹参加入3-7倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；浸膏冷冻真空干燥，粉碎过六号筛，得到丹参提取物粉末；
5）将姜黄加入1-4倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到姜黄提取物粉末；
6）将五味子加入1-4倍量水煎煮1-2次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到五味子提取物粉末；
7）将玄参加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到玄参提取物粉末；
8）将炙甘草加入1-4倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炙甘草提取物粉末；
9）将炒黄芩加入3-8倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炒黄芩提取物粉末；
10）将枳椇子加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到枳椇子提取物粉末；
11）将藤茶加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到藤茶提取物粉末；
12）将桑黄加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到桑黄提取物粉末；
13）最后将葛根，炒白芍，熟地黄，丹参，姜黄，五味子，玄参，炙甘草，炒黄芩，枳椇子，藤茶，桑黄的这12种提取物的浸膏干粉按照一定比例混合。
然后，将葛根，炒白芍，熟地黄，丹参，姜黄，五味子，玄参，炙甘草，炒黄芩，枳椇子，藤茶，桑黄的这12种提取物的浸膏干粉按照一定比例混合。
葛根为豆科植物野葛 Puearia lobata（Willd.）Ohwi.的干燥根，具有发表解肌、升阳透疹、解热生津的功效，入脾、胃二经。葛根是我国最常见的中药之一，具有悠久的临床使用历史。其始载于我国汉代的《神农本草经》，梁代陶弘景《本草经集注》曾云：“即今之葛根，人皆蒸食之”，如今葛根被列入中华人民共和国国家卫生健康委员会发布的《按照传统既是食品又是中药材的物质目录》。在历代文献记载中葛根对“酒病”的治疗有广泛的应用，如《普济方》第253卷所载的“解酒毒方”中，采用葛根者有15首。近几十年来，诸多学者已从葛根中分离鉴定出 100 余个化合物，包括黄酮、三萜、香豆素、有机酸等类成分。葛根具有广泛的药理作用，如保肝、抗肿瘤、神经保护、心脏保护、改善胰岛素抵抗、抗炎、抗氧化等。现代研究表明，葛根能够显著降低慢性酒精诱导的小鼠血清中ALT和AST和肝组织γ-GT水平升高，提高肝脏GSH含量，同时增强肝脏抗氧化能力。
白芍（paeonia lactiflora）味苦酸甘，性微寒，药用根，归肝、脾经。《本经》记载，白芍“主邪气腹痛、除血痹、破坚积寒热、利小便、益气”。白芍总苷为白芍中的重要药理成分，可通过减少肝组织中NF-KB的含量，抑制炎症反应，保护肝细胞。炒白芍是白芍用文火加热，炒至表面微黄色，取出放凉的方法制成的，具有养血柔肝，缓中止痛，敛阴收汗，治胸腹胁肋疼痛的作用。临床用炒白芍民间用于治疗风湿冷痛、风湿关节炎、腰肌劳损、体虚神衰、四肢抽搐、慢惊风、犬咬伤等病症。相对于白芍，炒白芍药性更为温和，能平抑肝热，因而在肝再生方面，选用炒白芍，更有利于肝脏修复。
地黄是玄参科、地黄属多年生草本植物。以新鲜或干燥块根入药。熟地黄为其经酒蒸煮炮制后所得的一味中药，《中华本草》称其为补益药。熟地黄微温，味甘，归肝、肾经，具有滋阴补血，益精填髓的功效。常用于肝肾阴虚、内热消渴。在培精滋阴方面，六味地黄丸、左归丸、金匮肾气丸等常用方剂皆以熟地黄为君。现代医学证实，熟地黄中地黄素A、地黄素D等成分表现出较明显的保肝活性，具有显著促进造血、抗氧化应激、增强免疫力的功能。
丹参为唇形科植物（Salvia Miltiorrhiza Bge）的干燥根及根茎具有祛瘀止痛、活血通经、清心除烦之功，味苦，微寒。近年来通过对丹参制剂基本药理作用的实验研究和临床验证，发现其对肝病的营养调理作用是基于改善肝脏及全身血液循环，促进肝细胞再生，改善纤维化，恢复肝功能，解毒、调节免疫功能和抗肿瘤等作用。其中丹参素为其有效成分之一，具有扩张血管改善微循环选择性作用于受体操作性钙通道，防止细胞钙超载的作用。丹参素能有效降低体内过氧化物歧化酶（SOD）活性，能够抑制脂质过氧化，保护线粒体功能，增加肝细胞蛋白质合成功能，进而促进肝细胞再生。另外丹参素可降低ALT含量，使血浆纤维联结蛋白（PFN）升高，提高其网状内皮***的吞噬功能及调理素活性，也可增强巨噬细胞的免疫活性，从而防止肝脏的免疫损伤。有研究发现，丹参能增加损伤肝细胞 DNA、RNA及蛋白质的含量并促进 DNA 的合成，促进铜蓝蛋白的合成，有利于促进肝细胞再生与修复。其中丹参对肝脏 DNA 合成具有2个特点：（1）作用在 DNA合成的持续阶段；（2）不改变DNA 合成的规律性。丹参的主要化学成分有30多种其脂溶性成分大多为醌型红黄色物质如丹参酮Ⅰ、ⅡA、ⅡB，丹参醌 A、B、C，丹参酸钾脂，异丹参酮Ⅰ、Ⅱ，隐丹参酮，异隐丹参酮等；其水溶性成分大多为酚性醛、丹酚酸A、丹酚酸B、二萜酸如丁二酸丹参酸甲、乙、丙及3，4-二羟基苯甲酸等此外还分离出β-谷甾醇、维生素E等。而丹参素的提取在药物制取的过程中极为重要，通过水煎煮法，索氏提取法和超声提取法的实验对比得出，用索氏提取法提取出的丹参素纯度最高。
姜黄是姜科植物姜黄 Curcuma longa L的根茎。姜黄味苦、辛，性温，归肝、脾经，具有行气破淤、通经止痛的功效。姜黄素是从中药姜黄中提取的一种酚类色素，具有抗炎、抗氧化、抗缺血和抗肝纤维化等广泛的药理作用。在促进肝再生方面，姜黄素能够抑制TNF-α的过度表达以抑制炎症反应。在保肝机制方面，姜黄素可以从多个途径来抗纤维化：1.通过ERK 途径部分降低缺氧诱导因子-1α的表达，预防肝纤维化，2.通过抑制 MCP-1 来减少Gr1hi 单核细胞的募集，从而保护肝脏免受诱导致纤维化。
五味子为木兰科植物五味子Schisandra chinensis（Turcz.）Baill（***，以达到抗炎目的；五味子甲素（Sin A）能显著增加表皮生长因子受体（EGFR）、细胞周期蛋白D1、细胞周期蛋白D依赖激酶4（CDK4）和PCNA蛋白的表达，抑制细胞周期抑制剂p21和p27的表达，进而显著促进肝脏再生。
玄参为玄参科玄参的干燥根，始载于《神农本草经》，长于滋肾养阴，填精益髓，解毒散结，为甘、苦、咸、性微寒，具有滋阴、降火、除烦、解毒的功能。近年来的研究发现不同种玄参提取物包括：中苯丙素，乙酸乙酯萃取物等都具有肝脏保护活性。刘冠璋等学者做出研究表明玄参水提物能够增强SOD和GSH-Px的活性，同时对血清中TBiL活性具有一定的降低作用，对CCL4所致大鼠急性肝损伤具有明显的保护作用。
甘草，是一种中医药学当中的常见补气类药用植物。其性味甘平，具有清热解毒，补中益气，缓急止痛，调和药性等作用。甘草中大量的黄酮，多糖具有抗氧化效应，能够有效的对氧自由基进行清除，从而保护细胞膜免受损伤；除此之外，甘草中的甘草酸和甘草次酸能够通过抑制脂加氧酶和磷脂酶A2的活性，达到降低PGs合成释放的目的，起到抗炎的效果，因而甘草被广泛用于各类肝炎的临床治疗中。
黄芩，别名山茶根、土金茶根，是唇形科植物黄芩Scutellaria baicalensisGeorgi的干燥根，主要有黄芩苷、汉黄芩苷、黄芩素、汉黄芩素等成分，始记载于《神农本草经》，具有“诸热黄胆，肠泄痢，逐水，下血闭，恶疮疽蚀火疡”的功效，传统用于治疗中上焦湿热、疮疡。现代研究表明，黄芩具有抗肝炎、抗肿瘤，抗氧化、护肝等多项活性作用。研究表明有一种途径汉黄岑素通过诱导中性粒细胞凋亡，减少肝脏中性粒细胞浸润，从而减轻免疫性肝损伤。另一种途径是黄芩苷可能通过抑制内质网应激凋亡通路而减轻乙醇诱导的大鼠肝细胞过度凋亡。此外，黄芩叶作为黄芩的非药用部位，同样可以通过增强ADH活性来改善酒精代谢，从而缓解酒精诱导的小鼠醉酒症状，同时通过逆转氧化应激来保护酒精对小鼠肝脏的急性损伤。
枳椇子为鼠李科枳椇属植物的成熟种子。本品属养阴，生津，润燥，止渴，凉血类药物。具有解酒毒，止渴除烦，止呕，利小大便之功效。枳椇子解酒的奇效早在《食疗本草》中便已阐明“昔有南人修舍此木，误落一片入酒瓮中，酒化为水也”。现今的研究发现枳椇子中大量的黄酮、多糖等抗氧化物质，不仅增强乙醇脱氢酶活性，增强乙醇代谢(参见:于刚,***,曹晓钢.枳椇子活性多糖的提取工艺及解酒功能研究[J],广西轻工业, 2 0 0 7 , 10:3 ~ 4.）；还可以提高抗氧化物GSH水平，抗脂质过氧化从而减轻肝脏内炎性细胞的浸润程度；不仅如此，其中的二氢杨梅素经过研究发现对于肝细胞再生也有重要功效。
藤茶，别名霉茶，是葡萄科蛇葡萄属植物显齿蛇葡萄Ampelopsis grossedentata(Hand-Mazz) W.T.Wang的嫩茎叶，作为中药始记载于《草本便方》，古书道，藤茶叶甘温消渴，诸气鼓胀月瘕活，丹停气肿下盅毒，利便通肠代茶喝。现代研究表明，藤茶具有抗炎镇痛、抗氧化、抗肿瘤、保肝、增强免疫力等作用。一方面，藤茶总黄酮对酒精性肝损伤小鼠具有一定保护作用,其可能机制与增加肝组织抗氧化酶的活性、降低脂质过氧化水平、降低肝组织IL-4、IL-6、TNF-α等炎症因子水平有关。两一方面，研究表明，藤茶黄酮能降低大鼠血脂，加强集体的抗氧化能力，同时改善高脂大鼠模型肝细胞的受损程度，从而促进肝再生。
桑黄（Phellinus igniarius (L. ex Fr.) Quel.），为多孔菌科真菌火木层孔菌的子实体，具有抗肿瘤、降压、降血糖、抗炎、护肝等药理作用。桑黄共有9种化合物,经筛选得到3个化合物、27个靶点、30个基因,与护肝相关的有MAOB、XDH、ALDH2，SMAD3、ALDH3A1、CYP2A6等22个关键基因。主要通过组氨酸代谢（histidinemetabolism）、酪氨酸代谢（tyrosine metabolism）等7条通路发挥作用。桑黄是我国传统中药，在众多护肝经方中出现，常配伍菟丝子、续断、龟板等，疗效显著。
历代文献和现代医学记载葛根对“酒病”的治疗有广泛应用。炒黄芩长在清燥中焦之湿热，归肺、胆、脾、大肠、小肠经。”明代《本草纲目》载“葛花、枳椇消酒积。”枳椇子长于止烦除渴，醒酒安神，归心、脾、肺经。三者配伍可清泄里热，解肌散邪，除酒毒酒热，达到解酒的功效。同时佐使炙甘草，助葛根升发，且调和诸药。炒白芍利于柔肝止痛，敛阴止汗，归肝、脾经。姜黄长于破血行气、通经止痛，归心、脾、胃经。丹参用于活血祛瘀、通经止痛，归心、肝经。三者配伍能养阴润燥，活血化淤，主治肝肾虚弱等饮酒后副作用，达到保肝护肝的作用。现代医学也进一步证实其有良好的降血脂、抗脂肪肝、保肝、促进肝再生的作用。熟地黄长于大滋肾水，养阴润燥，归肝、肾经。玄参长于滋肾养阴，解毒散结，归脾、肾、胃经。佐使五味子，既能收敛浮火，又可补肾阴。熟地黄为君，玄参长于滋肾养阴，填精益髓，解毒散结，二者协调滋阴补肾，契合肾生骨髓，髓生肝促进肝再生的治则。通过文献检索和前期药理药效学实验，我们将以上的中药进行配伍，多靶点、多环节来用于解酒保肝、促进肝再生。
本发明提供的中药组合物安全性和药效实验可降低动物模型血清ALT和AST水平，降低肝组织TG和MDA含量，并逆转肝损伤过程中的炎症反应和细胞衰老，从而起到解酒保肝、促进肝再生的作用。
3．有益效果
相比于现有技术，本发明的优点在于：
（1）本发明提供的解酒保肝、促进肝再生的中药组合物配方中的原料都属于既食既药的中药，安全性实验证明最大耐受剂量为临床成人用量的173.3倍，安全性良好。
（2）本发明提供的解酒保肝、促进肝再生的中药组合物对酒精刺激的体内慢性肝损伤具有显著防护作用。
（3）本发明通过葛根提取物，炒白芍提取物，熟地黄提取物，丹参提取物，姜黄提取物份，五味子提取物，玄参提取物，炙甘草提取物，炒黄芩提取物，枳椇子提取物，藤茶提取物份，桑黄提取物组合配伍，可通过减轻氧化应激导致的脂质过氧化作用发挥防治酒精性肝损伤效应；同时，本组合物可增强乙醇脱氢酶活性，加速体内乙醇代谢，提高抗氧化酶GSH等含量，减轻酒精导致的脂质过氧化，改善脂质代谢和血脂水平，缓解酒精诱导的脂质代谢紊乱。
（4）本发明通过将葛根提取物，炒白芍提取物，熟地黄提取物，丹参提取物，姜黄提取物份，五味子提取物，玄参提取物，炙甘草提取物，炒黄芩提取物，枳椇子提取物，藤茶提取物份，桑黄提取物进行组合配伍，发挥抗ALD和促进肝再生的疗效。
附图说明
图1为本发明药效学实验HE染色显示肝组织的结果示意图；
图2为本发明药效学实验免疫荧光染色显示的结果示意图；
图3为本发明药效学实验各组小鼠血清ALT、AST及肝组织TG、MDA水平图；
图4为本发明药效学实验各组小鼠肝组织的SIRT1,Cyclin E1, VDCA1和NF-κB的蛋白表达示意图及水平图。
具体实施方式
下面将结合实施例，对本发明实施例中的技术方案进行清楚、完整地描述：
实施例1：制成片剂
将葛根提取物4份，炒白芍提取物2份，熟地黄提取物4份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2 份，桑黄提取物3份，加入淀粉、羧甲基纤维素，混合均匀，制粒，压片，包衣，制成片剂。
实施例2：制成片剂
将葛根提取物5份，炒白芍提取物2份，熟地黄提取物4份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，加入淀粉、羧甲基纤维素，混合均匀，制粒，压片，包衣，制成片剂。
实施例3：制成片剂
将葛根提取物4份，炒白芍提取物2份，熟地黄提取物4份，丹参提取物2份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，加入淀粉、羧甲基纤维素，混合均匀，制粒，压片，包衣，制成片剂。
实施例4：制成片剂
将葛根提取物4份，炒白芍提取物2份，熟地黄提取物4份，丹参提取物3份，姜黄提取物3份，五味子提取物2份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，加入淀粉、羧甲基纤维素，混合均匀，制粒，压片，包衣，制成片剂。
实施例5：制成片剂
将葛根提取物5份，炒白芍提取物2份，熟地黄提取物5份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，加入淀粉、羧甲基纤维素，混合均匀，制粒，压片，包衣，制成片剂。
实施例6：制成硬胶囊剂
将葛根提取物4份，炒白芍提取物2份，熟地黄提取物5份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，混合均匀，粉碎，过三号筛，制成颗粒，70-90℃条件下干燥，整粒，填充明胶硬胶囊，制成硬胶囊剂，每颗胶囊内容物0.5g。
实施例7：制成硬胶囊剂
将葛根提取物6份，炒白芍提取物2份，熟地黄提取物5份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物3份，炒黄芩提取物3份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份，混合均匀，粉碎，过三号筛，制成颗粒，70-90℃条件下干燥，整粒，填充明胶硬胶囊，制成硬胶囊剂，每颗胶囊内容物0.5g。
本发明的配方命名为复方肝泰生，并进行一系列的安全性和药效性实验，结果如下：
1.安全性实验
目的：观察小鼠给子受试品后活动情况和死亡情況，确定配方安全性。受试样品：促进肝再生的中药组方干浸膏方法：常温下配制受试品最高浓度为4.44 g/ml，以最大体积0.8 ml经口灌胃给予小鼠2次（间隔8 h），连续观察7天，详细记录小鼠反应情况。灌胃后活动情況和死亡情况，确定配方安全性。结果：每次给受试品后,小鼠活动正常，且在灌胃后无小鼠出现死亡，无其他异常情况。连续观察14天后,无小鼠出现死亡和其他异常情况，体重持续增长。结论：临床成人用量每日10 g（生药量），按成人体重60 kg，小鼠体重20 g计算，小鼠24 h内口服受试品最大耐受剂量为4.44*0.8*2 /0.02=355.2 g/kg，为临床成人用量的355.2 / 12.3 /(10/60)=173.3倍，安全性达到实际无毒类范畴。( 12.3为人和小鼠用药的换算系数，实际用量依据具体实验数量而定)
2.药效学实验
采用经典的Lieber-DeCarli液体酒精饲料(含酒精5％)和无酒精的液体饲料，定量饲喂小鼠(保持摄入的热量一致)，复制酒精性肝损伤模型。
野生型C57BL/6小鼠按体重随机分为4组，每组10只，分别为无酒精的液体饲料正常对照组、液体酒精饲料模型组、低剂量中药组合物（复方肝泰生300mg/kg)组、高剂量中药组合物（复方肝泰生600mg/kg）组。正常对照组给予无酒***体饲料，其余组给予同热量同体积含酒精的Lieber-DeCarli液体饲料，饲养8周，结束后禁食12h，眼眶取血，采用全自动生化分析仪测定血清谷丙转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。分离肝组织做病理切片(HE染色)及取肝组织匀浆上清检测甘油三酯(TG)、丙二醛(MDA)水平，并采用Western-blot法检测肝再生及肝损伤相关指标，结果用GraphPad统计并进行单因素方差分析，所有数据以(x±s)表示。
结果：在酒精肝损伤造模成功的前提下，高低剂量的中药组合物（复方肝泰生）组血清ALT和AST、肝组织TG和MDA水平显著低于酒精模型对照组，且呈剂量依赖性（图3）。HE染色显示，高低剂量的中药组合物（复方肝泰生）组的肝组织异常的脂质沉积和炎症反应较模型组显著改善（图1）。
Western-blot显示高低剂量的中药组合物（复方肝泰生）组肝组织中SIRT1、cyclin E1表达显著增加，VDCA1和NF-κB表达显著降低（图4）。此外免疫荧光染色显示，受试样品组NF-κB入核显著降低（图2）。以上结果表明复方肝泰生可显著改善慢性酒精导致的细胞衰老和肝再生障碍。
Claims (3)
1.一种用于解酒保肝、促进肝再生的中药组合物，其特征在于：所述中药组合物由以下重量份数配比的组分制成：葛根提取物4份，炒白芍提取物2份，熟地黄提取物5份，丹参提取物3份，姜黄提取物1份，五味子提取物1份，玄参提取物3份，炙甘草提取物2份，炒黄芩提取物2份，枳椇子提取物3份，藤茶提取物2份，桑黄提取物3份；
所述方法包括如下步骤，先将所述中药组合物中的葛根、炒白芍、熟地黄、丹参，姜黄，五味子，玄参，炙甘草，炒黄芩，枳椇子，藤茶及桑黄的提取物通过以下方法获得：
1)将葛根实体破碎，加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到葛根提取物粉末；
2)将炒白芍破碎，加入3-7倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炒白芍提取物粉末；
3)将熟地黄加入4-6倍量水煎煮4-6次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到熟地黄提取物粉末；
4)将丹参加入3-7倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；浸膏冷冻真空干燥，粉碎过六号筛，得到丹参提取物粉末；
5)将姜黄加入1-4倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到姜黄提取物粉末；
6)将五味子加入1-4倍量水煎煮1-2次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到五味子提取物粉末；
7)将玄参加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到玄参提取物粉末；
8)将炙甘草加入1-4倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炙甘草提取物粉末；
9)将炒黄芩加入3-8倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到炒黄芩提取物粉末；
10)将枳椇子加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到枳椇子提取物粉末；
11)将藤茶加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到藤茶提取物粉末；
12)将桑黄加入5-10倍量水煎煮2-3次，每次1小时，去渣取药汁合并滤液，滤液在70-90℃条件下减压浓缩，得到浸膏；将浸膏冷冻真空干燥，粉碎过六号筛，得到桑黄提取物粉末；
13)最后将葛根，炒白芍，熟地黄，丹参，姜黄，五味子，玄参，炙甘草，炒黄芩，枳椇子，藤茶，桑黄的这12种提取物的浸膏干粉按照一定比例混合。
2.一种用于解酒保肝、促进肝再生的中药组合物制剂，其特征在于：所述中药制剂由权利要求1所述的中药组合物制成。
3.根据权利要求2所述的一种用于解酒保肝、促进肝再生的中药组合物制剂，其特征在于：所述中药制剂为片剂、粉剂、颗粒剂、冲剂、胶囊剂、膏剂、丸剂或液体制剂。
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Inventor after: Qiu Ping Inventor after: Zhu Guanghan Inventor after: Shi Weixi Inventor after: Yan Honghao Inventor after: Zhang Lu Inventor after: Cao Chunlu Inventor after: Wang Shihui Inventor after: Yao Jiapeng Inventor after: Lou Kezao Inventor after: Ma Qing Inventor after: Du Wangzhe Inventor after: Xu Ke Inventor after: Luo Qihan Inventor after: Jiang He Inventor after: Gu Nanjun Inventor after: Huang Shuo Inventor before: Lou Kezao Inventor before: Zhu Guanghan Inventor before: Shi Weixi Inventor before: Yan Honghao Inventor before: Zhang Lu Inventor before: Cao Chunlu Inventor before: Wang Shihui Inventor before: Yao Jiapeng Inventor before: Qiu Ping Inventor before: Ma Qing Inventor before: Du Wangzhe Inventor before: Xu Ke Inventor before: Luo Qihan Inventor before: Jiang He Inventor before: Gu Nanjun Inventor before: Huang Shuo |