CN107875180A - 辅酶q10与月见草油制备预防骨质疏松的保健食品配方及生产工艺 - Google Patents
辅酶q10与月见草油制备预防骨质疏松的保健食品配方及生产工艺 Download PDFInfo
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Abstract
本发明涉及用辅酶Q10与月见草油组成的防治骨质疏松症的保健食品与药品，研究发现，把辅酶Q10与月见草油一起组成复合制剂，其抗氧化，促进成骨细胞形成的作用明显增强，抗骨质疏松的作用特别明显，对骨质疏松有更好的预防作用；进一步研究还发现，把辅酶Q10溶解于其中，当辅酶Q10在月见草油中的含量达到3～25％时，即每毫升月见草油中辅酶Q10含量为0.03～0.25g时，就可以发挥预防骨质疏松的作用。本发明辅酶Q10溶解在月见草油后，可以按照软胶囊的生产工艺，添加药剂学上的辅料，制备为软胶囊，作为临床预防骨质疏松的保健食品及药品。
Description
技术领域
本发明涉及用辅酶Q10与月见草油组成的防治骨质疏松症的保健食品与药品，该药物组合物对妇女绝经期后骨质疏松，老年性骨质疏松具有较好的预防及治疗作用。
背景技术
原发性骨质疏松症是一种以低骨量和骨组织微结构破坏为特征，导致骨骼脆性增加和易发生骨折的全身性疾病，可分为绝经后骨质疏松和老年性骨质疏松两型。绝经期后骨质疏松是60岁以上妇女的常见病，多发病，主要是由于雌激素缺乏所致，常发生于绝经后数年内的女性，其中多数患者的骨转换率增高，旧骨大量迅速丢失，新骨形成加快，骨的新旧转换速率加速，但由于骨丢失超过了骨形成，骨量以丢失为主，形成了骨质疏松，故该型的骨质疏松也称为高转换型骨质疏松。
氧化应激作为骨质疏松的一个危险因子已逐渐受到重视，该学说认为，氧化应激是由于活性氧自由基过剩或抗氧剂缺乏引起的氧化还原系统失衡状态，可导致许多组织损伤，最终引起机体的衰老和疾病，氧化应激直接或间接地影响骨吸收与骨形成的过程，与氧化应激损伤相关的早衰鼠模型则显示骨质疏松症状，骨质疏松病理机制的焦点逐渐从“以雌激素缺乏为中心”转向“以衰老和氧化应激为中心”。研究表明伴随着衰老进程，机体的氧化蛋白产物(AOPP)和脂质过氧化产物丙二醛(MDA)增加，超氧化物歧化酶(SOD)活性降低，同时，骨密度、松质骨体积、骨小梁厚度、骨形成率及矿化沉积率均减小，可见，氧化应激在衰老导致骨质疏松的发生中起着重要作用。对氧化应激影响骨代谢信号通路的研究证明，细胞内的多条信号通路相互协调，共同控制着骨代谢的平衡，如RANKL介导的信号通路调控破骨细胞的分化和骨基质的降解；胰岛素信号通路的胰岛素样生长因子-1(IGF-1)可提高碱性磷酸酶(ALP)活性，促进胶原的合成与分泌，减少胶原降解，增强基质蛋白的附着，并调节骨钙素的转录后修饰与表达，促进成骨细胞增殖分化及矿化结节的形成；Wnt信号通路可调节成骨细胞的分化、骨基质的形成及矿化，还可调节OPG/RANKL的表达，间接调节破骨细胞的生成和骨吸收，影响骨重塑过程。
氧化应激与体内自由基的生成与清除密切相关。活性氧自由基(ROS)主要产生于线粒体。在正常生理条件下，ROS可被过氧化氢酶(CAT)、过氧化物酶(POD)及超氧化物歧化酶(SOD)分解，使自由基的生成与清除处于动态平衡，但在氧化应激、疾病和衰老等情况下，线粒体内产生过量的ROS，同时，抗氧化系统的损伤使ROS清除变慢，未被分解或中和的ROS会攻击线粒体，损伤线粒体内膜的呼吸链，使线粒体中的酶活性降低；促进线粒体DNA突变；改变线粒体膜的通透性，使线粒体膜电位下降；诱导MPTP孔开放并促进凋亡诱导因子和细胞色素C的释放，通过激活caspase级联反应或其他凋亡相关蛋白引发细胞凋亡；ROS还会破坏细胞内的氧化还原平衡，导致细胞内蛋白质、脂质及核酸等多种成分发生氧化损伤。在调控骨代谢的信号通路中，MAPK/ERK监控AMP与ADP水平，调节细胞的生长代谢及能量平衡，雌激素与其受体结合调控相关转录因子的表达，PI3K/Akt磷酸化FoxOs，以及Nrf2.Keapl调控二相酶防御系统等均与线粒体结构、功能以及动态变化相关，共同参与调节成骨细胞与破骨细胞的分化及功能。因而，从线粒体角度探究氧化应激导致骨丢失的分子机制和药物作用新靶点具有重要意义。
辅酶Q10也称泛醌，它在人体内可以自动合成。辅酶Q10在人体大多数组织(如心脏、骨骼肌、大脑、肾脏和肝脏)的ATP形成中起着至关重要的作用，同时它具有抗氧化功能。20世纪60年代中期开始用辅酶Q10辅助治疗心脏病，后来辅酶Q10的用途逐渐拓展，目前已广泛用于治疗心绞痛、心肌衰竭等心血管疾病，以及辅助治疗高血压和神经系统疾病。辅酶Q10是一种强抗氧化剂，它可以通过抑制脂质的过氧化、终止蛋白质的氧化等方式起到抗氧化作用，辅酶Q10能阻止脂和蛋白质的过氧化，清除自由基，明显降低线粒体氧耗，减少细胞三磷酸腺苷消耗，维持细胞线粒体结构的完整性，防止细胞水肿，细胞膜破裂，以及线粒体溶解与肌纤维的紊乱性排列，辅酶Q10还能通过氧化还原性结构的变换，加强细胞内膜抵抗外界氧化因子的损害，是人体内唯一的一种天然存在的、又能在人体中再生的脂溶性抗氧化剂。迄今为止，没有发现辅酶Q10的明显毒副作用。
小麦胚芽油经由小麦胚芽经过压榨或浸出工艺制取的一种油脂，它集中了小麦的营养精华，富含油酸、亚油酸、亚麻酸、甘八碳醇及多种生理活性组分，维生素E含量为植物油之冠，已被公认为一种具有营养保健作用的功能性油脂，具有很高的营养价值，被营养学家誉为天然维生素E的仓库,维生素E，α、β、γ、δ四种类型全具备，生理活性效能是合成维生素E的30倍，易被人体吸收。在体内防止氧化脂质的生成，保护细胞膜，抑制自由基，促进人体新陈代谢，延缓机体衰老，改善肝脏功能，降低胆固醇，提高免疫力，是为数不多的真正应用于人类抗衰老的抗氧化剂。能预防和辅助治疗一些中老年疾病，如心脏病、脑中风、心肌梗塞、高血脂、动脉硬化、肺气肿、更年期障碍、贫血、肌肉营养不良等。小麦胚芽油中人体必需不饱和脂肪酸含量高达80％以上，仅亚油酸的含量就占到55％以上。不饱和脂肪酸能保持细胞膜的相对流动性，保证细胞的正常生理功能，使胆固醇酯化，降低血中胆固醇及甘油三酯，降低血液粘稠度，改善血液微循环，提高脑细胞的活性，增强思维能力和记忆力。
本项目组研究发现，小麦胚芽油对骨质疏松有明显的预防作用，该用途已经申报了国家发明专利，专利号：200610110559.5，并已经获得国家知识产权局的授权(授权日：2010.05.12，专利证书号：第617477号)；进一步研究发现，把辅酶Q10与小麦胚芽油一起组成复合制剂，其抗氧化，促进成骨细胞形成的作用明显增强，抗骨质疏松的作用特别明显，对骨质疏松有更好的预防作用，实验观察表明，当辅酶Q10溶解于小麦胚芽油中，达到20mg/ml时，对去卵巢大鼠导致的骨质疏松，对环磷酰胺导致的大鼠骨质疏松，均有非常明显的预防作用。
本项目组进一步研究还发现，分别用芝麻油，月见草油，大豆油，花生油，紫苏子油，核桃油代替小麦胚芽油，把辅酶Q10溶解于其中，当辅酶Q10在植物油中的含量达到3～25％时，即每毫升植物油中辅酶Q10含量为0.03～0.25g时，也可以发挥预防骨质疏松的作用。
为了减少辅酶Q10的添加量，本项目组进一步研究还发现，在辅酶Q10溶解在植物油中后，添加人参提取物或人参皂苷，可以到达同样的作用，其中，人参皂苷的添加量为每毫升植物油中0.06～0.5g时，辅酶Q10的含量为人参皂苷的二分之一配伍，也可以达到预防骨质疏松的作用。
本项目组进一步研究还发现，在辅酶Q10溶解在植物油中后，再添加甘草提取物甘草酸或甘草次酸，也可以到达同样的作用，其中，甘草次酸的添加量为每毫升植物油中0.06～0.5g时，辅酶Q10的含量为甘草次酸的二分之一配伍，也可以达到预防骨质疏松的作用。
本发明辅酶Q10溶解在植物油中后，可以按照软胶囊的生产工艺，添加药剂学上的辅料，制备为软胶囊，作为临床预防骨质疏松的保健食品及药品应用。
本发明辅酶Q10溶解在植物油中后，还可以加上药剂学方面的辅料，制成含片，胶囊，片剂，颗粒剂，冲剂，口服液，喷雾剂，注射剂，作为临床预防骨质疏松的保健食品及药品应用。
具体实施方式：实施例一
为证实辅酶Q10植物油组合物具有抗骨质疏松作用，本实验以环磷酰胺致大鼠骨丢失建立骨质疏松模型，通过骨形态计量学检测等研究方法，观察辅酶Q10复方制剂是否可以预防化疗药物导致的大鼠骨质疏松。现报告如下：
1材料和方法
1.1材料
实验动物及分组108只3月龄SPF级SD大鼠，雄性，按随机区组法即配伍分组法原则，先将动物分别按体重区分为基干个区组，然后按名区组通过Excel软件的随机函数Rand平均分配到11组：①空白组：灌胃给予蒸馏水5ml/kg/d；②模型组：灌胃给予环磷酰胺2ml/kg/d和生理盐水3ml/kg/d；给药组：分组给药见表1：
表2实验动物分组及给药剂量
1.2药物制备：
环磷酰胺：取环磷酰胺1支0.2g，用蒸馏水溶解定容至80ml，现配现用(2h内用完)。大鼠按每天5mg/kg给药
阿仑膦酸钠：取阿仑膦酸钠半片(5mg)，研钵研细后，蒸馏水溶解定容至15ml。---1mg/kg/d
辅酶Q10复方1组：取辅酶Q10 3g，小麦胚芽油100ml；混匀超声10min。
辅酶Q10复方2组：取辅酶Q10 3g，芝麻油100ml；混匀超声10min。
辅酶Q10复方3组：取辅酶Q10 3g，月见草油100ml；混匀超声10min。
辅酶Q10复方4组：取辅酶Q10 3g，花生油100ml；混匀超声10min。
辅酶Q10复方5组：取辅酶Q10 3g，紫苏子油100ml；混匀超声10min。
辅酶Q10复方6组：取辅酶Q10 3g，核桃油100ml；混匀超声10min。
辅酶Q10复方7组：取辅酶Q10 1g，人参皂苷2g，芝麻油100ml；混匀超声10min。
辅酶Q10复方8组：取辅酶Q10 1g，甘草次酸2g，芝麻油100ml；混匀超声10min。
1.2方法
所有动物均灌胃给药，每天一次，连续给药15天，在取骨标本前14、13天和4、3天分别皮下注射四环素30mg/kg和钙黄绿素10mg/kg进行体骨内荧光标记。实验结束时，大鼠麻醉后心脏抽血处死，取血液做生化指标测定，并取左胫骨做骨组织形态计量学研究，取左股骨做骨密度扫描。。
骨组织形态计量取左侧胫骨，以慢速锯暴露骨髓腔，取其上段以10％福尔马林缓冲液固定24h、随后进行梯度酒精脱水，二甲苯透明以甲基丙烯酸甲酯进行不脱钙骨包埋。包埋剂硬化后以石膏打磨机打磨成合适的形状，随后用Leica 2155型硬组织切片机切成9μm厚片和5μm薄片。9μm厚片直接封片测量动态参数；5μm薄片行硝酸银染色，封片后测量静态参数，运用公式[6,7]计算有关参数。
胫骨上段测量范围为生长板远端1mm～4mm之间的松质骨区域。静态测量参数包括骨组织总面积(T.Ar)、骨小梁面积(Tb.Ar)、骨小梁周长(Tb.Pm)、破骨细胞个数(N.Oc)、破骨细胞贴壁长度(Oc.S)。动态测量参数包括单荧光周长(sL.Pm)、双荧光周长(dL.Pm)、双标记间隙(Ir.L.Wi)。参照相关公式，可以计算出计算参数：骨小梁面积百分数(％Tb.Ar)降低、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)、骨小梁分离度(Tb.Sp)、每毫米破骨细胞数目(Oc.N)、破骨细胞周长百分数(％Oc.Pm)、标记周长百分数(％L.Pm)、骨矿化形成率(MAR)、骨形成率(BFR/BV)、骨形成率(BFR/BS)、骨形成率(BFR/TV)。
2结果
本研究通过骨组织形态计量的测定及评价，单独应用环磷酰胺组大鼠静态测量参数包括骨组织总面积(T.Ar)、骨小梁面积(Tb.Ar)、骨小梁周长(Tb.Pm)明显减少，骨量明显丢失，呈现骨质疏松的典型表现。应用辅酶Q10复方预防组，各组大鼠骨量均有明显的增加作用，与环磷酰胺模型组相比，有显著性差异，与正常对照组相比，差异没有显著性。
研究结果说明，辅酶Q10组成的八个复方，均有明显的预防骨质疏松作用。
实施例二
根据本发明提出的辅酶Q10植物油组合物的制备工艺，把辅酶Q10溶解于植物油中，再按软胶囊的生产工艺，制备出供临床应用于抗骨质疏松功效的保健食品或药品软胶囊，再把这种软胶囊进行抗骨质疏松的药效学实验，通过以大鼠去卵巢建立骨质疏松模型，采用骨形态剂量学方法进行评价，结果表明，用本发明思路制备的辅酶Q10组合物复方软胶囊，具有十分明显的预防骨质疏松作用。
进一步研究还证明，本发明辅酶Q10溶解在植物油中后，还可以加上药剂学方面的辅料，制成含片，胶囊，片剂，颗粒剂，冲剂，口服液，喷雾剂，注射剂，这些剂型的保健食品或药品，作为临床预防骨质疏松的应用也有非常明显的作用。
Claims (4)
1.一种含月见草油和辅酶Q10的预防骨质疏松的保健食品和药品的软胶囊,其特征是每100毫升月见草油中加入3～25克辅酶Q10，其生产工艺是先把辅酶Q10溶解在月见草油中，然后，按照软胶囊的生产工艺，添加药剂学上的辅料，制备为软胶囊，即可。
2.如权利要求1所述的一种含月见草油和辅酶Q10的预防骨质疏松的保健食品和药品的软胶囊，其特征是该组合物中可以含有人参皂苷。
3.如权利要求1所述的一种含月见草油和辅酶Q10的预防骨质疏松的保健食品和药品的软胶囊,其特征是该组合物中可以含有甘草酸、甘草次酸。
4.如权利要求1、2、3所述的一种含月见草油和辅酶Q10的预防骨质疏松的保健食品和药品的软胶囊，其特征是该组合物可以加上药剂学方面的辅料，制成含片，胶囊，片剂，颗粒剂，冲剂，口服液，喷雾剂，注射剂。
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