CN113713049A - 用于辅助抗疲劳的中药复方制剂 - Google Patents
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Abstract
用于辅助抗疲劳的中药复方制剂,由黄芪、香橼、天麻、茯苓、白扁豆、百合和甘草组成,其中以重量份计,黄芪:15~30份;香橼:5~15份;天麻:10~20份;茯苓:10~20份;白扁豆:5~15份;百合8~16份;以及甘草:3~10份。本发明的中药复方制剂可以作为辅助抗疲劳的药食两用复方,尤其适用作绿色保健饮料产品。
Description
技术领域
本发明涉及一种可用于辅助抗疲劳的中药复方制剂。
背景技术
疲劳的中医症候分型主要为形体疲劳证、脏腑疲劳证和神志疲劳证三大类型,并有12种常见疲劳证候:筋肉疲劳、关节骨疲劳、神志疲劳、脾胃失调、气虚、血虚、气血两虚、肾元亏虚、阳虚,阴虚、阴阳两虚和月经失常。
疲劳的病因病机复杂,总结我国古籍《素问·调经论》、《脾胃论》、《黄帝内经》、《证治汇补·劳倦》、《素问·调经论》、《何氏虚劳心传》、《景岳全书》、《证治要诀及类方·虚损门》、《素问·示从容论》等可知起居失常、偏食或暴饮暴食、情志不舒、过度疲劳、个人体质等皆是疲劳诱因,疲劳的病机在于机体气血之损与脾肾虚弱、肝气郁滞等脏腑功能下降或失调有关。另外《素问·调经论》、《何氏虚劳心传》等亦指出过劳伤阴、脾失健运、湿热内生,湿热郁结于肝肾等部位,导致津液耗伤、成痰、成瘀、伤肝脾肾等,以至形成“阴虚内热”之表证。
发明内容
本发明的目的是提供一种可药食两用的辅助抗疲劳的中药复方制剂。
根据本发明的第一方面,提供了一种用于辅助抗疲劳的中药复方制剂,由黄芪、香橼、天麻、茯苓、白扁豆、百合和甘草组成,其中以重量份计,黄芪:15~30份;香橼:5~15份;天麻:10~20份;茯苓:10~20份;白扁豆:5~15份;百合8~16份;以及甘草:3~10份。
本发明的复方制剂可以采用多种剂型,例如水提原液形式或添加辅料并干燥后制成的颗粒冲剂或胶囊等形式。辅料可以采用可溶性淀粉和糊精混合物等。
根据本发明的另一方面,提供了一种可用于辅助抗疲劳的保健饮品,包含上述中药复方制剂。
发明人通过多年精心筛选,形成了一种安全有效的用于辅助抗疲劳的中药复方:首先采用调养多个脏腑的同时结合补益气血的药味;另外在补益的同时,加入调理气机、疏肝解郁、清热滋阴的药味,达到调理肝肾阴虚、健脾利湿、理气扶正、标本兼治的效果,从而能够有效缓解体力疲劳。
附图说明
图1为不同组的体重记录曲线;
图2为不同组的肝肾、肌肉指数柱形图;
图3为不同组的小鼠负重力竭游泳时间的柱形图;
图4A-C分别为不同组的小鼠肝/肌糖原及血糖含量柱形图;
图5A和B分别为不同组游泳实验后小鼠血尿素氮含量及血乳酸曲线下面积柱形图;
图6A-D分别为不同组游泳实验后小鼠血清乳酸脱氢酶、肌酸激酶、谷丙转氨酶、谷草转氨酶含量柱形图;
图7A-E分别为不同组游泳实验后小鼠肝脏超氧化物歧化酶SOD、谷胱甘肽过氧化物酶GSH-Px活性及MDA含量柱形图,其中A:肝SOD活性,B:肝GSH-Px,C:肝MDA,D:肌SOD,E:肌MDA。
所有图中统计学显著性分析表示为:a,与正常空白组比较,p<0.05;b,与羧甲基茯苓多糖组比较,p<0.05。
具体实施方式
下面结合具体实例和附图对本发明做进一步说明,本领域技术人员应该理解,实例和附图只是为了更好地理解本发明,并不用来做出任何限制。
制备中药复方原液
按重量份称取黄芪60g、香橼30g、天麻45g、茯苓45g、白扁豆30g、百合36g、甘草18g(均为市购),加入总药味8倍量的饮用水,即2112mL水,常温浸泡0.5h,每次煎煮沸腾1h,将提取液室温放凉后,采用8层纱布进行过滤,收集滤液备用,滤渣再重复提取1次,共提取2次,弃掉滤渣,合并2次的提取液,减压浓缩后获得1056mL原液。
制备复方颗粒制剂
向上述原液中加入32g混合辅料:可溶性淀粉和糊精混合物(二者质量比为4:1),搅拌均匀后喷雾干燥,制得可例如用于冲服的颗粒制剂或固体饮料粉末58g。
复方颗粒制剂指标检测
该复方颗粒制剂的溶解度、粒度、分散性、冲调稳定性、堆密度、休止角、水分和吸湿性等各指标检测结果如表1所示。从表中可以看到:溶解度约为309s,堆密度值约为0.34;休止角约为25.45;水分约为1.12%和吸湿性约为20.43%,粒度要求均能通过六号筛;分散性要求均能通过筛网;冲调稳定性要求离心后无沉淀。制剂的各项理化性质稳定。
表1缓解体力疲劳复方制剂理化指标测定结果
作为保健食品服用时,上述制备的固体饮料粉末的成人推荐剂量可以为:每日一次,每次15g。
动物建模抗疲劳对照实验
实验材料
肌酸激酶测定试剂盒(货号:100020040)、葡萄糖测定试剂盒(货号:100020100)、尿素测定试剂盒(产品批号:190971)、乳酸脱氢酶测定试剂盒(货号:100020050)、天冬氨酸氨基转移酶测定试剂盒(货号:100020013)、丙氨酸氨基转移酶测定试剂盒(货号:100020003)均购自于中生北控生物科技股份有限公司;小鼠全乳酸试剂盒(货号:A019-1-1南京建成生物工程研究所)、肝/肌糖原测试盒(货号:A043-1-1)、谷胱甘肽过氧化物酶测试盒(货号:A005)、丙二醛测试盒(货号:A003-1)、超氧化物歧化酶测试盒(货号:A001-1)均购自于南京建成生物工程研究所有限公司。
阳性药为复方高山红景天口服液(国药准字B20020080,吉林强身药业有限责任公司)。
中药复方制剂则采用上述配方制备的复方原液。
实验动物与分组
SPF级BABL/C小鼠50只,雄性,体质量18~22g,购自北京维通利华实验动物技术有限公司,许可证编号:SCXK(京)2016-0006。本研究由北京协和医学院药用植物研究所实验动物中心伦理委员会批准通过,伦理审查编号:SLXD-20200902022。在北京协和医学院药用植物研究所动物房饲养,每天自由饮水、进食。
所有BABL/C雄性小鼠于常规条件下适应性饲养1周后随机分为5组,分别为空白组(Control)、阳性药复方高山红景天组(Rhc)、缓解体力疲劳复方高剂量组(CH)、缓解体力疲劳复方中剂量组(CM)、缓解体力疲劳复方低剂量组(CL),每组12只。按照人与小鼠用量换算关系计算得到高剂量组给药浓度为0.35g·mL-1,高、中、低剂量分别为人临床用量的10倍、5倍、2.5倍,即该复方中剂量组的浓度为0.18g·mL-1,低剂量组的浓度为0.09g·mL-1。各组均灌胃38d,每天1次。动物每周称重1次,按体重调整受试样品剂量。各组动物处理方法见表2。
表2分组给药方案
实验方法
检测小鼠负重力竭游泳时间、BUN、MG、LG、Glu等与能量代谢有关的指标,并三次断点采血检测了BLA的代谢曲线;检测了肝肌组织SOD、MDA、GSH-Px的含量检测,评价受试物能否减轻氧化应激损伤,从而缓解机体疲劳;检测了血清CK、LDH、AST、ALT含量检测,检验受试物能否减少代谢产物累积,从而达到缓解机体疲劳的功效。
其中,耐力是反映直接反映机体抗疲劳程度的金指标,耐力越强(负重力竭游泳时间越长),抗疲劳能力越强。剧烈运动时,机体相对缺氧,糖酵解加快,产生大量乳酸,使肌肉中H+浓度上升,pH值下降,导致疲劳,减少乳酸的产生或加快乳酸的消除,都可延缓疲劳发生和/或加速疲劳的消除,血乳酸水平可以反映有氧代谢能力、疲劳的产生和消除速度。肝糖原及肌糖原是机体重要的能源储备物质,若复方制剂能显著提高小鼠机体糖原储备,则说明能在一定程度上缓解疲劳。机体在长时间运动时,体内能源储备糖原物质消耗殆尽,就会开始消耗体内蛋白质,从而产生大量BUN。机体对运动负荷的适应性越低,形成的尿素越多。反之,机体运动后BUN含量越低,表明机体对运动负荷的适应能力越强,抗疲劳能力也就越强,其余指标的检测含义在结果分析部分中有详细阐释。
指标检测方法如下:
1.负重力竭游泳时间的测定
连续灌胃第29天时,给药30min后,将尾部负重5%体重铅皮的小鼠置于游泳箱中游泳,水深不少于30cm,水温25±1℃,记录小鼠自游泳开始至力竭而沉入水中并维持8s不能浮出水面,且四肢不能协调划水的时间。
2.全血乳酸含量的测定
连续给药第33天,给药后30min,进行3次断点采血:①静息时,对小鼠进行眼眶取血30μL;②然后不负重,在温度为30℃的水中游泳10min停止,立即采血30μL,③休息20min后再次采血30μL。按全血乳酸测试盒说明书操作,测定吸光度:测定前先做预实验,制备上清液:取待测样本(全血)0.05mL,加入蛋白沉淀剂0.3mL混匀,4000r/min离心20min,取上清。设定空白管、标准管和测定管,分别加入0.01mL的双蒸水、3mmoL/L标准液、上清液,在所有管中分别加入酶工作液0.5mL、显色剂0.1mL,混匀各管后,37℃水浴准确反应10min,最后加入终止液1mL,在530nm波长处测定各孔的OD值。绘制标准回归曲线,计算各样本BLA浓度值。计算三个点的血乳酸含量及血乳酸曲线下面积。
血乳酸曲线下面积=5×(游泳前血乳酸值+3×游泳后0min的血乳酸值+2×游泳后休息20min的血乳酸值)
3.BUN、CK、LDH、Glu、AST、ALT含量的测定
连续灌胃第38天时,即末次给药30min后,让各组小鼠无负重在温度为28±1℃的水中游泳60min,休息30min后,迅速摘眼球取血,4000rpm·min-1离心15min取血清,置于4℃冰箱保存。按照对应试剂盒说明,在全自动生化检测仪中测定BUN、CK、LDH、Glu、AST、ALT的含量。
4.肝/肌组织生化指标含量的测定
摘眼球取血后,迅速脱颈椎处死小鼠,取肝脏和双后肢肌肉,置于冰培养皿上,经生理盐水漂洗后用滤纸吸干,制备肝、肌肉10%组织匀浆,置于-20℃冰箱保存。测定肝脏组织中LG、SOD、MDA、GSH-Px的含量及肌肉组织中MG、SOD、MDA的含量。
实验结果分析
(1)小鼠给药期间,每周称重1次,观察小鼠体重变化,如图1所示,给药38天期间,各组小鼠体重均呈现上升趋势,但灌胃第五周时,CL组较前一周体重略微减少,考虑原因为:给药第29天时小鼠进行负重力竭游泳造成的影响。在给药过程中,给药组小鼠体重与空白组相比,均无显著性差异,各给药组小鼠之间体重亦无显著性差异(P>0.05),未见其他体重异常。小鼠毛发、尿液、粪便正常,社会学行为等正常表明小鼠生长发育良好。
(2)如图2所示各组小鼠肝脏指数、肾脏指数、肌肉指数与空白组相比,均无显著性差异,各给药组小鼠之间亦无显著性差异(P>0.05)
(3)空白组平均力竭时间为646.2s,高、中、低剂量组的平均力竭时间分别为859.6s、826.6s、780s,阳性药组为880.43s。各给药组均能不同程度的延长小鼠运动时间,与空白组相比三个复方剂量组对小鼠负重力竭时间的延长率分别为42.66%、37.19%、29.45%,复方高山红景天口服液可延长小鼠的运动力竭时间46.11%。其中CH、CM组可显著延长小鼠负重力竭游泳时间(P<0.05),增强小鼠的运动耐力,由图3可知,Rhc、CH、CM三组间组无显著性差异。缓解体力疲劳复方对力竭时间呈现剂量依懒性影响,但不显著(P>0.05)。
(4)剧烈运动后肝糖原耗尽,机体血糖水平迅速下降,低血糖抑制中枢神经系统功能,会产生乏力、头晕等疲劳现象,因此降低运动耐力。各个复方剂量给药组均能不同程度的提高机体糖原储备量。空白组LG的平均含量为4.478mg/g,缓解体力疲劳复方三个剂量组LG的平均含量分别为:8.066mg/g、4.898mg/g、8.953mg/g,阳性药组的平均含量为6.885mg/g。如图4A、4C所示与空白组相比,CH、CL组可显著增加肝糖原水平(P<0.05),增加率分别为80.10%、99.90%,三个剂量组增加肝糖原含量的程度由大至小为CL>CH>CM,这一检测结果与血糖检测结果一致:分别增加Glu含量179.20%、96.71%、73.62%。由图4B可知,空白组MG的平均含量为0.63mg/g,缓解体力疲劳复方三个剂量组MG的平均含量分别为:1.103mg/g、0.803mg/g、0.788mg/g,阳性药组的平均含量为1.131mg/g。Rhc、CH组可显著增加机体MG水平(P<0.05),增加率分别为79.57%、75.13%。
所以CH、CL及Rhc通过提高糖原储备量,维持运动后的血糖平衡,维持中枢神经系统功能,缓解体力疲劳的产生,此外受试复方组未表现出剂量依赖性地增加LG/MG及Glu含量。
(5)如图5A所示,空白组BUN的平均含量为14.52mmol/L,缓解体力疲劳复方高、中、低剂量组MG的平均含量分别为12.44mmol/L、11.36mmol/L、13.56mmol/L。复方高山红景天口服液组的BUN含量为11.41mmol/L。各给药组均能不同程度的降低小鼠运动后BUN含量,与空白组小鼠相比,CH、CM组及Rhc组可显著降低BUN含量(P<0.05),从而增加机体对运动负荷的适应能力,缓解体力疲劳。
乳酸在体内的积累取决于乳酸的产生与消除速度,因此,减少乳酸的产生或加快乳酸的消除,都可延缓疲劳发生和/或加速疲劳的消除,血乳酸水平可以反映有氧代谢能力、疲劳的产生和消除速度。空白组BLA曲线下面积为964.77,缓解体力疲劳复方高、中、低剂量组BLA曲线下面积分别为874.25、891.69、913.04,阳性药组为928.53。如图5B所示,与空白组相比,各给药组均可显著降低BLA曲线下面积(P<0.05),CH、CM、CL及Rhc的降低率分别为8.85%、8.263%、5.36%、3.76%。
(6)运动介导的氧化应激使体内活性氧簇(ROS)大量激增,而ROS引起细胞损伤,内容物便释放进入血液,引起机体离子及代谢功能紊乱,引发疲劳。而LDH、CK等大分子可通过损伤的细胞进入淋巴液,最终进入血液循环。LDH不仅是组织损伤的标志,亦是参与糖酵解和糖异生过程中催化乳酸和丙酮酸之间氧化还原反应的重要酶类,直接影响机体能量代谢过程,其活力与机体的乳酸含量存在正相关,当机体的乳酸含量上升时,LDH活力也随之增强,催化运动所产生的乳酸转变为丙酮酸,减少其在肌肉内的堆积,LDH活性增强可以大大加速BLA的清除率,进而提高机体的抗疲劳能力。如图6A所示,LDH活性低在一定程度上反映了乳酸含量低,CH、CL可显著降低LDH活性(P<0.05),其降低率分别为25.92%、17.20%。
同理,血清CK水平的高低与横纹肌受损程度有关,正常生理状态下CK处于肌肉中,使肌酸上增加磷酸基团,转化为高分子磷酸基团,释放能量。肌细胞受损后,CK会经细胞间隙渗透到淋巴液,最终进入血液循环,间接引起血清中CK活力增加,在血液中检测到CK含量增加,暗示肌肉损伤的发生。图6B显示各给药组均可显著降低血清CK活力(P<0.05),改善急性运动引起的骨骼肌损伤,从而缓解体力疲劳。剧烈运动还会导致谷丙转氨酶(ALT)及谷草转氨酶(AST)生理性增高,ALT、AST是肝组织损伤的关键标志物,若复方能显著降低小鼠长时间运动后机体的ALT、AST含量,则说明复方能在一定程度上减轻肝组织损伤程度,缓解小鼠疲劳。如图6C、D与空白组小鼠相比,CH、CM组可显著降低ALT含量,CH可显著降低AST含量(P<0.05)。Rhc可显著降低LDH活性、ALT及AST含量。
(7)剧烈运动容易产生大量自由基,引发机体内某些器官细胞出现脂质过氧化损伤改变生物膜的通透性,破坏细胞结构和功能,并扰乱细胞内外的离子转运,影响肌纤维的兴奋收缩和偶联,也会导致机体产生疲劳。MDA是一种在机体代谢过程中产生的脂质过氧化产物,与机体组织受到活性氧积累而诱发的膜脂质过氧化作用有着密切关联。正常状态下,自由基可被机体自身的抗氧化酶(如SOD和GSH-Px)清除,维持机体氧化/抗氧化平衡,一旦机体抗氧化能力下降,自由基不能被及时清除,平衡被打破很容易加剧疲劳的产生。
处于运动疲劳状态时,机体过氧化产物会增加,并伴随抗氧化酶活性的微小降低趋势。因此,可以通过测定运动后肝脏/肌肉SOD、GSH-Px活力和MDA的含量,评估其缓解体力疲劳作用。如图7A-E所示,与空白组相比,各剂量给药组均有助于提高运动后小鼠机体内SOD、GSH-Px活力并减少脂质过氧化产物MDA的产生,其中CH、CM可显著提高肝脏和肌肉中SOD活力,并降低MDA含量(P<0.05)。CH增强了肝脏和骨骼肌中SOD14.42%和25.51%的活性,CL组可显著增强肝脏中GSH-Px活力(P<0.05),各给药组均可减少肝脏中MDA的产生但效果不显著(P>0.05),CH、CM可显著减少肌肉中MDA含量(P<0.05)。
Rhc组增强了肝脏和骨骼肌中SOD21%以上的活性,并可显著增强肝脏中GSH-Px活力(P<0.05)。高、中剂量复方组及阳性药组通过增强清除机体自由基的能力和减轻脂质过氧化对细胞的损伤,减轻氧化应激损伤,从而起到缓解体力疲劳的作用。
综上所述,复方高、中剂量组可显著缓解体力疲劳,提高小鼠的运动耐力。其作用机制是通过减少疲劳小鼠机体MG/LG消耗量,维持运动后机体Glu的稳定,降低BUN含量,通过增强LDH活性加快运动后乳酸的清除速率,减少BLA积累,即提高机体能量代谢;通过增强SOD/GSH-Px活力,降低脂质过氧化产物MDA的含量来增强清机体除自由基的能力和减轻高强度运动后脂质过氧化对机体的损伤,即增强机体抗氧化能力;通过抑制运动后血清中CK、AST、ALT活力的增加来减轻细胞器、细胞膜的损伤,减少运动代谢产物的积累或加快其清除速率从而缓解疲劳的产生。综合分析CH组为缓解体力疲劳效果的较优剂量,可有效调节机体能量代谢,增强供能,减少运动损伤及代谢产物累积,有效缓解体力疲劳。
因此,本发明的中药复方可以作为抗疲劳药食两用复方,尤其适用作绿色保健饮料产品。
Claims (4)
1.一种用于辅助抗疲劳的中药复方制剂,由黄芪、香橼、天麻、茯苓、白扁豆、百合和甘草组成,其中以重量份计,黄芪:15~30份;香橼:5~15份;天麻:10~20份;茯苓:10~20份;白扁豆:5~15份;百合8~16份;以及甘草:3~10份。
2.根据权利要求1的复方制剂,其中该制剂为水提液形式。
3.根据权利要求1的复方制剂,其中该制剂为颗粒冲剂或胶囊形式。
4.一种用于辅助抗疲劳的保健饮品,包含权利要求1的中药复方制剂。
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