CN110664860B - 一种增强免疫力的组合物及其制备方法 - Google Patents
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Abstract
本发明涉及保健食品技术领域,具体公开一种增强免疫力的组合物及其制备方法。所述增强免疫力的组合物包括如下重量份的组分:灵芝提取物5‑40份,沙棘提取物5‑35份,珍珠粉2‑32份,姜黄素0.5‑25份和雪莲培养物0.5‑20份。本发明根据中医学理论结合人体机能的特点,选择上述五味中药材进行配伍,并且通过功效成分与功能的关系,确定各药材的用量,使各组分之间协同作用,全方共呈气阴双补、阴阳并调、培元固本之功效,全面增强人体的免疫力,疗效显著,作用温和,尤其适合营养不良、劳累过度、免疫力低下的人群。且本发明产品未使用任何食品防腐剂,安全可靠。
Description
技术领域
本发明涉及保健食品技术领域,尤其涉及一种增强免疫力的组合物及其制备方法。
背景技术
免疫是人体的一种生理功能,人体依靠这种功能识别“自己”和“非己”成分,从而破坏和排斥进入人体的抗原物质,如病菌或人体本身所产生的损伤细胞和肿瘤细胞等,以维持人体的健康。免疫力功能是人体健康的基础,免疫功能失调和低下时,会降低机体对抗感染的能力,导致人体抗病力与自愈力的下降。而现代社会,生活节奏的加快,竞争压力的增大,使许多人处于紧张的工作和学习状态,机体长期处于这种紧张状态就会抑制免疫功能;同时,紧张的工作和学习让更多的人长期食用方便快餐,且经常熬夜,造成体内营养素的失衡,如果机体长期处于这种状态,会使机体免疫力下降,最终导致人体经常生病。而经常生病又会加重机体损耗,造成体质虚弱,精神萎靡,疲乏无力,如果不能及时恢复,则会进一步降低机体的免疫力,形成恶性循环。
近年来,随着人们健康意识的不断提升,越来越多的人希望通过保健食品来增强机体的免疫力,改善身体的健康状况。目前,虽然现用于增强免疫力的保健食品的种类繁多,但这些产品普遍存在成分复杂,效果不明显的缺陷,同时,由于成分复杂导致保健食品价格昂贵,无法长时间服用,不能达到持续、系统的增强免疫力的效果。因此,研制一种成分简单、有效、可长期服用的增强免疫力的保健食品具有十分重要的意义。
发明内容
针对现有技术中增强免疫力的保健食品成分复杂,且增强免疫力效果不明显的技术问题,本发明提供一种增强免疫力的组合物。
以及本发明还提供一种增强免疫力的组合物的制备方法。
以及本发明还提供一种增强免疫力的保健食品。
以及本发明还提供一种增强免疫力片。
以及本发明还提供一种增强免疫力片的制备方法。
为达到上述目的本发明实施例采用了如下的技术方案:
一种增强免疫力的组合物,所述组合物包括如下重量份的组分:灵芝提取物5-40份,沙棘提取物5-35份,珍珠粉2-32份,姜黄素0.5-25份和雪莲培养物0.5-20份。
相对于现有技术,本发明根据中医学理论结合人体机能的特点,选择灵芝、沙棘、珍珠粉、姜黄素和雪莲培养物五味中药材进行配伍,并且通过功效成分与功能的关系,确定各药材的用量,使各组分之间协同作用,沙棘、姜黄素、雪莲培养物协同,不但可以增强机体心脏的收缩和舒张功能,促进毛细血管血液循环,提高机体免疫力,同时还能改善机体的消化系统功能,促进机体对组合物中活性组分的吸收利用,提高各活性组分的生物利用度;灵芝中丰富的有机锗和沙棘黄酮配合,可以改善大脑的兴奋与抑制过程,增强机体免疫能力,对大脑的神经系统具有明显的保护和协调作用;珍珠粉不但可以促进淋巴器官的生长发育,提高人体免疫系统中的T、B淋巴细胞的活性,增强免疫抗病能力,同时,还能改善视神经功能,且还能调和雪莲培养物的性热,最大限度发挥雪莲培养物增强免疫力的功效,使组方补而不燥,全方共呈气阴双补、阴阳并调、培元固本之功效,适合各类人群长期服用。且本发明产品未使用任何食品防腐剂,原料均为保健食品可用原料,安全可靠。
灵芝,性温,可补心、肝、脾、肺、肾五脏之气,具滋补强壮,扶正固本之功效,主含氨基酸、多肽、蛋白质、真菌溶菌酶,多糖、麦角甾醇、三萜类、生物碱、维生素B2、维生素C等。灵芝中的三萜类物质能够促进肝脏对药物、毒物的代谢,提高肝脏的解毒能力,促进机体的新陈代谢;灵芝中的多糖和多肽可促进机体核酸和蛋白质的合成,同时,灵芝中的多糖和微量元素,如锌、铁、锰以及维生素可提高机体的超氧化物歧化酶的活性,加快清除体内的自由基,加速细胞的再生;灵芝中的真菌溶菌酶还能增强机体对细菌的抵抗能力;灵芝中含有的特殊的有机锗可以提高人体血液含氧量,加快新陈代谢,同时还能与人体内残留的重金属离子等有毒物质结合成锗化合物,随大小便排除体外,达到净化血液,促进新陈代谢,防止老化的作用。灵芝中的多糖、多肽以及富含的微量元素和维生素,从加快新陈代谢、促进新蛋白和核酸合成,以及加快有毒物质、衰老或损伤细胞的清除两方面共同作用,加快新老交替,使机体细胞处于青春状态,从而增强机体的免疫力。
沙棘,性温,味酸,含有多种维生素、微量元素、亚油素、沙棘黄酮、超氧化物等活性物质和人体所需的各种氨基酸。沙棘中富含的维生素E、胡萝卜素、类胡萝卜素、β-谷甾醇、不饱和脂肪酸等,可促进机体新陈代谢,有利于损伤组织的恢复,增强炎症中心的抗炎作用,以及明显的促进溃疡愈合作用;沙棘黄酮可以增强心脏的收缩和舒张功能,有明显的抗心肌缺血和抗心律失常的功效,同时还有消炎和促进毛细血管血液循环的能力,对于长期熬夜,心里压力大的人群十分有利;同时,沙棘中富含的氨基酸和有机酸可以促进胃酸、胃液分泌,促进机体对本发明中灵芝、雪莲等其他组分中活性组分的吸收利用,提高活性组分的生物利用度。灵芝中的有机锗与沙棘黄酮配合,可改善大脑的兴奋与抑制过程,增强机体免疫能力,对大脑的神经系统具有明显的保护和协调作用,增强机体在特殊环境(如低温、缺氧)中的耐寒能力和耐缺氧能力,增强机体活力。
珍珠粉,含有丰富的氨基酸,如亮氨酸、蛋氨酸、丙氨酸等18种氨基酸,可促进淋巴器官的生长发育,从而有效提高人体免疫系统中的T、B淋巴细胞的活性,增强免疫抗病能力;其含有的独特的牛磺酸具有调节中枢神经系统的作用,为增强机体的免疫力提供多方面的功能协调,还可以增强心肌的收缩力,防治心脏功能衰竭;珍珠粉中的硒可以强化视觉神经的传导,改善视力状况;微量元素中的锌可促进视神经的轴浆运输,改善视网膜及视网膜色素上皮细胞的代谢与功能。珍珠粉中含有的微量元素如硒、锗、锰、铜、锌等,可提高沙棘中高含量的超氧化物歧化酶的活性,促进机体细胞的再生。
姜黄素,味辛,性温,归脾、肝经,能够抑制多种细菌的生长,具有优异的抗炎作用,姜黄素在抗氧化过程中可以产生稳定性很好的醌类物质,其可作用一种很好的抗氧化剂,具有较强的抗自由基、抗脂质过氧化和稳定细胞膜的作用。本发明中姜黄素和沙棘、雪莲等协同作用,可抑制氧自由基的生成,增加谷胱甘肽的水平,进而保护机体组织器官免受侵害,增强机体对疾病的抵抗能力。同时,姜黄素和沙棘共同作用,还能改善机体消化系统的功能,增强机体对各组分活性物质的吸收,提高活性物质的生物利用度,提高组合物增强免疫力的功效。
雪莲培养物,性大热,入肝脾肾三经,能补阴益阳,营养神经,含有丰富的黄酮类、多糖类、绿原酸、紫丁香苷和生物碱等活性物质,还含有18种氨基酸,且不仅含有与天然雪莲相近的活性成分,且亦具有与之相当的多方面的药理活性。雪莲培养物中的多糖对生物体内超氧阴离子自由基具有明显的清除作用,可延缓衰老的产生;雪莲培养物中的总黄酮,具有免疫调节的作用,可增强机体的免疫力;雪莲培养物中还含有独特的黄酮类和雪莲内脂可有效清除氧自由基,防止染色体畸变,具有优异的防辐射能力;雪莲果中富含纤维,可以有效促进肠道蠕动、清除肠道垃圾,改善便秘、防止腹泻,改善肠胃功能;雪莲中含有的紫丁香苷与沙棘、灵芝中的维生素B协同作用,可促进机体促进钙、镁、铁等矿物质吸收,提高机体新陈代谢水平,增强免疫力和抗病力。
本发明中雪莲为大热物质,不适合阴虚火旺的人群长期服用,配以特定含量的珍珠粉,可调和雪莲的性热,使组方补而不燥,适合各种人群长期服用。本发明中灵芝、沙棘、珍珠粉和姜黄素可加强雪莲培养物的功效,上五味药合用,补元气,养五脏,调节人体气血、阴阳的平衡,平衡人体营养,加速机体新陈代谢,延缓细胞衰老,全面增强人体的免疫力,疗效显著,作用温和,尤其适合营养不良、年老体衰、劳累过度、精神不佳、免疫力低下的人群。
优选的,所述组合物包括如下重量份的组分:灵芝提取物10-30份,沙棘提取物8-30份,珍珠粉5-25份,姜黄素2-20份和雪莲培养物2-15份。
更优选的,所述组合物包括如下重量份的组分:灵芝提取物20份,沙棘提取物18份,珍珠粉10份,姜黄素8份和雪莲培养物8份。
优选的,所述灵芝提取物的制备方法为:将灵芝粉碎,得灵芝粗粉,向所述灵芝粗粉中加入其质量4-8倍的水,于90-100℃提取3-4次,每次提取时间为1-3h,过滤,合并滤液,减压浓缩,醇沉,真空干燥,得灵芝提取物。
优选的,所述沙棘提取物的制备方法为:将沙棘原料粉碎,加入超临界萃取仪中,通入二氧化碳介质进行萃取,得沙棘粗粉,向所述沙棘粗粉中加入其质量5-15倍的质量浓度为65-75%的乙醇溶液,浸泡1-3h后,加热回流提取 3-4次,每次提取时间为1-3h,过滤,合并滤液,减压浓缩,真空干燥,得沙棘提取物。
灵芝和沙棘含有多种有效成分,如黄酮、总皂苷、多糖等,采用上述制备方法可将灵芝和沙棘的活性物质充分提取出来,从而提高本发明组合物的增强免疫力的作用。
可选的,灵芝提取物制备方法中减压浓缩的条件为:温度为50-75℃,真空度为0.06-0.1MPa,将滤液浓缩至比重为1.10-1.13。真空干燥的温度为50- 60℃,真空度为0.04-0.06MPa。
可选的,沙棘提取物制备方法中减压浓缩的条件为:温度为55-65℃,真空度为0.06-0.08MPa,将滤液浓缩至比重为1.02-1.06;真空干燥的温度60- 70℃,真空度为0.06-0.08MPa。
优选的,沙棘提取物制备方法中超临界萃取的压力为15-30MPa,温度为 35-40℃,萃取时间为3-4h,二氧化碳流速为25L/h。
沙棘中活性成分含量低,而且还含有许多其他杂质,如淀粉、糖类、油脂等,通过超临界萃取法可将沙棘中的大量杂质进行脱除,然后再采用乙醇溶液进行提取,可将沙棘中的活性有效成分选择性地萃取出来,提高沙棘的增强免疫力的药效。
本发明还提供一种增强免疫力的组合物的制备方法,至少包括以下步骤:
按照设计配比称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素和雪莲培养物,混合均匀,得增强免疫力的组合物。
本发明还提供一种增强免疫力的保健食品,包括上述的增强免疫力的组合物。
将增强免疫力的组合物制成保健食品后,更适宜食用者服用和携带,便于长期食用,达到增强免疫力的效果。
优选的,所述增强免疫力的保健食品的剂型为粉剂、颗粒剂、胶囊剂或片剂。
本发明具有增强免疫力的组合物在制备成保健食品时,除了包括有效的药物成分之外,还应包括制剂学上允许使用的载体如赋形剂和辅料。这些载体的作用是有助于有效成分加工成制剂,并且运载有效成分到体内作用部位。
本发明还提供一种增强免疫力片,包括上述的增强免疫力的组合物,还包括如下重量份数的原料:乳糖10-30份,微晶纤维素10-30份,聚维酮K30 0.5-6份,硬脂酸镁0.2-3份和二氧化硅0.2-3份。
本发明还提供一种增强免疫力片的制备方法,至少包括以下步骤:
按照设计配比称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素、雪莲培养物、乳糖、微晶纤维素、聚维酮K30、硬脂酸镁和二氧化硅,分别过60- 80目筛,混合均匀,得总混粉,将所述总混粉压片,得增强免疫力片。
该制备方法工序通过混合、压片,使所得产品稳定性好,方便食用,且制备工艺无复杂工序,方法简单,适合工业化生产。
优选的,所述增强免疫力片的制备方法还包括以下步骤:
取薄膜包衣预混剂,用纯化水溶解,配制成固含量为17-19%的包衣液,将所述增强免疫力片包衣,包衣至素片增重1.8-2.2%。
含有本发明增强免疫力的片剂还可以包衣,以提供达到延长药效作用的剂型。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合实施例,对本发明进行进一步详细说明。应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。
实施例1
本发明实施例提供一种增强免疫力片,包括如下重量份的组分:
灵芝提取物5份,沙棘提取物35份,珍珠粉2份,姜黄素25份,雪莲培养物0.5份,乳糖20份,微晶纤维素10份,聚维酮K30 3份,硬脂酸镁0.2 份和二氧化硅3份。
上述增强免疫力的片剂的制备步骤如下:
步骤a、将灵芝粉碎,得灵芝粗粉,向所述灵芝粗粉中加入其质量8倍的水,于90℃提取3h,过滤,得第一滤液和第一滤渣;向所述第一滤渣中加入 5倍量的水,于90℃提取3h,得第二滤液和第二滤渣;向所述第二滤渣中加入4倍量的水,于90℃提取2h,得第三滤液和第三滤渣;向所述第三滤渣中加入4倍量的水,于90℃提取1h,得第四滤液;合并上述滤液,于60℃,真空度为0.08MPa的条件下减压浓缩至比重为1.10-1.13,加入浓缩物质量3倍量的质量浓度为95%的乙醇溶液进行醇沉,离心,于55℃、真空度为 0.05MPa的条件下减压干燥,粉碎,过80目筛,得灵芝提取物;
步骤b、将沙棘原料粉碎至20-40目,加入超临界萃取仪中,通入二氧化碳介质进行萃取,得沙棘粗粉,向所述沙棘粗粉中加入其质量15倍的质量浓度为65%的乙醇溶液,浸泡1h后,加热回流2h,过滤,得第一滤液和第一滤渣;向所述滤渣中加入10倍量的上述乙醇溶液,加热回流提取2h,过滤,得第二滤液和第二滤渣;向所述第二滤渣中加入8倍量的上述乙醇溶液,热回流提取2h,过滤,得第三滤液和第三滤渣;向所述第三滤渣中加入5倍量的上述乙醇溶液,热回流提取2h,过滤,得第四滤液;合并上述滤液,于60℃、真空度为0.07MPa的条件下减压浓缩至比重为1.02-1.06,于65℃、真空度为 0.07MPa的条件下进行干燥,粉碎,得沙棘提取物;
其中,超临界萃取的条件为:超临界萃取的压力为15MPa,温度为 40℃,萃取时间为4h,二氧化碳流速为25L/h。
步骤c、按照设计配比称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素、雪莲培养物、乳糖、微晶纤维素、聚维酮K30、硬脂酸镁和二氧化硅,分别过60目筛,混合均匀,得总混粉,将所述总混粉压片,得增强免疫力片。
本实施例制备的片剂片重0.5g,服用方法为每天一次,每次2片。
实施例2
本发明实施例提供一种增强免疫力片,包括如下重量份的组分:
灵芝提取物40份,沙棘提取物5份,珍珠粉32份,姜黄素0.5份,雪莲培养物20份,乳糖10份,微晶纤维素30份,聚维酮K30 6份,硬脂酸镁3 份和二氧化硅2份。
上述增强免疫力的片剂的制备步骤如下:
步骤a、将灵芝粉碎,得灵芝粗粉,向所述灵芝粗粉中加入其质量7倍的水,于95℃提取3h,过滤,得第一滤液和第一滤渣;向所述第一滤渣中加入 5倍量的水,于95℃提取2h,得第二滤液和第二滤渣;向所述第二滤渣中加入4倍量的水,于95℃提取2h,得第三滤液;合并上述滤液,于60℃,真空度为0.08MPa的条件下减压浓缩至比重为1.10-1.13,加入浓缩物质量3倍量的质量浓度为95%的乙醇溶液进行醇沉,离心,于55℃、真空度为0.05MPa 的条件下减压干燥,粉碎,过80目筛,得灵芝提取物;
步骤b、将沙棘原料粉碎至20-40目,加入超临界萃取仪中,通入二氧化碳介质进行萃取,得沙棘粗粉,向所述沙棘粗粉中加入其质量14倍的质量浓度为75%的乙醇溶液,浸泡3h后,加热回流3h,过滤,得第一滤液和第一滤渣;向所述滤渣中加入12倍量的上述乙醇溶液,加热回流提取2h,过滤,得第二滤液和第二滤渣;向所述第二滤渣中加入10倍量的上述乙醇溶液,热回流提取2h,过滤,得第三滤液;合并上述滤液,于60℃、真空度为0.07MPa 的条件下减压浓缩至比重为1.02-1.06,于65℃、真空度为0.07MPa的条件下进行干燥,粉碎,得沙棘提取物;
其中,超临界萃取的条件为:超临界萃取的压力为30MPa,温度为 35℃,萃取时间为3h,二氧化碳流速为25L/h。
步骤c、按照设计配比称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素、雪莲培养物、乳糖、微晶纤维素、聚维酮K30、硬脂酸镁和二氧化硅,分别过80目筛,混合均匀,得总混粉,将所述总混粉压片,得增强免疫力片。
本实施例制备的片剂片重0.5g,服用方法为每天一次,每次2片。
实施例3
本发明实施例提供一种增强免疫力片,包括如下重量份的组分:
灵芝提取物20份,沙棘提取物18份,珍珠粉10份,姜黄素8份和雪莲培养物8份,乳糖30份,微晶纤维素20份,聚维酮K30 0.5份,硬脂酸镁 1.5份,二氧化硅0.2份和包衣预混剂2份。
上述增强免疫力的片剂的制备步骤如下:
步骤a、将灵芝粉碎,得灵芝粗粉,向所述灵芝粗粉中加入其质量7倍的水,于100℃提取2h,过滤,得第一滤液和第一滤渣;向所述第一滤渣中加入 6倍量的水,于100℃提取2h,得第二滤液和第二滤渣;向所述第二滤渣中加入5倍量的水,于100℃提取2h,得第三滤液;合并上述滤液,于60℃,真空度为0.08MPa的条件下减压浓缩至比重为1.10-1.13,加入浓缩物质量3倍量的质量浓度为95%的乙醇溶液进行醇沉,离心,于55℃、真空度为0.05MPa的条件下减压干燥,粉碎,过80目筛,得灵芝提取物;
步骤b、将沙棘原料粉碎至20-40目,加入超临界萃取仪中,通入二氧化碳介质进行萃取,得沙棘粗粉,向所述沙棘粗粉中加入其质量12倍的质量浓度为70%的乙醇溶液,浸泡2h后,加热回流2h,过滤,得第一滤液和第一滤渣;向所述滤渣中加入8倍量的上述乙醇溶液,加热回流提取1.5h,过滤,得第二滤液和第二滤渣;向所述第二滤渣中加入6倍量的上述乙醇溶液,热回流提取1h,过滤,得第三滤液;合并上述滤液,于60℃、真空度为0.07MPa的条件下减压浓缩至比重为1.02-1.06,于65℃、真空度为0.07MPa的条件下进行干燥,粉碎,得沙棘提取物;
其中,超临界萃取的条件为:超临界萃取的压力为25MPa,温度为 38℃,萃取时间为3.5h,二氧化碳流速为25L/h。
步骤c、按照设计配比称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素、雪莲培养物、乳糖、微晶纤维素、聚维酮K30、硬脂酸镁和二氧化硅,分别过60目筛,混合均匀,得总混粉,将所述总混粉压片,得素片。
步骤d:取薄膜包衣预混剂,用纯化水溶解,配制成固含量为18%的包衣液,将素片包衣,包衣至素片增重2%,得增强免疫力片。
本实施例制备的片剂片重0.5g,服用方法为每天一次,每次2片。
本发明中增强免疫力的组合物还可选择保健食品领域常规的辅料按照常规的工艺制备为粉末剂、颗粒剂和胶囊剂。
增强免疫力功能试验
试验对象:
本试验对象为SPF级昆明种雄性小鼠160只,体重18-22g。
剂量设计:
将实施例3制备的增强免疫力片设计为0.50g/Kg.BW,0.34g/Kg.BW, 0.17g/Kg.BW三个剂量组,其剂量分别相当于人体推荐用量的30倍、20倍、 10倍,另设计一个阴性对照组(蒸馏水)。
样品配制:分别称取实施例3制备的增强免疫力片1.0g、0.68g、0.34g分别加蒸馏水至40mL,混合均匀,配制成浓度为2.50%、1.70%和0.85%的混悬液,备用。
试验方法:
将雄性小鼠160只分为4个试验组,每组用40只小鼠并随机分成4组,各试验组均设三个剂量组(1个中剂量组、1个高剂量组,1个低剂量组)和1 个阴性对照组,每组10只。分别用于第一组小鼠迟发型变态反应(DTH)测定,第二组抗体生成细胞检测、血清溶血素的测定,第三组小鼠碳廓清试验和第四组小鼠腹腔巨噬细胞吞噬鸡红细胞试验。按上述剂量每天灌胃给予受试样品一次,阴性对照组给予等容量的蒸馏水,灌胃容量为20mL/Kg.BW,连续30天。
试验方法及结果:
1.小鼠迟发型变态反应(DTH)测定
给受试样品第26天,腹腔注射2%(v/v)SRBC 0.2mL/只,致敏动物,致敏后第四天每鼠左后足跖皮下注射20%(v/v)SRBC 20μL/只进行攻击。并于攻击前和攻击后24h分别测量每只鼠左后足跖同一部位厚度,同一部位测三次,取平均值。计算攻击前、后的足跖厚度差值,各剂量组结果与阴性对照组比较进行方差分析。结果如表1所示。
表1
注:各剂量组与阴性对照组比,*表示P<0.05。
由上述结果可以看出,各剂量组小鼠各期体重及增重与阴性对照组比较,均无显著性差异(P>0.05);高剂量组能增强小鼠对SBRC诱发的DTH反应,与阴性对照组比,差异有显著性(P<0.05),中、低剂量组与阴性对照组比无显著差异。
2.抗体生成细胞检测试验
经腹腔注射2%(v/v)的SRBC0.2mL/只,于免疫4d后的小鼠颈椎脱臼处死,取出脾脏,放在盛装有Hank’s液的小平皿内,轻轻磨碎脾脏,制成细胞悬液,经200目筛网过滤,离心(1000r/min)10min,用Hank’s液洗2 遍,最后将细胞悬浮在5mLRPMI1640培养液中,计数细胞,并将细胞浓度调整为5×106个/mL。
空斑的测定:将表层培养基(lg琼脂糖加双蒸水至l00mL)加热溶解后,放入45-50℃水浴保温,与等量pH7.2-7.4、2倍浓度的Hank’s液混合,分装小试管,每管0.5mL,再向管内加50μ110%SRBC(v/v,用SA缓冲液配制), 20μL脾细胞悬液(5×106个/mL),迅速混匀,倾倒于己刷琼脂糖薄层的玻片上,做平行片,待琼脂凝固后,将玻片水平扣放在片架上,放入二氧化碳培养箱中孵育I.5h,然后用SA缓冲液稀释的补体(l:8)加入玻片架凹槽内,继续温育I.5h后,计数溶血空斑数。结果如表2所示。
表2
注:各剂量组与阴性对照组比,*表示P<0.05。
由上述结果可以看出,各剂量组小鼠各期体重及增重与阴性对照组比较,均无显著性差异(P>0.05);高剂量组与阴性对照组比较,差异有显著性 (P<0.05)。结果表明,该样品高剂量组能增强其抗体生成细胞数量。
3.血清溶血素的测定
给受试样品第26天每鼠腹腔注射2%SRBC 0.2mL/只,于免疫后第四天后摘眼球采血。分离血清。用生理盐水将血清倍比稀释,将不同稀释度的血清分别置于微量血凝板内,每孔100μL,再加入100μL0.5%(v/v)的SRBC悬液,混匀,装入湿润的平盘内加盖,于37℃温箱孵育3h进行血清溶血素测定。统计血球凝集度,计算相应抗体积数。结果如表3所示。
表3
组别 | 血清溶血素(抗体积数) | P值 |
高剂量组 | 180.3±13.9<sup>*</sup> | 0.041 |
中剂量组 | 171.6±25.5 | 0.268 |
低剂量组 | 160.0±19.7 | 0.968 |
阴性对照组 | 156.6±22.6 | - |
注:各剂量组与阴性对照组比,*表示P<0.05。
由上述结果可以看出,高剂量组与阴性对照组比较,差异有显著性 (P<0.05)。结果表明,该样品高剂量组能增强其血清溶血素水平。
4.碳廓清试验
各组小鼠尾静脉注射印度墨汁(用生理盐水4.0倍稀释)l0rnL/kg.BW。每鼠在注射墨汁后2min和l0min分别通过眼内眦静脉丛各取血20μL,并迅速加入到0.1%碳酸钠溶液2mL中并摇匀。以碳酸钠溶液作空白对照,用紫外可见分光光度计600nm波长处测光密度值(OD)。将小鼠处死后取肝脏和脾脏,用滤纸吸干脏器表面血污,分别称重。按公式计算吞噬指数(a),结果如表4所示。
表4
注:各剂量组与阴性对照组比,*表示P<0.05。
由上述结果可以看出,各剂量组小鼠各期体重及增重与阴性对照组比较,均无显著性差异(P>0.05);高剂量组与阴性对照组比较,差异有显著性 (P<0.05)。结果表明,该样品高剂量组能提高小鼠碳廓清吞噬指数。
5.小鼠腹腔巨噬细胞吞噬鸡红细胞试验
采用半体内法。各组小鼠均腹腔注射20%(v/v)鸡红细胞悬液lmL/只,间隔30分钟颈推脱臼处死小鼠,经腹腔注入生理盐水2mL/只。转动鼠板lmin 后吸出腹腔洗液lmL平均分滴于2片载玻片上,放入垫有湿纱布的搪瓷盒内,37℃温育30min。孵毕,再用生理盐水漂洗、晾干,以1:1的丙酮甲醇溶液固定,4%(v/v)Giemsa-磷酸缓冲液染色,再用蒸馏水漂洗晾干。显微镜油镜下观察100个巨噬细胞,计数吞噬鸡红细胞的巨噬细胞数和巨噬细胞吞噬的鸡红细胞总数及其被消化的程度,按公式求出吞噬率、吞噬指数,结果如表 5所示。
表5
注:各剂量组与阴性对照组比,*表示P<0.05。
由上述结果可以看出,各剂量组小鼠各期体重及增重与阴性对照组比较,均无显著性差异(P>0.05);高剂量组小鼠巨噬细胞吞噬鸡红细胞吞噬百分率和吞噬指数与阴性对照组相比,差异均有显著性(P<0.05)。
实施例4
本发明实施例提供一种增强免疫力片,包括如下重量份的组分:
灵芝提取物10份,沙棘提取物30份,珍珠粉6份,姜黄素20份和雪莲培养物2份,乳糖15份,微晶纤维素25份,聚维酮K30 5份,硬脂酸镁0.5 份,二氧化硅2份和包衣预混剂3份。
上述增强免疫力片的制备方法与实施例3相同,此处不再赘述。
实施例5
本发明实施例提供一种增强免疫力片,包括如下重量份的组分:
灵芝提取物30份,沙棘提取物8份,珍珠粉20份,姜黄素2份和雪莲培养物15份,乳糖30份,微晶纤维素15份,聚维酮K30 1份,硬脂酸镁2 份,二氧化硅3份和包衣预混剂2.5份。
上述增强免疫力片的制备方法与实施例3相同,此处不再赘述。
对比例1
本对比例提供一种增强免疫力片,原料按重量份数计包括如下组分:丹参提取物20份,沙棘提取物18份,珍珠粉10份,姜黄素8份,雪莲培养物8 份,乳糖30份,微晶纤维素20份,聚维酮K30 0.5份,硬脂酸镁1.5份,二氧化硅0.2份和包衣预混剂2份。
对比例2
本对比例提供一种增强免疫力片,原料按重量份数计包括如下组分:灵芝提取物20份,沙棘提取物18份,珍珠粉10份,姜黄素8份,雪莲培养物8 份,西洋参9份,乳糖30份,微晶纤维素20份,聚维酮K30 0.5份,硬脂酸镁1.5份,二氧化硅0.2份和包衣预混剂2份。
按照上述实施例3中的增强免疫力试验的测试方法,测试实施例1-5以及对比例1和对比例2中增强免疫力片的溶血空斑数、血清溶血素、碳廓清吞噬指数a以及吞噬鸡红细胞的吞噬率,按照高剂量组的试验方法进行。将实施例 1-5、阴性对照组、对比例1-2的测试结果汇总,如表 6 所示。
表 6
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换或改进等,均应包括在本发明的保护范围之内。
Claims (8)
1.一种增强免疫力的组合物,其特征在于,所述组合物由如下重量份的组分组成:灵芝提取物5-40份,沙棘提取物5-35份,珍珠粉2-32份,姜黄素0.5-25份和雪莲培养物0.5-20份;
所述灵芝提取物的制备方法为:
将灵芝粉碎,得灵芝粗粉,向所述灵芝粗粉中加入其质量4-8倍的水,于90-100℃提取3-4次,每次提取时间为1-3h,过滤,合并滤液,减压浓缩,醇沉,干燥,得灵芝提取物;
所述沙棘提取物的制备方法为:
将沙棘原料粉碎,加入超临界萃取仪中,通入二氧化碳介质进行萃取,得沙棘粗粉,向所述沙棘粗粉中加入其质量5-15倍的质量浓度为65-75%的乙醇溶液,浸泡1-3h后,加热回流提取3-4次,每次提取时间为1-3h,过滤,合并滤液,减压浓缩,真空干燥,得沙棘提取物。
2.如权利要求1所述的增强免疫力的组合物,其特征在于,所述组合物由如下重量份的组分组成:灵芝提取物10-30份,沙棘提取物8-30份,珍珠粉5-25份,姜黄素2-20份和雪莲培养物2-15份。
3.如权利要求1所述增强免疫力的组合物,其特征在于,所述组合物由如下重量份的组分组成:灵芝提取物20份,沙棘提取物18份,珍珠粉10份,姜黄素8份和雪莲培养物8份。
4.如权利要求1所述的增强免疫力的组合物,其特征在于,超临界萃取的压力为15-30MPa,温度为35-40℃,萃取时间为3-4h,二氧化碳流速为25L/h。
5.一种增强免疫力的保健食品,其特征在于,包括权利要求1-4任一项所述的增强免疫力的组合物。
6.如权利要求5所述的增强免疫力的保健食品,其特征在于:其剂型为粉剂、颗粒剂、胶囊剂或片剂。
7.一种增强免疫力片,其特征在于,所述增强免疫力片包括权利要求1-4任一项所述的增强免疫力的组合物,还包括如下重量份数的原料:乳糖10-30份,微晶纤维素10-30份,聚维酮K30 0.5-6份,硬脂酸镁0.2-3份和二氧化硅0.2-3份。
8.权利要求7所述的增强免疫力片的制备方法,其特征在于,至少包括以下步骤:
按照重量份数称取所述灵芝提取物、沙棘提取物、珍珠粉、姜黄素、雪莲培养物、乳糖、微晶纤维素、聚维酮K30、硬脂酸镁和二氧化硅,分别过60-80目筛,混合均匀,得总混粉,将所述总混粉压片,得增强免疫力片。
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