CN108310080A - 黑果腺肋花楸花色苷在制备抗肝纤维化药品或保健品中的应用 - Google Patents
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Abstract
本发明涉及黑果腺肋花楸花色苷在制备抗肝纤维化药品或保健品中的应用。黑果腺肋花楸花色苷可以单用或与其他具有抗肝纤维化作用的药物组合，按常规制剂的制备方法，制备成适合口服的任何剂型的药品或保健品。黑果腺肋花楸花色苷具有很强的预防肝纤维化的能力。
Description
技术领域
本发明属于保健食品和药物领域，具体地涉及黑果腺肋花楸花色苷在制备具有治疗和预防肝纤维化的药品或保健品中的应用。
背景技术
肝纤维化(fibrosis of liver)是多种原因引起的慢性肝损害所致的病理改变，表现为肝内细胞外间质成分过度异常地沉积，并影响肝脏的功能，是慢性肝病发展到肝硬化必经之阶段。现认为肝纤维化尚有逆转至正常的可能，而肝硬化则否。目前研究的重点放在分子与分子、分子与细胞及细胞与细胞间的相互作用方面。在诊治方面虽然有一些进展，但仍缺乏确定有效的药物。
肝纤维化保健品能在一定程度上预防肝纤维化，促进身体健康，减少肝损伤。目前抗肝纤维化药物可以分为传统中药和现代药物两大类。例如安络化纤丸、复方鳖甲软肝片等药物，能软坚散结，化瘀解毒，益气养血，疏通肝血流、清除肝内淤积、降低门静脉压、防止出血、促使肝纤维降解吸收和阻断肝内纤维增生，预防、阻断和逆转肝纤维化。而化学合成的药物，其成本大，毒性大，副作用强，会影响内分泌功能和消化道反应等副作用。因此，从天然植物中寻找具有生物活性的、预防肝纤维化作用的物质应用于医药、食品和保健品中具有重要的意义。
黑果腺肋花楸(Aornia mealnocarpa Elliot)又称野樱莓、不老莓，是集食用、药用、园林和生态价值于一身的珍贵树种。原产于北美东北部，为近年来辽宁省主要引进经济树种。
发明内容
本发明以寻求新型高效的预防肝纤维化药品或保健品的开发利用为目的，提出了黑果腺肋花楸花色苷在制备治疗和预防肝纤维化药品或保健品中的应用。
本发明采用黑果腺肋花楸花色苷作为原料，单用或与其他具有抗肝纤维化作用的药物组合，按常规制剂的制备方法，制备成适合口服的任何剂型的抗肝纤维化药品或保健品，如口服液、片剂、胶囊或颗粒剂，或其它针剂剂型。
优选的，所述的黑果腺肋花楸花色苷是醇提化合物。
优选的，所述的黑果腺肋花楸花色苷的制备方法包括如下步骤：60-70℃下，将黑果腺肋花楸果实置于乙醇中浸泡1-2h后，超声30min，连续超声2次，产物经大孔吸附树脂纯化后，所得提取液旋转蒸发，浓缩，真空冷冻干燥，得黑果腺肋花楸花色苷。
更优选的，所述的乙醇为体积百分浓度为60％的乙醇。所述的大孔吸附树脂为大孔吸附树脂AB-8。
本发明的有益效果是：本发明采用的是黑果腺肋花楸花色苷，以四氯化碳诱导的方式诱导小鼠快速肝纤维化，通过肝纤维化模型小鼠体内实验得出，黑果腺肋花楸花色苷可以降低肝纤维化小鼠的脏器指数，以及肝纤维化标志性酶即血清和肝脏组织中谷草转氨酶和谷丙转氨酶含量下降，并维持小鼠肝组织结构完整清晰，肝细胞大小均匀，无变性、坏死、细胞核大而圆，多居于细胞中央。黑果腺肋花楸含有极为丰富的花色苷物质，具有较强的抗氧化活性，较强的抗炎功效，以及一定的治疗血管疾病的功效，还具有抗癌和保胃的功能，本发明经研究发现，黑果腺肋花楸花色苷具有较强的抗肝纤维化的功效，是开发新型高效的天然抗肝纤维化药物或保健品的一个好选择。
附图说明
图1是各组小鼠肝系数的比较结果。
图2A是各组小鼠血清中谷草转氨酶(AST/GOT)含量比较结果。
图2B是各组小鼠血清中谷丙转氨酶(ALT/GPT)含量比较结果。
图3A是各组小鼠肝组织中谷草转氨酶(AST/GOT)含量比较结果。
图3B是各组小鼠肝组织中谷丙转氨酶(ALT/GPT)含量比较结果。
图4是各组小鼠肝组织HE染色情况比较图。
具体实施方式
实施例1
(一)制备方法
黑果腺肋花楸花色苷提取物的制备：称取黑果腺肋花楸果实100g，用60％的乙醇在60-70℃浸泡1h，放入超声清洗器中超声30min，连续2次，经大孔吸附树脂AB-8纯化后，再用旋转蒸发仪浓缩提取液，经过真空冷冻干燥，得黑果腺肋花楸花色苷干粉。
经检测，花色苷干粉主要含有47.6％的矢车菊素阿拉伯糖苷(cyanidin 3-arabinoside)和46.4％的矢车菊素半乳糖苷(cyanidin 3-galactoside)成分。
(二)药效试验
动物饲养及处理：实验动物饲养1周以适应本实验室条件，分笼饲养，室内无菌消毒，每两天换一次垫料。控制实验室光照，12h明-12h暗，室内温度为20±1℃，湿度为55±10％。
实验动物分组：雄性小鼠50只，体重30±2g，随机分为5组如下，每组10只。
(1)正常对照组(C)；
(2)肝纤维化模型组(M)；
(3)阳性对照组(E)；
(4)给药组：黑果腺肋花楸花色苷组40mg/L(H)；
(5)给药组：黑果腺肋花楸花色苷组20mg/L(L)
正常对照组10只小鼠皮下注射生理盐水，其余小鼠注射20％四氯化碳橄榄油混合液(150mg/kg)，每日一次，持续8周。造模的同时进行给药，除正常对照组、肝纤维化模型组小鼠给予生理盐水饮水外，给药组小鼠分别给予花楸花色苷20mg/L和40mg/L，每一周称量并记录小鼠体重。实验8周时，进行检测。
1.采血后处死小鼠，收集肝组织并称重，计算肝系数。肝系数的比较结果见图1。
如图1所示，肝纤维化模型组小鼠肝系数明显高于正常组(8.98vs 10.96,p<0.01)，黑果腺肋花楸花色苷给药能干预肝纤维化引起的肝系数升高，相较于肝纤维化模型组，给药组H可以显著抑制小鼠肝系数的升高(10.70,p<0.01)。实验结果说明黑果腺肋花楸花色苷对于小鼠肝纤维化过程中肝增大具有显著的干预作用且呈剂量相关性。
2.检测血清和肝组织内谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量，结果见图2A，图2B，图3A和图3B。
如图2A和图2B所示，肝纤维化模型组小鼠血清的谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量明显高于正常组，黑果腺肋花楸花色苷给药可以降低肝纤维化模型小鼠血清的谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量，且其对含量的降低具有显著性(给药组L为p<0.05，给药组H为p<0.001)。实验结果说明黑果腺肋花楸花色苷对小鼠肝纤维化过程中血清中谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量具有一定的干预作用，并且呈剂量相关性。
如图3A和图3B所示，肝纤维化模型组小鼠肝组织中的谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量明显高于正常组，黑果腺肋花楸花色苷给药可以降低肝纤维化模型小鼠肝组织中的谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量，且其对含量的降低具有显著性(给药组L为p<0.05，给药组H为p<0.001)。实验结果说明黑果腺肋花楸花色苷对小鼠肝纤维化过程中肝组织中谷草转氨酶(AST/GOT)和谷丙转氨酶(ALT/GPT)的含量具有一定的干预作用，并且呈剂量相关性。
3.HE染色观察肝纤维化小鼠肝组织的结构和形态，结果见图4。
如图4所示，HE切片结果显示，空白对照组小鼠肝脏标本肝小叶结构正常，肝细胞索排列规则，肝细胞无变性、坏死，未见增生纤维组织。肝纤维化模型组小鼠肝细胞排列欠规则，有部分肝细胞坏死、脂肪变性及气球样变，汇管区纤维化扩大，窦周可见少许纤维组织增生，汇管区可见线状短纤维间隔形成。黑果腺肋花楸花色苷给药组,肝细胞变性无坏死，汇管区纤维沉积较模型组明显减少，略有炎症浸润，其中高剂量给药组更具显著性的影响。
Claims (7)
1.黑果腺肋花楸花色苷在制备抗肝纤维化药品或保健品中的应用，其特征在于，黑果腺肋花楸花色苷单用或与其他具有抗肝纤维化作用的药物组合制备抗肝纤维化药品或保健品。
2.根据权利要求1所述的应用，其特征在于：所述的黑果腺肋花楸花色苷是醇提化合物。
3.根据权利要求2所述的应用，其特征在于：所述的黑果腺肋花楸花色苷的制备方法包括如下步骤：60-70℃下，将黑果腺肋花楸果实置于乙醇中浸泡1-2h后，超声30min，连续2次，产物经大孔吸附树脂纯化后，所得提取液旋转蒸发，浓缩，真空冷冻干燥，得黑果腺肋花楸花色苷。
4.根据权利要求3所述的应用，其特征在于，所述的乙醇为体积百分浓度为60％的乙醇。
5.根据权利要求3所述的应用，其特征在于，所述的大孔吸附树脂为大孔吸附树脂AB-8。
6.根据权利要求1或2所述的应用，其特征在于，所述的黑果腺肋花楸花色苷的剂量为0.3-3mg/d。
7.根据权利要求1或2所述的应用，其特征在于，所述的药品或保健品制成口服液、片剂、胶囊、颗粒剂针剂剂型。
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