CN106343123B - 一种保健巧克力 - Google Patents
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- CN106343123B CN106343123B CN201610725495.3A CN201610725495A CN106343123B CN 106343123 B CN106343123 B CN 106343123B CN 201610725495 A CN201610725495 A CN 201610725495A CN 106343123 B CN106343123 B CN 106343123B
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Abstract
本发明公开了一种保健巧克力,包括活性成分和基质,所述的活性成分包括以下重量份数的组分:葛根素26‑33份、槲皮素4‑12份、甘草酸1‑7份和原花青素4‑12份和枳椇皂苷26‑33份;所述基质为巧克力,重量份数为50‑65份。其中活性成分葛根素、槲皮素、甘草酸、原花青素和枳椇皂苷在减轻酒精神经毒性以及抑制酒精引起的神经炎症中有明显效果,而巧克力作为基质能够保证口感,易于食用消化。
Description
技术领域
本发明属于保健品领域,具体地说,具体涉及一种保健巧克力。
背景技术
酒精即乙醇是一种亲神经性毒物, 具有脂溶性和水溶性,进入体内的乙醇 10min 后即进入大脑,对人的大脑有直接神经毒性作用,可使血脑屏障通透性增加致使中枢神经严重损害,对中枢神经系统有抑制作用,当酒精进入人神经细胞膜类脂层时,就开始起破坏性作用,经细胞脱水、变性、坏死、缺失,神经细胞体萎缩、树突减少,从而导致大脑萎缩,特别是对青少年的脑损伤更为严重。
现代社会中,随着人们生活节奏的加快,生活压力的增加,人际交往的增多及精神紧张等引起长期饮酒的人数越来越多,大量饮酒,特别是烈性酒,会对人体的消化、神经、循环等系统产生严重的损伤,严重者会导致酒精中毒。
酒精中毒带来的长期和严重的神经炎症所面临的现实现状却是十分令人担忧的,因为绝大多数慢性酒精中毒的患者并没有对其有足够的重视,并得到及时的治疗。在中国的社会现状中,“喝多了”只要睡一觉就行了,其背后隐藏的神经炎症和CNS损伤却给患者的健康带来了长久的影响。
因此,现有技术中亟需一种针对饮酒患者神经炎症的保健产品,能够使得酒精患者脱离宿醉状态,同时抑制酒精导致的神经炎症。
发明内容
为解决上述技术问题,本发明的提供一种以葛根、沙棘、甘草、蓝莓、越橘提取物为活性成分和巧克力为基质的保健品,能够有效预防饮用酒精引发的神经炎症。
本发明的目的是通过以下技术方案的内容来实现的:
一种保健巧克力,包括活性成分和基质,所述的活性成分包括以下重量份数的组分:葛根素26-33份、槲皮素4-12份、甘草酸1-7份、原花青素4-12份和枳椇皂苷26-33份;所述基质为巧克力,重量份数为50-65份。
进一步的技术方案,所述的活性成分和基质的重量份数为:葛根素27份、槲皮素5份、甘草酸3份和原花青素6份和枳椇皂苷27份,巧克力为55份。
进一步的技术方案,所述的巧克力为纯可可脂。
进一步的技术方案,所述的活性成分的纯度为98%。
进一步的技术方案,所述的保健巧克力还包括辅料,所述的辅料根据的不同剂型的需要添加,剂型为丸剂、片剂、颗粒剂、胶囊中的一种。
本发明的保健巧克力在减轻酒精神经毒性以及抑制酒精引起的神经炎症方面的用途 。
本发明作用原理为:酒精慢性处理可以促进神经元预警素的释放,作用于小胶质细胞上的TLR4受体,导致小胶质细胞激活,ROS大量增加,释放多种炎症因子。这在慢性酒精中毒导致神经炎症的过程中发挥关键的作用。本发明中的葛根素、槲皮素、甘草酸、原花青素和枳椇皂苷可以通过阻断上述环节而发挥解酒和抑制神经炎症现象的发生的效果。
有益效果:
本发明一种保健巧克力,其中葛根素、槲皮素、甘草酸、原花青素和枳椇皂苷作为解酒的活性成分能够抑制酒精导致的神经炎症,大大缩短酒精患者的醉酒时间并抑制酒精对脑的损伤,为长期饮酒的人群提供充分的保护作用。
1.本发明中葛根素能够有效在酒精中毒引起的神经炎症中起到好的缓解效果。
2.现有技术中的原花青素广泛使用在化妆品和保健品中,主要有美白抗氧化作用,但还没有研究证明原花青素可以改善酒精中毒引起的神经炎症,而本发明通过研究发现原花青素具备该用途,并且与其他成分协同使用,效果更佳明显。
3.本发明中枳椇皂苷在发挥保肝解酒作用上有非常明显的效果,可以显著改善酒精导致的肝纤维化,降低脂质过氧化,加快乙醇代谢的作用。
4.本发明的甘草酸对HMGB1的激活和释放具有抑制作用,甘草酸可以作为HMGB1的直接或间接抑制剂起到抑制HMGB1,从而抑制胶质细胞活化的作用。 5.本发明的槲皮素可以通过抑制HSP70的释放,降低对小胶质细胞的刺激和激活,从而达到对神经炎症的抑制作用。
6.由于饮酒后经常发生的低血糖症,需要及时补充充足葡萄糖,作为基质的巧克力能够及时给酒精中毒患者补充血糖,缓解醉酒状态。巧克力优选纯可可脂,保证不添加任何添加剂,和其他活性成分组合具有协同作用,更加有利于在胃肠粘膜形成保护层,并且保证产品在胃黏膜均匀散开。
附图说明
图1为实施例1灌胃给药模式下,本发明活性成分对酒精致小鼠翻正反射消失时间的作用示意图。
图2为实施例1灌胃给药模式下,本发明活性成分对酒精致小鼠平衡失调的作用示意图。
图3为实施例1本发明活性成分显著抑制慢性酒精中毒小鼠脑内海马区小胶质细胞的活化。
图4为实施例1本发明活性成分显著抑制酒精刺激诱导的神经元预警素HSP70、HMGB1的释放。
具体实施方式
下面结合实施例和附图对本发明作进一步详细的说明。
实施例1
本发明按照如下重量份数的活性成分进行给药:葛根素27份、槲皮素5份、甘草酸3份和原花青素6份和枳椇皂苷27份,巧克力为55份。配制不同浓度的产品,分为高(80mg/kg)、中(40mg/kg)、低(20mg/kg)三个不同的给药浓度。另外可以根据不同剂型添加不同量的巧克力用以配制成不同的保健巧克力。
上述活性成分纯度均为98%。无水乙醇,购自南京化学试剂有限公司。
本实施例采用直接使用乙醇的研究结果,专业领域技术人员可以使用市售其他酒类重现类似的研究结果。
实验1、上述活性成分配方延长小鼠急性醉酒实验后的清醒时间。
a)试验动物:
健康雄性ICR小鼠,清洁级,体重为18~22 g。实验动物饲养于 12h-12h 昼夜交替的独立环境中,室温维持在24±2℃,自由饮水和摄食,在适应环境 1 周后进行实验。对动物的所有处理均遵循国际疼痛研究协会伦理委员会的要求。
b)试验方法:
小鼠灌胃给药:固定小鼠,使其头、颈和身体呈一直线,使用12号灌胃针头,针头从小鼠的嘴角进入,压住舌头,抵住上颚小心的向内推进,进入食道后会有一种刺空感即可推药。
活性成分配方和乙醇给药剂量和方法:活性成分配方用生理盐水溶解,乙醇浓度为60%,给药方式均为灌胃给药。
小鼠急性醉酒时间测定:将雄性ICR小鼠分为空白对照组、模型组、模型给药组以及正常给药组。模型组(急性醉酒模型):灌胃12ml/kg 60%乙醇溶液;模型加给药组(高中低剂量):禁食(不禁水)12小时后,活性成分配方灌胃给药,20分钟后灌胃12ml/kg 60%乙醇溶液;正常给药组:在基础情况下灌胃给于活性成分配方(80mg/kg)。观察记录小鼠60分钟内的清醒只数随时间的变化情况(清醒即为具有翻正反射)。
c)试验结果描述
如图1所示:空白组(sham)给于0.2ml生理盐水;模型组(alcohol)灌胃12ml/kg60%乙醇溶液;模型加给药组(高中低剂量)灌胃给药,20分钟后灌胃12ml/kg 60%乙醇溶液;单给药组在基础情况下灌胃给于活性成分(高剂量)。记录小鼠60分钟内的清醒只数随时间的变化情况(n=8)。
清醒小鼠只数结果显示,本活性成分配方可以延长饮酒后清醒时间。以上结果表明:本活性成分配方能够显著起到醒酒作用。
实验2、上述活性成分配方改善酒精导致的小鼠平衡失调。
a)试验动物:
健康雄性ICR小鼠,清洁级,体重为18~22 g。实验动物饲养于 12h-12h 昼夜交替的独立环境中,室温维持在24±2℃,自由饮水和摄食,在适应环境 1 周后进行实验。对动物的所有处理均遵循国际疼痛研究协会伦理委员会的要求。
b)试验方法:
小鼠灌胃给药:固定小鼠,使其头、颈和身体呈一直线,使用12号灌胃针头,针头从小鼠的嘴角进入,压住舌头,抵住上颚小心的向内推进,进入食道后会有一种刺空感即可推药。
活性成分配方和乙醇给药剂量和方法:活性成分配方用生理盐水溶解,乙醇浓度为60%,给药方式均为灌胃给药。
平衡失调-转棒实验:将雄性ICR小鼠分为空白对照组、模型组、模型给药组以及正常给药组。模型组:灌胃12ml/kg 60%乙醇溶液;模型加给药组(高中低剂量):禁食(不禁水)12小时后,活性成分配方灌胃给药,20分钟后灌胃12ml/kg 60%乙醇溶液;正常给药组:在基础情况下灌胃给于活性成分配方(80mg/kg)。观察记录给于乙醇溶液10分钟后,在3分钟内未从转棒上坠落的小鼠数。
c)试验结果描述
如图2所示:空白组(sham)给于0.2ml生理盐水;模型组(alcohol)灌胃12ml/kg60%乙醇溶液;模型加给药组(高中低剂量)灌胃给药,20分钟后灌胃12ml/kg 60%乙醇溶液;单给药组在基础情况下灌胃给于活性成分(高剂量)。记录给于乙醇溶液10分钟后,在3分钟内未从转棒上坠落的小鼠数(与空白对照组相比, **P < 0.01,与模型组相比, #P <0.05,##P < 0.01,n=8)。
转棒实验结果显示,本活性成分配方和生理盐水不影响雄性小鼠的平衡感,活性成分配方可以改善醉酒导致的小鼠平衡能力的降低。
实验3、上述活性成分显著抑制小鼠醉酒实验后脑内海马区小胶质细胞的活化。
a)试验动物:
健康雄性ICR小鼠,清洁级,体重为18~22 g。实验动物饲养于 12h-12h 昼夜交替的独立环境中,室温维持在24±2℃,自由饮水和摄食,在适应环境 1 周后进行实验。对动物的所有处理均遵循国际疼痛研究协会伦理委员会的要求。
b)试验方法:
小鼠灌胃给药:固定小鼠,使其头、颈和身体呈一直线,使用12号灌胃针头,针头从小鼠的嘴角进入,压住舌头,抵住上颚小心的向内推进,进入食道后会有一种刺空感即可推药。
活性成分配方和乙醇给药剂量和方法:活性成分配方用生理盐水溶解,剂量为40mg/kg,乙醇浓度为60%,给药方式均为灌胃给药。
将雄性ICR小鼠分为空白对照组、模型组、模型给药组以及正常给药组。模型组:灌胃12ml/kg 60%乙醇溶液;模型加给药组:禁食(不禁水)12小时后,活性成分配方灌胃给药,20分钟后灌胃12ml/kg 60%乙醇溶液;正常给药组:在基础情况下灌胃给于活性成分配方。连给7天,第7天给药后12小时后取小鼠海马区样品。加入含磷酸酶和蛋白酶抑制剂及 PMSF(均为 1:100 加入) 的RIPA 裂解液 300uL。于冰上匀浆,4°C 13000 rpm 离心 15 min 取上清,BCA 法测定样本中的蛋白浓度,加入蛋白上清体积四分之一的 5×loading buffer,煮沸 5 min。
取上述各组蛋白样本,进行免疫印迹试验。每孔上样20μg总蛋白,经聚丙烯酰胺凝胶电泳,转移至PDVF膜上,6% BSA封闭液室温封闭2 h,加入Iba-1一抗(1:1000稀释,CST公司),4 °C摇晃过夜,TBST洗3次,每次10 min,加入相应二抗(1:4000稀释,CST公司),室温孵育2 h,TBST洗3次,每次10 min,显色,采用quantity one软件进行数据分析,实验结果见图3。
c)试验结果描述
如图3所示:免疫印迹试验结果显示,本活性成分配方和生理盐水不会导致小鼠海马区小胶质细胞的过度激活,活性成分配方可以改善醉酒导致的小鼠海马区小胶质细胞激活(图3)。
实验4、本发明显著抑制酒精刺激诱导的神经元预警素HSP70、HMGB1的释放。
a)SH-SY5Y细胞培养
SH-SY5Y细胞采用含10%胎牛血清的F12/MEM培养基培养,加入1%的青霉素和链霉素双抗(购自invitrogen公司)。
b)试验方法:收集生长良好的SH-SY5Y细胞,计数后铺6孔板,每孔细胞数达到1×106,换为无血清培养基,每孔为一组,分为空白对照组、模型组、模型给药组(给药剂量100μM)以及正常给药组(100μM),给药后20分钟后除空白孔外每孔加入无水乙醇,使乙醇终浓度达到0.8%,空白对照组(sham)不给药,给予乙醇后24小时后分别收集上清,使用甲醇-氯仿法提取上清中蛋白,加入2×Loading buffer,沸水浴5min煮样,样本存于-40℃。
取上述各组蛋白样本,进行免疫印迹试验。每孔上样20μg总蛋白,经聚丙烯酰胺凝胶电泳,转移至PDVF膜上,6% BSA封闭液室温封闭2 h,加入HSP70、HMGB1一抗(1:1000稀释,CST公司),4 °C摇晃过夜,TBST洗3次,每次10 min,加入相应二抗(1:4000稀释,CST公司),室温孵育2 h,TBST洗3次,每次10 min,显色,采用quantity one软件进行数据分析,实验结果见图4。
c)试验结果描述:
图4表明,与空白对照组(sham)相比,酒精刺激组HSP70、HMGB1水平显著升高,组方药物给药后可显著降低由酒精刺激引起的HSP70、HMGB1的外排增加(P<0.05),单独加入药物不影响细胞HSP70、HMGB1的外排,提示本组方制剂可以抑制神经元内由酒精刺激引起的HSP70、HMGB1的外排显著增加。
实施例2
一种保健巧克力,包括活性成分和基质,所述的活性成分包括以下重量份数的组分:葛根素26份、槲皮素4份、甘草酸1份、原花青素4份和枳椇皂苷26份;所述基质为纯可可脂,其重量份数为50份。
实施例3
一种保健巧克力,包括活性成分和基质,所述的活性成分包括以下重量份数的组分:葛根素33份、槲皮素12份、甘草酸7份、原花青素12份和枳椇皂苷33份;所述的基质为巧克力,其重量份数为65份。
以上实施例有助于本领域的技术人员进一步理解本发明,但不以任何形式限制本发明。应当指出的是,对本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进都属于本发明的保护范围。
Claims (5)
1.一种保健巧克力,其特征在于,包括活性成分和基质,所述的活性成分包括以下重量份数的组分:葛根素26-33份、槲皮素4-12份、甘草酸1-7份、原花青素4-12份和枳椇皂苷26-33份;所述基质为巧克力,重量份数为50-65份。
2.根据权利要求1所述的一种保健巧克力,其特征在于, 所述的活性成分和基质的重量份数为:葛根素27份、槲皮素5份、甘草酸3份和原花青素6份和枳椇皂苷27份,巧克力为55份。
3.根据权利要求1或2所述的一种保健巧克力,其特征在于,所述的巧克力为纯可可脂。
4.根据权利要求1或2所述的一种保健巧克力,其特征在于,所述的活性成分的纯度为98%。
5.根据权利要求1所述的一种保健巧克力,其特征在于:所述的保健巧克力还包括辅料,所述的辅料根据的不同剂型的需要添加,剂型为丸剂、片剂、颗粒剂、胶囊中的一种。
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