CN113521195B - 一种用于酒精使用障碍的药食同源组合物及应用 - Google Patents
一种用于酒精使用障碍的药食同源组合物及应用 Download PDFInfo
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- CN113521195B CN113521195B CN202110868997.2A CN202110868997A CN113521195B CN 113521195 B CN113521195 B CN 113521195B CN 202110868997 A CN202110868997 A CN 202110868997A CN 113521195 B CN113521195 B CN 113521195B
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Abstract
本发明公开了一种用于酒精使用障碍的药食同源组合物及其应用,属于保健食品技术领域。本发明的组合物由石斛、黄精和葛根配伍组成。本发明的组合物对小鼠酒精性肝损伤有显著的保护作用,包括对肝脏细胞形态、结构的改善,对肝功能以及肝脏脂代谢能力的改善。另一方面,本发明的组合物能改善酒精戒断后的抑郁、焦虑样反应,降低酒精戒断引起的促肾上腺皮质激素释放激素、促肾上腺皮质激素和皮质酮的升高;抑制酒精戒断引起的单胺类递质多巴胺和去甲肾上腺素的减少。
Description
技术领域
本发明属于保健食品技术领域,具体涉及一种用于酒精使用障碍的药食同源组合物及其应用,包括对酒精性肝损伤的保护作用和对酒精成瘾后的戒断反应的新应用。
背景技术
过量或长期饮酒会引起中毒、成瘾和戒断症状等酒精使用障碍综合症。酒精的安全饮用量为零,世界卫生组织也早把酒精列为1级致癌物质。而嗜酒者停止饮酒后会产生强烈的戒断症状,重新饮酒后会产生更严重的戒断症状。
酒精被吸收后大部分在肝脏中代谢,只有约5%-10%酒精没有氧化分解,直接从尿中排出或经肺从呼吸道呼出或经皮肤汗腺排除。因此酒精对肝有直接的毒性作用,干扰脂类、糖类和蛋白质等营养物质的正常代谢,同时影响肝脏解毒功能。长期过量饮酒则会造成脂肪肝、酒精性肝炎、肝硬化等酒精性肝病。
除了肝脏功能受损外,酒精对中枢神经***也有严重的危害,而且长期饮酒会造成酒精性依赖、成瘾,当停止饮酒后会出现戒断症状,随依赖程度的加深及戒断次数的增多,患者可能出现抑郁、焦虑样症状。其机制涉及酒精造成单胺类神经递质代谢异常,下丘脑 -垂体-肾上腺(HPA)轴亢进等神经***病变,这也是酒精依赖患者反复戒断-复饮的原因之一。
临床上对酒精使用障碍的最常用治疗药物是双硫仑,它能够抑制肝细胞乙醛脱氢酶 (ALDH)活性,阻止酒精代谢,使酒精代谢停留在乙醛阶段,而积聚过多的乙醛可引起人体诸多的不适症状,因此对酒精产生暂时性厌恶,达到戒酒目的。但长期服用,累积的乙醛对消化***、心血管***和神经***都有毒害作用,严重者还能引起肝坏死。另外,纳曲酮(Naltrexone)被用于治疗急性酒精中毒,它是一种阿片类受体拮抗剂,容易产生依赖性。
我国传统本草书籍最早记载的具有解酒功效的中药是葛根(Pueraria lobata(Willd.) Ohwi),申请人前期也发现葛根对小鼠慢性酒精依赖性肝损伤有明显保护作用(Biomedicine&Pharmacotherapy 2020,127:110163)。为了更好地改善酒精引起的肝损伤及酒精戒断反应,申请人以具有滋补作用的石斛和黄精为主药,配伍葛根,以增强其补脾益气、醒酒保肝作用,并通过酒精成瘾和戒断的动物实验进一步研究本组合物的功效,扩展其应用价值。
发明内容
鉴于此,本发明的目的是提供了一种药食同源组合物及其在改善酒精性肝损伤和酒精成瘾后的戒断反应中应用。
本发明目的是通过以下方式实现:
本发明提供一种药食同源组合物,由以下原料组成:石斛提取液9-12份,黄精提取液8-10份,葛根粉1-3份。
进一步地,基于以上技术方案,所述的石斛提取液是通过以下方法制备得到的:取含水量在75-90%的新鲜石斛,加水至新鲜石斛和水的质量比为1:1-1:3,于榨汁机中破碎5-15 分钟,过滤,收集滤液;向滤渣中加水至滤渣和水的质量比为1:0.8-1:15,在45-65℃条件下,超声15-25分钟,过滤,合并2次滤液,静置;然后在3000-5000r/min条件下,离心10-20分钟,收集上清液,浓缩,制成0.4-0.6g/ml石斛提取液,低温保存备用。
进一步地,基于以上技术方案,所述的黄精提取液是通过以下方法制备得到的:取干燥的黄精药材,加水至黄精和水的质量比为1:2-1:8,浸泡3-5h,然后转入破壁机内,破碎6-12分钟;在70-90℃的条件下水浴温煮1-4h,过滤,向滤液中加入乙醇,使滤液中乙醇终浓度为70-80%,充分搅拌后静置;然后在3000-5000r/min条件下离心10-20分钟,收集上清液,浓缩,制成0.3-0.5g/ml黄精提取液,低温保存备用。
本发明另一方面提供上述的药食同源组合物在改善酒精性肝损伤和/或酒精成瘾后的戒断反应中应用。
本发明再一方面提供上述的药食同源组合物在制备改善肝损伤和/或酒精成瘾后的戒断反应的食品或药物中应用。
进一步地,基于以上技术方案,所述的食品或药物能够抑制酒精引起的谷丙转氨酶和谷草转氨酶的活性的升高以及高密度脂蛋白胆固醇的含量的降低。
进一步地,基于以上技术方案,所述的食品或药物能够改善酒精成瘾后的戒断反应引起的抑郁、焦虑样行为。
进一步地,基于以上技术方案,所述的食品或药物能够抑制酒精成瘾后的戒断反应引起的促肾上腺皮质激素释放激素、促肾上腺皮质激素和皮质酮的升高。
进一步地,基于以上技术方案,所述的食品或药物能够抑制酒精成瘾后的戒断反应引起的单胺类递质多巴胺和去甲肾上腺素的持续下降。
本发明相对于现有技术具有的有益效果如下:
本发明的组合物由石斛、黄精和葛根配伍组成。本发明的组合物对小鼠酒精性肝损伤有显著的保护作用,包括对肝脏细胞形态、结构的改善,对肝功能以及肝脏脂代谢能力的改善。另一方面,本发明的组合物能改善酒精戒断后的抑郁、焦虑样反应,降低酒精戒断引起的促肾上腺皮质激素释放激素、促肾上腺皮质激素和皮质酮的升高;抑制酒精戒断引起的单胺类递质多巴胺和去甲肾上腺素的减少。
附图说明:
为了更清楚地说明本发明实施例,下面将对实施例涉及的附图进行简单地介绍。
图1为小鼠的酒精偏好率;
图2为组合物对酒精成瘾小鼠肝组织的保护作用;
图3为组合物对酒精戒断小鼠抑郁、焦虑样情绪的改善作用;
图4为组合物对酒精戒断小鼠下丘脑-垂体-肾上腺轴(HPA)轴激素水平的影响;
图5为组合物对酒精戒断小鼠单胺类神经递质的影响;
空白对照组control,戒断组withdrawal,葛根组withdrawal+PE,组合物withdrawal+PDP
**p<0.01,与空白对照组相比有极显著性差异,##p<0.01,与戒断组相比有极显著性差异,&p<0.05,与葛根组相比有显著性差异,&&p<0.01,与葛根组相比有极显著性差异。
具体实施方式
下面结合非限制性实施例对本发明的技术方案做进一步的说明,以使本领域的普通技术人员更全面地理解本发明,但不以任何方式限制本发明。
实施例1组合物的制备
1.1石斛提取物的制备
取100g含水量为85%新鲜石斛冲洗干净后,切分成1cm小段,加入200ml纯净水于榨汁机中破碎10分钟,过滤,收集滤液。向过滤后的滤渣中加入100ml纯净水,少量多次冲洗残留的滤渣,并在60℃条件下,超声20分钟,过滤,合并2次滤液,在3000r/min 条件下,离心15分钟,小心吸取上清液。旋转蒸发仪中浓缩(80℃),最后制成0.5g/ml 石斛提取液,4℃低温保存,备用。
1.2黄精提取物的制备
取60g干燥的黄精药材,清水冲洗干净后,加入180ml纯净水浸泡3h,然后一并转入破壁机内,破碎10分钟。将破碎后汁液及碎渣转移至大烧杯中,90℃的条件下,水浴温煮3h,静置至室温后,过滤,收集滤液。然后向滤液中边搅动边缓慢加入95%乙醇(保证药液中醇浓度为75%-80%,除去蛋白质、无机盐、淀粉等杂质),避免局部乙醇浓度过高造成有效成分包裹损失。充分搅拌后静置,在3000r/min条件下离心15分钟,除去不溶性杂质,小心收集上清液。旋转蒸发仪中浓缩(80℃)浓缩,最后制成0.33g/ml黄精提取液,4℃低温保存备用。
1.3复方组合物的制备
组合物比例为石斛提取液:黄精提取液:葛根粉=10:9:2,组合物质量浓度为0.5g/ml。
实施例2慢性酒精成瘾模型建立
为了更好模拟人类饮酒习惯,采用双瓶自由选择饮酒6周的方式,建立小鼠酒精成瘾模型。在整个实验中,对照组同时提供两瓶饮用水,模型组同时提供水和酒精溶液,并通过设置梯度酒精浓度的方式(酒精浓度3%-10%),促进小鼠饮用更多的酒精溶液。实验中,每天测量饮酒体积和水体积,计算小鼠酒精偏好率。通过测定小鼠前六周及最后一周酒精偏好率分析发现,小鼠饮酒量基本维持稳定,并随着饮酒时间的增加呈现稳定上升的趋势,说明酒精成瘾模型建立成功(如图1所示)。
实施例3对酒精成瘾小鼠肝脏的保护作用
将雄性健康ICR小鼠随机分成对照组(control)、酒精成瘾组(EtOH)、葛根组(EtOH+PE)、复方组合物组(EtOH+PDP),每组12只,按照实施例2的双瓶饮酒法,不同组分别给予生理盐水、酒精、酒精+葛根、酒精+组合物。葛根组灌胃溶有葛根粉的生理盐水,相当于葛根粉5g/kg进行灌胃。组合物组提取物终浓度为0.5g/ml,小鼠灌胃剂量 10ml/kg,相当于原药材5g/kg。
实验结束后,处死小鼠,取小鼠肝脏,将肝进行苏木精-伊红染色(HE)以检查组织学变化。正常对照组小鼠肝细胞的排列呈放射条索状,肝细胞结构完整,胞核结构清楚,边界清晰,没有出现坏死(图2A);酒精组肝细胞排列不整齐,细胞肿胀,胞质出现脂肪空泡化,细胞间隙有油脂脂滴,部分肝细胞核萎缩变形,甚至消失,可见坏死,说明长时间的酒精对肝细胞会造成严重影响(图2B);葛根组改善肝细胞损伤,减少细胞肿胀和萎缩(图2C);同时组合物处理治疗后,脂肪空泡化改善,炎性细胞浸润减少(图2D)。以上实验结果说明酒精对肝脏造成了严重损伤,葛根及复方组合物有效缓解了其损伤,对肝脏具有一定的保护作用。
谷丙转氨酶(ALT)和谷草转氨酶(AST)是机体内重要转氨酶,采用酶联免疫吸附法(ELISA)检测小鼠肝脏的ALT和AST活性,结果如表1所示。相对于空白对照组,酒精(EtOH)处理使小鼠血清中AST和ALT的活性增强。饲喂组合物或葛根后,小鼠血清中AST和ALT的活性明显下降,而且复方组合物(EtOH+PDP)降低ALT、AST水平与单用葛根组(EtOH+PE)相比有极显著性差异,分别降低了23.8%和28.5%。以上说明酒精处理对肝细胞造成损伤,同时反映了与单用葛根相比,组合物对酒精性肝损伤的保护作用更明显。
采用酶联免疫吸附法(ELISA)法检测小鼠的高密度脂蛋白胆固醇(HDL-C)变化情况,结果如表1所示。相对于对照组(control),持续饮酒(EtOH)小鼠的HDL-C含量显著下降,复方组合物(EtOH+PDP)显著提高HDL-C水平,并且与单用葛根(EtOH+PE) 组相比有极显著性差异(升高29.5%)。
表1组合物对酒精成瘾小鼠肝功能的影响(X±SD)
$$p<0.01,vs.control group;
**p<0.01,vs.EtOH group;
&&p<0.01vs.PE group
实施例4组合物对酒精成瘾小鼠戒断后的抑郁、焦虑样反应的改善作用
雄性健康ICR小鼠,分成空白对照组(control,12只)、酒精组(36只)。首先按照实施例2的双瓶饮酒法建立酒精成瘾模型,之后将这些小鼠平均分成3组,实行酒精戒断 7天:戒断组(withdrawal)、葛根组(withdrawal+PE)、组合物组(withdrawal+PDP)。期间,戒断组给予生理盐水,葛根组灌胃葛根粉(溶于矿泉水中),相当于原药材5g/kg,组合物组按10ml/kg给予小鼠灌胃(相当于原药材5g/kg)。第8天实验结束后,进行行为学及其相关生化检测分析。
1、组合物对小鼠酒精成瘾后戒断行为的影响(强制游泳实验FST)
强制游泳实验评估小鼠的抑郁样行为。小鼠被分别放入直径17厘米的烧杯中,水的深度为使尾巴不会接触到烧杯的底部。将各组小鼠依次放入烧杯中心处后开始计时,每次试验共进行5分钟,录像并记录最后4分钟小鼠“绝望行为”的时间,即在水面上漂浮,四肢静止不动,放弃挣扎的总静止时间。实验结果如图3A所示,与对照组相比,酒精戒断(withdrawal)后小鼠不动时间明显延长,出现明显的抑郁样情绪。通过同期给予组合物(withdrawal+PDP),静止时间显著下降,与酒精戒断模型组(withdrawal)相比,有极显著性差异(下降了42.7%)。而单用葛根组(withdrawal+PE)小鼠,与酒精戒断模型组相比,没有显著性差异(p>0.05)。
2、组合物对小鼠酒精成瘾后戒断行为的影响(旷场实验OFT)
旷场实验用于评估小鼠焦虑样行为。实验开始前,准备一个密闭的盒子(40×40×40厘米),平均分为25个格。实验开始后,保持全程安静,先把小鼠放置于测试箱的正中心位置,适应环境1min后,记录小鼠后四分钟在中央区域停留时间与出入频率。有焦虑样倾向的小鼠会减少在旷场中心的时间和频率。实验结果如图3B和图3C所示,与对照组相比,戒断组(withdrawal)小鼠表现明显焦虑样行为;组合物组小鼠(withdrawal+PDP)的焦虑样行为得到极显著改善,在中心区域出现的时间和频率分别增加了了47.9%和55.6%,而单用葛根(withdrawal+PE)时中心区域出现的时间和频率与戒断模型组(withdrawal)相比没有显著性差异(p>0.05)。
实施例5对酒精成瘾后戒断小鼠下丘脑-垂体-肾上腺轴(HPA)的影响
下丘脑-垂体-肾上腺轴(HPA)亢进也是导致酒精中毒的主要机制。当外界酒精信号经神经中枢传达到下丘脑室旁核后,可刺激下丘脑合成促肾上腺皮质激素释放激素(CRH), CRH进而激活垂体前叶分泌促肾上腺皮质激素(ACTH),增多的ACTH又通过刺激肾上腺皮质合成及分泌CORT,导致HPA轴的应激反应,情绪持续亢进,引起相应脑区神经元坏死,最终引发抑郁或焦虑样情绪。行为学实验结束后,***麻醉,处死小鼠,取血,酶联免疫吸附法(ELISA)检测相关激素变化。结果如图4所示,酒精戒断(withdrawal) 后可上调CRH、ACTH和CORT水平,损害HPA轴的反馈作用。而给予组合物 (withdrawal+PDP)后CRH、ACTH和CORT水平显著下降,与模型组(withdrawal)相比,分别下降了44.3%(图4A),28.7%(图4B)和20.9%(图4C),以上结果表明此组合物可能通过调节HPA水平,发挥调节戒断后情绪作用,单独葛根组(withdrawal+PE) 与模型组(withdrawal)相比,对CRH、ACTH和CORT激素水平没有明显的调节作用(p> 0.05)。
实施例6对酒精成瘾后戒断小鼠单胺类神经递质的影响
神经递质是神经元间相互联系的桥梁,参与机体运动、睡眠、情感、学***显著上升了66.6%(图5A),NE水平上升了48.2%(图5B),均比模型组有极显著提高。单独葛根组(withdrawal+PE)与戒断组(withdrawal)相比,对单胺类神经递质没有显著调节作用(p>0.05)。此组合物可能通过上调单胺类神经递质起到一定调节抑郁、焦虑样情绪的作用。
最后应说明的是:以上各实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述各实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解:其依然可以对前述各实施例中所记载的技术方案进行修改,或者对其中部分或者全部技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明各实施例中的技术方案的范围。
Claims (6)
1.一种用于酒精使用障碍的药食同源组合物,其特征在于,由以下原料组成:石斛提取液9-12份,黄精提取液8-10份,葛根粉1-3份;
所述的石斛提取液是通过以下方法制备得到的:取含水量在75-90%的新鲜石斛,加水至新鲜石斛和水的质量比为1:1-1:3,于榨汁机中破碎5-15分钟,过滤,收集滤液;向滤渣中加水至滤渣和水的质量比为1:0.8-1:15,在45-65℃条件下,超声15-25分钟,过滤,收集滤液,合并2次的滤液,静置;然后在3000-5000r/min条件下,离心10-20分钟,收集上清液,浓缩,制成0.4-0.6g/ml石斛提取液,低温保存备用;
所述的黄精提取液是通过以下方法制备得到的:取干燥的黄精药材,加水至黄精和水的质量比为1:2-1:8,浸泡3-5h,然后转入破壁机内,破碎6-12分钟;在70-90℃的条件下水浴温煮1-4h,过滤,向滤液中加入乙醇,使滤液中乙醇终浓度为70-80%,充分搅拌后静置;然后在3000-5000r/min条件下离心10-20分钟,收集上清液,浓缩,制成0.3-0.5g/ml黄精提取液,低温保存备用。
2.权利要求1所述的药食同源组合物在制备改善酒精性肝损伤和/或酒精成瘾后的戒断反应的食品或药物中应用。
3.根据权利要求2所述的应用,其特征在于,所述的食品或药物能够抑制酒精引起的谷丙转氨酶和谷草转氨酶活性的升高以及高密度脂蛋白胆固醇含量的降低。
4.根据权利要求2所述的应用,其特征在于,所述的食品或药物能够改善酒精成瘾后的戒断反应引起的抑郁样、焦虑样行为。
5.根据权利要求2所述的应用,其特征在于,所述的食品或药物能够抑制酒精成瘾后的戒断反应引起的促肾上腺皮质激素释放激素、促肾上腺皮质激素和皮质酮的升高。
6.根据权利要求2所述的应用,其特征在于,所述的食品或药物能够抑制酒精成瘾后的戒断反应引起的单胺类递质多巴胺和去甲肾上腺素的下降。
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