CN114917260A - 一种富硒香菇提取物的制备方法及其应用 - Google Patents
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Abstract
本发明公开了一种富硒香菇提取物的制备方法及其应用,本发明的制备方法包含:(1)选材:所用富硒香菇为子实体,子实体干品中总硒含量不低于50mg/kg,其中有机硒含量不低于90%;(2)富硒香菇提取物制备:富硒香菇子实体低温烘干,干粉粉碎,加5~20倍体积水浸泡,超声提取10~30min,减压浓缩,冻干,得水提物干粉。利用本方法制备出的提取物,对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)细胞系有显著的抗增殖活性。本发明为该提取物的抗癌或辅助治疗作用研究奠定了一定的基础。
Description
技术领域
本发明属于生物提取物技术领域,特别涉及一种富硒香菇提取物的制备方法及其应用。
背景技术
香菇又名冬菇、香蕈等,是世界第二大食用菌,我国第一大食用菌,我国2020年产量达1188万吨。香菇肉质肥厚细嫩,味道鲜美,营养丰富,含有抗肿瘤、降血脂等功效成分,是广受欢迎的食用菌。
硒是人体必需的微量元素,被称为微量元素中的“生命奇效元素”。硒是人体多种酶的重要组成部分,包括谷胱甘肽过氧化物酶、脱碘酶、硫氧还蛋白还原酶、硒代磷酸合成酶等。
硒的生理活性态是有机硒,无机硒毒性强、在体内不易吸收,必须要转化为有机硒才能发挥其生理活性。香菇是转化合成有机硒的良好载体。富硒香菇具有抗癌、调节免疫等多方面作用,其效果优于普通香菇。
现有技术中,公开号CN 112999241 A《一种香菇多糖纳米硒及制备方法以及其在治疗恶性胸腔积液中的应用》提供了一种香菇多糖纳米硒的制备方法以及在制备抗癌药物和/或防治恶性胸腔积液(MPE)的药物中的应用,虽然该方法取得了一定的效果,但是针对的仅是治疗恶性胸腔积液,在利用香菇抗癌特别是富硒香菇抗癌方面仍有很大的研究空间。
发明内容
本发明要解决的技术问题是针对现有技术的不足,提供一种富硒香菇提取物制备方法及其应用,丰富抗癌或辅助治疗作用产品。
本发明的技术方案:一种富硒香菇提取物的制备方法,所用富硒香菇为子实体,子实体干品中总硒含量不低于50mg/kg,其中有机硒含量不低于90%。
富硒香菇提取物制备方法,其步骤是,富硒香菇子实体低温烘干,干粉粉碎,加5~20倍体积水浸泡,超声提取10~30min,减压浓缩,冻干。
将提取物冻干粉,配成梯度药物,体外试验检测其对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)等癌细胞的抗癌效果,计算其IC50。
本发明的有益效果:本发明公开的一种富硒香菇提取物的制备方法,提取工艺简单,而且获得的提取物具有良好的抗癌活性。本发明富硒香菇水提取物对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)细胞系有显著的抗增殖活性,IC50分别为308.2 μg/mL,293.6 μg/mL,309.5 μg/mL,263.5 μg/mL,292.122 μg/mL。尤其对肝癌(HepG2)、宫颈癌(Hela)、白血病(K562)作用效果非常明显。本发明为该富硒香菇提取物的抗癌或辅助治疗作用研究奠定了一定的基础。
附图说明
图1是富硒香菇水提物对人前列腺癌(LNCaP)细胞抑制率影响图;
图2是富硒香菇水提物对肝癌(HepG2)细胞抑制率影响图;
图3是富硒香菇水提物对肺癌(A549)细胞抑制率影响图;
图4是富硒香菇水提物对宫颈癌(Hela)细胞抑制率影响图;
图5是富硒香菇水提物对白血病(K562)细胞抑制率影响图。
具体实施方式
下面对本发明的具体实施方式作进一步说明。在此需要说明的是,对于这些实施方式的说明用于帮助理解本发明,但并不构成对本发明的限定。此外,下面所描述的本发明各个实施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
下述实验例中所使用的试验方法如无特殊说明,均为常规方法;所使用的材料、试剂等,如无特殊说明,为可从商业途径得到的试剂和材料。
实施例1
一种富硒香菇提取物的制备,包括以下步骤:
(1)将选用的新鲜富硒香菇子实体(其子实体干品中总硒含量不低于50mg/kg,其中有机硒含量不低于90%),在50-80℃低温烘干,粉碎过60目筛。
(2)取粉碎的富硒香菇子实体干粉100g,加10倍体积水1L浸泡1h,超声提取30min,减压浓缩,冻干,得水提物冻干粉。
实施例2
富硒香菇提取物提取物的体外抗增殖活性实验
(1)活性实验中用到的细胞株,试剂以及仪器
人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)。DMEM培养液(GIBCO),胎牛血清(四季青),胰蛋白酶(GIBCO),DMSO(分析纯),MTS(Amresco),96孔板,细胞培养箱(Thermo),酶标仪(Thermo),超净工作台。
(2)肿瘤细胞培养方法
基础培养基1640+10BS(胎牛血清),培养条件为37℃,5%CO2浓度及饱和湿度;当细胞生长至对数期时,用胰蛋白酶消化细胞,洗涤吹散细胞,以每孔5000个细胞接种于96孔板,每孔体积100 µL。细胞提前24小时接种培养,以备MTT实验。
(3)细胞增殖实验(MTT法)
将富硒香菇水提物干粉溶于DMS0溶液,配制浓度为4 mg/mL的母液。待细胞贴壁后,随机分为对照组和富硒香菇水提物不同质量浓度组(终浓度为25、50、100、200、400 µg/mL),另设不含细胞只含完全培养基的空白组,每组设置5个复孔。对照组加入含0.1%DMSO的完全培养基100 µL,富硒香菇水提物各质量浓度组分别加入含相应药物的完全培养基100 µL,培养24 h 时,向各组细胞加入 MTT 溶液 20 µL,于37℃条件下孵育4 h后,以2500 r/min离心15 min,弃上清;沉淀中加入 DMSO 150 µL,采用酶标仪于 490 nm波长处测定各孔吸光度(OD),计算细胞存活率。
细胞存活率(%)=(OD 实验组-OD 空白组)/(OD对照组-OD 空白组)×100%
(4)实验结果见表1。从表中可以看出:富硒香菇水提物对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)均显示出显著的细胞增殖抑制活性。
表1 富硒香菇提取物对五种细胞株的抗增殖活性数据
富硒香菇水提物对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)细胞抑制率影响图,分别见图1、图2、图3、图4、图5。
富硒香菇水提取物对人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)细胞系有显著的抗增殖活性,其IC50分别为308.2 μg/mL,293.6 μg/mL,309.5 μg/mL,263.5 μg/mL,292.122 μg/mL。尤其对肝癌(HepG2)、宫颈癌(Hela)、白血病(K562)作用效果非常明显。本发明为该富硒香菇提取物的抗癌或辅助治疗作用研究奠定了一定的基础。
Claims (4)
1.一种富硒香菇提取物的制备方法,其特征在于,包括以下步骤:
1)选材:所用富硒香菇为子实体,子实体干品中总硒含量不低于50mg/kg,其中有机硒含量不低于90%;
2)富硒香菇提取物制备:富硒香菇子实体低温烘干,干粉粉碎,加5~20倍体积水浸泡,超声提取10~30min,减压浓缩,冻干,得水提物干粉。
2.一种富硒香菇提取物的应用,其特征在于,将权利要求1所述的制备方法制备的富硒香菇提取物应用于抗癌或辅助治疗作用。
3.如权利要求2所述的应用,其特征在于,所述抗癌或辅助治疗作用对象包括:人前列腺癌(LNCaP),肝癌(HepG2)、肺癌(A549)、宫颈癌(Hela)、白血病(K562)。
4.如权利要求2或3所述的应用,其特征在于,将所述水提物干粉单独或组合到其他食品或药品中,应用于抗癌或辅助治疗作用。
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