CN112869154A - 一种用于化疗减毒增效的鱼腥草挥发油护心胶囊及其制备方法 - Google Patents
一种用于化疗减毒增效的鱼腥草挥发油护心胶囊及其制备方法 Download PDFInfo
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Abstract
发明涉及一种用于化疗减毒增效的鱼腥草挥发油护心胶囊及其制备方法。应用鱼腥草为原料,经水蒸气蒸馏法提取得到鱼腥草挥发油,按重量计鱼腥草挥发油约20~40份,辅料:山楂提取物10~20份、灵芝提取物5~10份和大豆油40~60份。产品活性效价高,具有心肌保护作用,适合化疗引起的心肌毒性的肿瘤患者使用。鱼腥草挥发油来源于天然食材,安全性好、化学成分明确,具有很好的心肌保护效果;产品辅料中山楂能消食健胃、增进食欲;灵芝扶正固本,适合心肌功能不全人群,尤其是接受化疗引起心脏毒性的肿瘤患者。本品可以作为化疗伴侣,适用于化疗前后,对心肌具有很好的防护作用,能提高肿瘤患者的预后,具有较好的市场前景。
Description
技术领域
本发明属于保健食品技术领域,涉及食品加工技术领域,特别涉及发明一种针对化疗引起心肌毒性,具有减毒增效的鱼腥草挥发油护心胶囊及其制备方法。
背景技术
迈入21世纪,由于食品供给的富余,疾病模式已发生转变,肿瘤已经成为危害全球健康的重要杀手。随着治疗技术的发展和研究领域的深入,肿瘤治疗已经取得飞速发展。现今的肿瘤治疗将外科手术、化疗和放疗等多种方式联合应用,显著提高了肿瘤患者的5 年存活率。但随着大量化疗药物的使用,药物的不良反应,尤其是心血管方面的不良反应,如心功能不全、心肌缺血、高血压、血管栓塞和心律失常等,引起人们越来越多的关注。蒽环类抗生素(如多柔比星)就是这些药物的典型代表,作为一种高效的广谱抗肿瘤抗生素,疗效确定,但是在临床化疗中的严重的心脏毒性反应并呈现出明显的剂量-效应线性关系,大大限制了它的使用。
鱼腥草是一种药食同源的传统食材,在食品、药品等方面有很大的开发前景。鱼腥草挥发油具有抗氧化、抗炎、抗肿瘤、心肌保护和抗病毒等多种药理作用,其药理作用主要与其挥发油中甲基正壬酮、乙酸龙脑酯、棕榈酸、正癸酸等主要成分密切相关。此外,与同类产品相比,鱼腥草挥发油是经水蒸气蒸馏法制得,属于纯天然食材鱼腥草的安全无害提取物,且其化学成分明确,易于被消费者接受,在保健食品方面具有很好的开发空间。
本品成分有灵芝和山楂。灵芝是我国重要的药食兼用资源,具有较好的营养、保健、医疗价值。灵芝的应用范围非常的广泛,其主要原因是灵芝具有扶正固本之功效。现代药理学也证明灵芝具有可保护心脏、抗肿瘤和提高免疫力等功效。山楂也是中国特有的药果兼用资源,具有强心、抗心律不齐、抗癌等作用,同时也是健脾开胃、消食化滞、活血化痰的食材。
发明内容
本发明提供了发明一种用于化疗减毒增效的鱼腥草挥发油护心胶囊及其制备方法,应用药食两用的天然食材,结合现代仪器技术和食品加工技术,提取鱼腥草挥发油,明确该挥发油的化学成分,制备能发挥心肌保护作用,且抑制化疗引起心肌毒性的鱼腥草挥发油护心胶囊。该产品可作为化疗药物的伴侣,可以在化疗前后长期食用,有助于预防和修复心肌损伤,可提高患者的生活质量,对预后具有积极作用。
为了实现以上目的,本发明所提供的技术方案是:
一种用于化疗减毒增效的鱼腥草挥发油护心胶囊,所述护心胶囊包括软胶囊内容物和囊壳,所述软胶囊内容物质量份数的物质组成:按重量计鱼腥草挥发油20~40份,辅料:山楂提取物10~20份、灵芝提取物5~10份和大豆油40~60份。
所述囊壳在胶液配置上选择了明胶100份、甘油35-50份、水80-100份、山梨醇3-7份、棕氧化铁0.5-1份:(1)明胶无嗅无味,不溶于水,但浸泡在水中时,可吸收5~10倍的水而膨胀软化,如果加热,则溶解成胶体,冷却至35~40℃以下,成为凝胶状;(2)甘油食用对人体无毒,是食品加工业中通常使用的甜味剂,具有高活性、抗氧化、促醇化等功效。(3)山梨醇略有甜味,具有吸湿性,能溶解多种金属,能与多种金属形成络合物。(4)棕氧化铁是辅料,着色剂。
一种针对化疗引起心肌毒性的鱼腥草挥发油护心胶囊的制备方法:
选用鱼腥草,提取出活性成分鱼腥草挥发油,与山楂提取物、灵芝提取物和大豆油混匀,制得一种用于化疗减毒增效的鱼腥草挥发油护心胶囊。具体步骤为:
(1)称取干燥燥鱼腥草粉末,装入圆底烧瓶中,加10~40倍蒸馏水,混匀并浸泡10 h后,连接挥发油测定器与回流冷凝管。将整套装置放置于SKM型数显恒温电热套中,温度设置220 ℃,达到微沸状态,并保持微沸10 h,加热回流10 h后停止加热,收集挥发油。
(3)将山楂粉和灵芝粉加10~40倍蒸馏水经水煮2~4小时,煮两次,过滤合并两次滤液,旋转蒸发仪浓缩,减压真空干燥箱干燥,用超微粉碎机进行粉碎,过240-300目筛子,与鱼腥草挥发油及大豆油混匀得软胶囊内容物;
(4)胶液配制:按重量称取明胶100份、甘油35-50份、水80-100份、山梨醇3-7份、棕氧化铁0.5-1份,投入化胶罐中,搅拌融化明胶,控制真空度,减压抽真空,放入保温桶内,保温50-60℃,放置6-8小时,脱气,备用;
(5)制备鱼腥草挥发油软胶囊:将制备好的胶液和软胶囊内容物送入压丸机中,压制出软胶囊;
(6)定型:控制环境温度18-26℃,相对湿度20-35%,转笼定型1-3小时;
(7)洗丸:将得到的胶囊用95%以上的酒精洗去胶囊表面的油渍;
(8)干燥:出丸至脱盘上,送入干燥室,控制环境温度18-26℃,相对湿度20-35%,静态干燥24-36小时后,将胶囊出丸,拣丸(去除非正型丸)。
(9)包装成品:将正常型软胶囊装瓶、封装,即是用于抗化疗引起心肌毒性的鱼腥草挥发油护心胶囊成品。
本发明针对化疗引起心肌毒性的减毒增效的鱼腥草挥发油护心胶囊的市场需求,为丰富鱼腥草在保健的开发应用,提供一种用于化疗减毒增效的鱼腥草挥发油护心胶囊制备方法,以鱼腥草挥发油为主要成分,配以山楂和灵芝极大程度上保护病人的心肌的同时,改善食欲、健胃消食、提高免疫力和扶正固本的性质,制成心肌保护软胶囊产品,有助于化疗引起的心肌损伤预防和修复,具有心肌保护作用,为解决化疗引起心肌毒性等毒副作用提供新产品,促进药食同源天然食材的高值化应用。
附图说明
图1大鼠心脏组织切片病理学观察(HE,×40),其中A为正常组,B为化疗组,C为化疗+挥发油组,D为阳性对照组。
具体实施方式
下面结合具体实施方式对本发明进行进一步的阐述。
实施例1:鱼腥草挥发油的心肌保护作用
1、实验材料
1.1材料与试剂
鱼腥草挥发油通过以下方法获得:参照2010年版中国药典一部附录XD挥发油测定法(甲法)提取挥发油:鱼腥草经粉碎机粉碎成粗粉,室温下浸泡蒸馏水中10 h,料液比为1:10,加热回流10 h,得到的鱼腥草挥发油。GC-MS化学成分鉴定挥发油的主要成分有癸酸(35.63%)、棕榈酸(22.64%)、甲基正壬酮(10.42%)和乙酸龙脑酯(9.95%)等28种化学成分。
多柔比星(DOX:蒽环类常用化疗药物,用于建立心肌毒性动物模型),深圳万乐药业有限公司;乙醇,分析纯,西陇化工股份有限公司;维生素E(VE)、 美国Sigma公司;BCA蛋白浓度测定试剂盒,上海碧云天公司;乳酸脱氢酶(LDH)试剂盒、和过氧化氢酶(CAT)试剂盒,南京建成生物工程研究所有限公司;
实验动物:SD大鼠(180±20 g) 40只,雄性,湖南斯莱克景达实验动物有限公司,使用许可证:SCXK(湘)2016-0002。
1.2主要仪器设备
3K15-冷冻离心机 美国Sigma公司;SHP-150生化培养箱 上海森信实验仪器有限公司;AL-104电子天平 梅特勒-托利多仪器上海有限公司;Thermo 3001 Varioskan Flash全波长多功能酶标仪 美国Thermo公司;Milli-Q超纯水仪 美国Millipore公司;SKM型数显恒温电热套 山东鄄城华鲁电热仪器有限公司。
2 实验方法
2.1动物实验设计与分组
实验分组(每组10只):生理盐水组(正常组),DOX组(化疗组),DOX+挥发油100 mg/kg 组(化疗+鱼腥草挥发油组),DOX+维生素E 500 mg/kg 组(为阳性对照组)。挥发油及VE处理:精密量取挥发油,以吐温-80作乳化剂,将其溶于生理盐水中,配成含0.5 %吐温-80的鱼腥草挥发油母液,临用前将母液稀释至适宜的浓度。实验大鼠适应一周后,每天称重,第一天,正常组和化疗组灌胃生理盐水,其他组灌胃相对应的鱼腥草挥发油或维生素E ;第二天,化疗组,化疗+鱼腥草挥发油组及阳性对照组腹腔注射DOX 20 mg/kg,正常组注射同剂量的生理盐水;第三天和第四天灌胃方式与第一天相同。第五天眼球取血,处死大鼠,取肠内容物和心脏组织。
2.2 心脏组织HE染色 实验结束收集心脏,生理盐水洗净,滤纸吸取表面残液,纵切取一半心脏组织用10%中性福尔马林缓冲液固定。制作心脏组织HE染色,切片脱蜡、切片复水、苏木素染色、伊红染色、脱水封片,再用倒置显微镜进行拍照。
2.3 血清中LDH活力的测定 对大鼠进行摘眼球取血,室温静置2小时,室温3000r/min离心10 min,收集血清,使用LDH试剂盒测定LDH活力(U/L)。
2.4 心脏组织CAT测定 分离大鼠心脏,制备10%心肌组织匀浆,严格按照CAT和试剂盒说明书检测CAT活力,并使用BCA蛋白定量测定法检测蛋白质质量浓度。
2.5 数据处理 实验组数据用平均值±标准偏差(M±SD)的形式表示,采用SPSS23.0软件进行统计学分析。p<0.05时,表示与比较组存在显著差异,p<0.01表示与比较组差异极显著。
3 结果与分析
3.1 挥发油对化疗致心肌损伤大鼠心脏微观结构的影响
心脏组织的组织病理学检查是为了进一步镜下显微观察化疗DOX诱导的心脏毒性。本实验利用HE染色的方式,利用倒置显微镜观察心脏组织切片病理结构,观察视野如图1所示,正常组(A)显示了正常心肌束的结构,心脏结构整齐,心肌纤维排列整齐,结构清晰且正常。化疗组(B)心肌纤维明显断裂,心肌细胞有肿胀反应,结构破坏较明显;而与化疗组(B)相比,化疗+鱼腥草挥发油组(C)和阳性对照组(D)心肌纤维排列明显整齐,纤维断裂减少,心脏结构趋于完整。综上所述,鱼腥草挥发油能有效促进化疗诱导的心肌损伤模型大鼠中心脏组织形态趋于正常化,具有心肌保护作用。
3.2 鱼腥草挥发油对化疗引起心肌损伤的影响
乳酸脱氢酶(LDH)是一种常见的心肌酶,它能催化乳酸和丙酮酸之间的相互转化,其作为心肌诊断的标记物广泛应用于心肌损伤研究中。从表1可以看出,经化疗处理过的大鼠,其血清中LDH活力显著高于正常对照组(p<0.05),说明化疗对大鼠心肌组织损伤严重;而与化疗组相比,阳性对照组LDH活力降低(p<0.05),同时,给予鱼腥草挥发油各组LDH的活力显著降低,表明鱼腥草挥发油具有心肌保护作用可抑制化疗介导的心脏毒性。
表1 大鼠体内LDH和CAT的含量
注:与正常组相比:#p <0.05,与模型组相比:* p <0.05,** p <0.01
3.3 心脏组织中CAT的测定
CAT作为机体中重要的抗氧化物酶,能将自由基H2O2分解成H2O和O2,抑制氧化应激损伤,如表1所示,与正常组相比,化疗组中大鼠心脏组织中CAT活性降低;与化疗组相比,化疗+鱼腥草挥发油组中大鼠心脏组织CAT活性上升(p <0.05)。同时,阳性对照组中CAT活性显著增加(p <0.05)。这些结果提示鱼腥草挥发油的心肌保护作用与其提高大鼠抗氧化防御能力有关。
实施例2:用于化疗减毒增效的鱼腥草挥发油护心胶囊的制备方法
一种用于化疗减毒增效的鱼腥草挥发油护心胶囊的保健食品,其组成包括两部分:软胶囊内容物和囊壳部分。软胶囊内容物按重量鱼腥草挥发油30份、山楂提取物15份、灵芝提取物8份和大豆油50份;囊壳由明胶100份、甘油40份、水90份、山梨醇5份、棕氧化铁0.5份。制备方法的具体操作步骤为:
(1)鱼腥草干品、山楂、灵芝经粉碎机粉碎,过8-10目筛,得鱼腥草粉、山楂粉、灵芝粉;
(2)称取干燥燥鱼腥草粉末,装入圆底烧瓶中,加25倍蒸馏水,混匀并浸泡10 h后,连接挥发油测定器与回流冷凝管。将整套装置放置于SKM型数显恒温电热套中,温度设置220 ℃,达到微沸状态,并保持微沸10 h,加热回流10 h后停止加热,收集挥发油;
(3)将山楂粉和灵芝粉加25倍蒸馏水经水煮3小时,煮两次,过滤合并两次滤液,旋转蒸发仪浓缩,减压真空干燥箱干燥,用超微粉碎机进行粉碎,过300目筛子,与鱼腥草挥发油及大豆油混匀得软胶囊内容物;
(4)胶液配制:按重量称取明胶100份、甘油40份、水90份、山梨醇5份、棕氧化铁0.5份,投入化胶罐中,搅拌融化明胶,控制真空度,减压抽真空,放入保温桶内,保温60℃,放置7小时,脱气,备用;
(5)制备鱼腥草挥发油软胶囊:将制备好的胶液和软胶囊内容物送入压丸机中,压制出软胶囊;
(6)定型:控制环境温度18-26℃,相对湿度20-35%,转笼定型1-3小时;
(7)洗丸:将得到的胶囊用95%以上的酒精洗去胶囊表面的油渍;
(8)干燥:出丸至脱盘上,送入干燥室,控制环境温度18-26℃,相对湿度20-35%,静态干燥24-36小时后,将胶囊出丸,拣丸(去除非正型丸)。
(9)包装成品:将正常型软胶囊装瓶、封装,即是用于抗化疗引起心肌毒性的鱼腥草挥发油护心胶囊成品。
实施例3:鱼腥草挥发油护心胶囊的抗化疗引起心肌毒性的减毒增效作用
1、实验材料
1.1材料与试剂
多柔比星(DOX:蒽环类常用化疗药物,用于建立心肌毒性动物模型) 深圳万乐药业有限公司;乙醇 分析纯,西陇化工股份有限公司、 美国Sigma公司;IL-1β试剂盒 武汉博士德公司。
实验动物:SD大鼠(180±20 g) 32只,雄性,南昌大学实验动物学科部。
1.2主要仪器设备
3K15-冷冻离心机 美国Sigma公司;SHP-150生化培养箱 上海森信实验仪器有限公司;AL-104电子天平 梅特勒-托利多仪器上海有限公司;Thermo 3001 Varioskan Flash全波长多功能酶标仪 美国Thermo公司;Milli-Q超纯水仪 美国Millipore公司;SKM型数显恒温电热套 山东鄄城华鲁电热仪器有限公司。
2 实验方法
2.1动物实验设计与分组
实验分组(每组8只):生理盐水组(正常组),DOX组(化疗组),DOX+挥发油100 mg/kg 组(化疗+鱼腥草挥发油组),DOX+鱼腥草挥发油护心胶囊100 mg/kg组(化疗+护心胶囊组)。挥发油母液:精密量取挥发油,以吐温-80作乳化剂,将其溶于生理盐水中,配成含0.5%吐温-80的鱼腥草挥发油。护心胶囊母液:精密量取等量挥发油,以吐温-80作乳化剂,将其溶于生理盐水中,重量比加入灵芝提取物和山楂提取物,配成护心胶囊母液,临用前将母液稀释至适宜的浓度。实验大鼠适应一周后,每天称重,第一天,正常组和化疗组灌胃生理盐水,其他组灌胃相对应的鱼腥草挥发油或护心胶囊液 ;第二天,化疗组,化疗+鱼腥草挥发油组及护心胶囊组腹腔注射DOX 20 mg/kg,正常组注射同剂量的生理盐水;第三天和第四天灌胃方式与第一天相同。第五天眼球取血,处死大鼠,取心脏组织。
2.2 血清中LDH活力的测定 对大鼠进行摘眼球取血,室温静置2小时,室温3000r/min离心10 min,收集血清,美国 Beckman 生化自动分析仪测定 LDH 活性。
使用LDH试剂盒测定LDH活力(U/L)。
2.3 血清中炎症因子IL-1β的测定 应用酶联免疫吸附测定(ELISA)血清中IL-1β的水平,具体操作细节参照试剂盒说明书进行,处理结束后,在酶标仪450nm处测定吸光度,根据标准曲线计算各组血清中IL-1β的水平 。
2.4数据处理 实验组数据用平均值±标准偏差(M±SD)的形式表示,采用SPSS23.0软件进行统计学分析。p<0.05时,表示与比较组存在显著差异,p<0.01表示与比较组差异极显著。
3 结果与分析
3.1鱼腥草挥发油护心胶囊对化疗引起心肌损伤的影响
乳酸脱氢酶(LDH)是一种常见的心肌损伤标志酶,本实验数据显示,与正常组数据显示,DOX处理组中LDH活力升高,表明化疗导致大鼠心肌损伤。然而,与化疗组相比,鱼腥草挥发油组LDH活力显著降低;同时,给予鱼腥草挥发油护心胶囊后,与化疗组相比,LDH的活力显著降低,且与鱼腥草挥发油组相比,LDH的活性有进一步下降的趋势。总之,本数据显示鱼腥草挥发油和鱼腥草挥发油护心胶囊组,均具有心肌保护作用可抑制化疗介导的心脏毒性,且护心胶囊组的心肌保护作用优于单独使用挥发油组的作用。
表2 大鼠体内LDH和IL-1β的含量
注:与正常组相比:# P <0.05,与模型组相比:* P <0.05,** P <0.01,与化疗+鱼腥草挥发油组相比:&p <0.05.
3.2 鱼腥草挥发油护心胶囊对心脏组织中炎症因子IL-1β的影响
临床数据显示DOX化疗处理后,可刺激机体免疫反应,释放炎症因子介导宿主免疫紊乱。IL-1β机体炎症反应过程中所释放的重要的炎性因子。与正常组相比,DOX组大鼠血清中的IL-1β含量明显上升,差异具有统计学意义。与DOX组相比,鱼腥草挥发油组和护心胶囊组中IL-1β含量显著下降;与鱼腥草挥发油组相比,护心胶囊组中IL-1β明显下降。上述结果表明,鱼腥草挥发油能抑制DOX介导的炎症因子IL-1β的升高,且护心胶囊组比单独使用挥发油组的效果更好。
总之,本产品在鱼腥草挥发油的基础上添加了灵芝和山楂提取物。灵芝具有扶正固本,可增强宿主的免疫调控能力,山楂具有健脾开胃、消食化滞之功效。动物实验数据显示,本产品与单独使用鱼腥草挥发油相比,其心肌保护作用更优,具有增效减毒作用。
Claims (4)
1.一种用于化疗减毒增效的鱼腥草挥发油护心胶囊,其特征在于:所述护心胶囊包括软胶囊内容物和囊壳,所述软胶囊内容物质量份数的物质组成:按重量计鱼腥草挥发油20~40份,辅料为山楂提取物10~20份、灵芝提取物5~10份和大豆油40~60份。
2.根据权利要求1所述的一种用化疗减毒增效的鱼腥草挥发油护心胶囊,其特征在于:所述囊壳包括明胶100份、甘油35-50份、水80-100份、山梨醇3-7份、棕氧化铁0.5-1份。
3.根据权利要求1或2所述的一种用于化疗减毒增效的鱼腥草挥发油护心胶囊的制备方法,其特征在于,包括以下步骤:
(1)鱼腥草干品、山楂、灵芝经粉碎机粉碎,过8-10目筛,得鱼腥草粉、山楂粉、灵芝粉;
(2)称取干燥鱼腥草粉,装入圆底烧瓶中,加10~40倍蒸馏水,混匀并浸泡10 h后,连接挥发油测定器与回流冷凝管;将整套装置放置于SKM型数显恒温电热套中,温度设置220℃,达到微沸状态,并保持微沸10 h,加热回流10 h后停止加热,收集鱼腥草挥发油;
(3)将山楂粉和灵芝粉加10~40倍蒸馏水经水煮2~4小时,煮两次,过滤合并两次滤液,旋转蒸发仪浓缩,减压真空干燥箱干燥,用超微粉碎机进行粉碎,过240-300目筛子,与鱼腥草挥发油及大豆油混匀得软胶囊内容物;
(4)胶液配制:按重量称取明胶100份、甘油35-50份、水80-100份、山梨醇3-7份、棕氧化铁0.5-1份,投入化胶罐中,搅拌融化明胶,控制真空度,减压抽真空,放入保温桶内,保温50-60℃,放置6-8小时,脱气,备用;
(5)制备鱼腥草挥发油软胶囊:将制备好的胶液和软胶囊内容物送入压丸机中,压制出软胶囊;
(6)定型:控制环境温度18-26℃,相对湿度20-35%,转笼定型1-3小时;
(7)洗丸:将上步得到的胶囊用95%以上的酒精洗去胶囊表面的油渍;
(8)干燥:出丸至脱盘上,送入干燥室,控制环境温度18-26℃,相对湿度20-35%,静态干燥24-36小时后,将胶囊出丸,拣丸;
(9)包装成品:将正常型软胶囊装瓶、封装,即是用于化疗减毒增效的鱼腥草挥发油护心胶囊成品。
4.根据权利要求3所述一种用于化疗减毒增效的鱼腥草挥发油护心胶囊,其特征在于:步骤(3)所述中软胶囊内容物即鱼腥草挥发油、山楂提取物、灵芝提取物和大豆油充分混匀。
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