CN110974853B - 基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物及应用 - Google Patents
基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物及应用 Download PDFInfo
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Abstract
一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物及具有该组合物的抗癌功能物品及用途,基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,含有罗伊氏乳杆菌A2发酵上清液和姜黄素,以体积份计,罗伊氏乳杆菌A2发酵上清液:姜黄素溶液=300~1:3~1,其中姜黄素溶液的浓度为40~300mM。抗癌功能物品,具有基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物。优选的,上述抗癌功能物品为抗癌药物、抗癌食品或者抗癌保健品,所述抗癌功能物品为制剂、粉末或者片剂。本发明含有罗伊氏乳杆菌A2发酵上清液和姜黄素。该组合物与多种人肿瘤细胞作用后,可以降低肿瘤细胞的活力,具有抗癌作用。
Description
技术领域
本发明涉及生物技术领域,特别是涉及一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物及作为抗癌功能物品包括药物、保健品等及其用途。
背景技术
癌是一种恶性肿瘤。发生于人与动物体组织,器官的细胞无限制增生,导致对附近正常组织的压挤,侵犯和毁坏。增生的癌细胞可由血液和淋巴带至身体其他部位进行增殖与破坏。癌症的治疗中,药物治疗是一个很重要的环节,有效的抗癌药物的使用,可以帮助患者获得更长的生存时间,拥有活下来的希望,目前最为常见的抗癌药物有化疗药物、中药、生物制药、靶向药物等。各种药物的作用方式和效果也有不同差异。
抗癌功能产品的研究,一直是一个重要研究方向。针对现有技术不足,提供一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物及其作为抗癌功能物品包括但不限于药物、保健品等及其用途以克服现有技术不足甚为必要。
发明内容
本发明的目的之一在于避免现有技术的不足之处而提供一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,具有抗癌功能。
本发明的目的通过以下技术措施实现。
提供一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,含有罗伊氏乳杆菌A2发酵上清液和姜黄素。
优选的,上述的基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,以体积份计,罗伊氏乳杆菌A2发酵上清液:姜黄素溶液=300~1:3~1,其中姜黄素溶液的浓度为40~300mM。
优选的,上述的罗伊氏乳杆菌A2发酵上清液与姜黄素溶液的体积比为300:1。
优选的,上述的罗伊氏乳杆菌A2为罗伊氏乳杆菌A2菌株,所述罗伊氏乳杆菌A2菌株于2019年9月02日保藏于广东省微生物菌种保藏中心(GDMCC),保藏编号为GDMCC NO:60758。地址:广东省广州市先烈中路100号大院59号楼5楼,邮编:510070。
优选的,罗伊氏乳酸菌发酵上清液的制备过程具体是:采用MRS固体培养基活化菌种于37℃培养箱培养48h;之后将活化好的菌株取2环接种到MRS肉汤培养基中,再于37℃发酵48h,调整发酵后菌液的OD至1.8;然后在4℃条件下,以转速6000r/min离心15min,留取上清,待用。
本发明提供了一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,含有罗伊氏乳杆菌A2发酵上清液和姜黄素。该组合物与多种人肿瘤细胞作用后,可以降低肿瘤细胞的活力,具有抗癌作用。
本发明的另一目的在于提供一种抗癌功能物品,可以降低肿瘤细胞的活力,具有抗癌作用。
本发明的上述目的通过如下技术手段实现:
提供一种抗癌功能物品,具有基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物。
优选的,上述抗癌功能物品为抗癌药物、抗癌食品或者抗癌保健品,所述抗癌功能物品为制剂、粉末或者片剂。
另一优选的,上述抗癌功能物品能抑制癌细胞的增值功能。
优选的,能抑制癌细胞的增值功能的抗癌功能物品为饮品;
饮品的制备方法包括以下步骤:按照体积份数称取罗伊氏乳杆菌A2发酵上清液和姜黄素溶液,所称取的罗伊氏乳杆菌A2发酵上清液和姜黄素溶液的总体积为V份,V为正数,将罗伊氏乳杆菌A2发酵上清液和姜黄素溶液溶解于80~100V份水中,得到具有抗癌功能的饮品。
本发明的抗癌功能物品,具有基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,该物品,可以降低肿瘤细胞的活力,具有抗癌作用。
本发明同时提供基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物作为抗癌功能物品方面的用途,所述抗癌功能物品为抗癌药物、抗癌食品或者抗癌保健品,所述抗癌功能物品为制剂、粉末或者片剂。
附图说明
利用附图对本发明作进一步的说明,但附图中的内容不构成对本发明的任何限制。
图1是本发明一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物对肠癌的抑制作用实验结果图。
图2是本发明一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物对肺癌的抑制作用实验结果图。
图3是本发明一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物对乳腺癌的抑制作用实验结果图。
具体实施方式
结合以下实施例对本发明作进一步说明。
实施例1。
一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,含有罗伊氏乳杆菌A2发酵上清液和姜黄素。
具体的,以体积份计,罗伊氏乳杆菌A2发酵上清液:姜黄素溶液=300~1:3~1,其中姜黄素溶液的浓度为40~300mM。
罗伊氏乳杆菌A2为罗伊氏乳杆菌A2菌株,罗伊氏乳杆菌A2菌株于2019年9月02日保藏于广东省微生物菌种保藏中心(GDMCC),保藏编号为GDMCCNO:60758。地址:广东省广州市先烈中路100号大院59号楼5楼,邮编:510070。
其中,罗伊氏乳酸菌发酵上清液的制备过程具体是:采用MRS固体培养基活化菌种于37℃培养箱培养48h;之后将活化好的菌株取2环接种到MRS肉汤培养基中,再于37℃发酵48h,调整发酵后菌液的OD至1.8;然后在4℃条件下,以转速6000r/min离心15min,留取上清,待用。
本实施例的基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,含有罗伊氏乳杆菌A2发酵上清液和姜黄素。该组合物与多种人肿瘤细胞作用后,可以降低肿瘤细胞的活力,具有抗癌作用。
实施例2。
一种基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,其它结构与实施例1相同,不同之处在于:罗伊氏乳杆菌A2发酵上清液与姜黄素溶液的体积比为300:1,姜黄素溶液的浓度为40~300mM。
实验发现,罗伊氏乳杆菌A2发酵上清液与姜黄素溶液的体积比为300:1时,抗癌效果最佳。
实施例3。
提供一种抗癌功能物品,具有如实施例1或2任意一项所述的基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物。
抗癌功能物品可以为抗癌药物、抗癌食品或者抗癌保健品,抗癌功能物品为制剂、粉末或者片剂。
该抗癌功能物品,含有基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,该物品,可以降低肿瘤细胞的活力,具有抗癌作用。
实施例4。
抗癌功能物品能抑制癌细胞的增值功能,含有基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,能抑制癌细胞的增值功能的抗癌功能物品为饮品。
饮品的制备方法包括以下步骤:按照体积份数称取罗伊氏乳杆菌A2发酵上清液和姜黄素溶液,所称取的罗伊氏乳杆菌A2发酵上清液和姜黄素溶液的总体积为V份,V为正数,将罗伊氏乳杆菌A2发酵上清液和姜黄素溶液溶解于80~100V份水中,得到具有抗癌功能的饮品。
该能抑制癌细胞的增值功能的抗癌功能物品,可以降低肿瘤细胞的活力,具有抗癌作用。
实施例5。
基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物作为抗癌功能物品方面的用途,抗癌功能物品含有实施例1或2任意一项所述的基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物,抗癌功能物品为实施例3或4任意一项所述的为抗癌药物、抗癌食品或者抗癌保健品,抗癌功能物品为制剂、粉末或者片剂。
基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物可以作为抗癌功能物品使用,能够起到良好的抗癌作用。
实施例6。
下面结合具体实验过程对本发明作进一步说明。
第一部分,罗伊氏乳杆菌A2的分离培养及酵发酵液的制备
1.培养基配置
MRS固体培养基:向蛋白胨10.0g、牛肉粉8.0g、酵母粉4.0g、葡萄糖20.0g、磷酸氢二钾2.0g、乙酸钠5.0g、柠檬酸二铵2g、硫酸镁0.2g、硫酸锰0.05g、吐温80 1g、琼脂15g整体中加入1L水,搅拌均匀,并于121℃高压灭菌15min,倒置平板里等其凝固备用;
MRS肉汤培养基:向蛋白胨10.0g、牛肉粉8.0g、酵母粉4.0g、葡萄糖20.0g、磷酸氢二钾2.0g,乙酸钠5.0g,柠檬酸二铵2g、硫酸镁0.2g、硫酸锰0.05g、吐温80 1g整体中加入1L水,搅拌均匀,并于121℃高压灭菌15min;
2.罗伊氏乳杆菌A2菌株的分离培养
2.1划线在近火焰处,左手拿皿底,右手拿接种环,按无菌操作对平板进行划操作。
2.2用接种环以无菌操作挑取菌液一环,先在平板培养基的一边作第一次平行划线3-4条,再转动培养皿约70°角,并将接种环上剩余物烧掉,待冷却后通过第一次划线部分做第二次平行划线,再用同法通过第二次平行划线部分做第三次平行划线和通过第三次平行划线部分作第四次平行划线。划线完毕后,盖上皿盖,倒置于放在40℃恒温培养箱中培养48小时。
3.罗伊氏乳杆菌A2培养方法及发酵液的制备
采用MRS固体培养基活化菌种,37℃培养箱培养48h;之后将活化好的菌株取2环接种到MRS肉汤培养基中,37℃发酵48h。调整发酵后菌液的OD至1.8;在4℃下,以6000r/min离心15min,留取上清,待用。
第二部分,姜黄素和罗伊氏乳杆菌A2发酵液对人多种肿瘤细胞影响的研究实验
1、溶液的配备:
(1)TMAO溶液:称量75.11mg无水氧化三甲胺粉末,加入10mL PBS,用超声充分混匀,得到浓度为100mM的TMAO溶液,将其分装至10个1.5mL的EP管中,每管1mL,放置于-20℃冰箱。
(2)姜黄素溶液:称量552.57mg姜黄素粉末,加入5mL DMSO混匀,得到浓度为300mM的姜黄素溶液,分装后放于-20℃冰箱保存。
2、CCK8法检测人多种肿瘤细胞活力的方法:
肝癌细胞Hepg2、乳腺癌细胞MCF生长于含10%胎牛血清的DMEM高糖培养基中,肠癌细胞HCT116生长于含10%胎牛血清的1640培养基中,在条件为37℃,5%CO2和完全饱和湿度的细胞培养箱中进行培养,隔一天换液以维持良好生长状态。把对数生长期细胞以5×103个细胞/孔接种于96孔板中,每孔接种100μl细胞悬液,每个时间点设置5个复孔。往边缘孔加入200μl无菌PBS缓冲液防止培养基蒸发。用或者不用含培养基(浓度40mM)或者姜黄素(浓度40mM,体积1份)加罗伊氏乳酸菌A2上清(OD值为1.8,体积3-300ul),培养基100ul。
在培养箱内培养至相应时间点后,抽掉培养基,用PBS洗三遍,加入新的无药物培养基100ul,接着每孔加入10μl cck8增强型溶液,37℃孵育2h。使用酶标仪下测定各孔样品在450nm波长下的吸光度值。实验结果以均值±标准差表示,以时间为横坐标,细胞存活率百分比值为纵坐标做折线图。
注:计算公式细胞存活率=[(As-Ab)/(Ac-Ab)]×100%;
As:实验孔(含有细胞的培养基、CCK-8、待测药物)的吸光度;
Ac:对照孔(含有细胞的培养基、CCK-8、没有待测药物)的吸光度;Ab:空白孔(不含细胞和待测药物的培养基、CCK-8)的吸光度。
3.实验结果
实验结果显示,本发明的基于姜黄素和罗伊氏乳酸菌发酵上清液的抗癌组合物对癌细胞具有良好的抑制作用,抗癌效果明显。本发明的抗癌组合物在罗伊氏乳杆菌A2上清液与姜黄素溶液的比例在100:1至300:1范围时,本发明的抗癌组合物对癌细胞的抑制作用优于单独的姜黄素溶液对癌细胞的抑制作用。
实验还发现,姜黄素溶液和罗伊氏乳杆菌A2上清液比例为1:300时,本发明的抗癌组合物能对肿瘤细胞活力降低达到一个最大值,如图1至图3所示。
最后应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
Claims (6)
1.一种基于姜黄素和罗伊氏乳酸菌发酵上清液的组合物在制备抗癌药物中的用途,其特征在于:所述组合物含有罗伊氏乳杆菌A2发酵上清液和姜黄素;
所述罗伊氏乳杆菌A2为罗伊氏乳杆菌A2菌株,所述罗伊氏乳杆菌A2菌株于2019年9月02日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC NO:60758。
2.根据权利要求1所述的用途,其特征在于:所述基于姜黄素和罗伊氏乳酸菌发酵上清液的组合物,以体积份计,罗伊氏乳杆菌A2发酵上清液:姜黄素溶液=300~1:3~1,其中姜黄素溶液的浓度为40~300mM。
3.根据权利要求2所述的用途,其特征在于:罗伊氏乳杆菌A2发酵上清液与姜黄素溶液的体积比为300:1。
4.根据权利要求1至3任意一项所述的用途,其特征在于:罗伊氏乳酸菌发酵上清液的制备过程具体是:采用MRS固体培养基活化菌种于37℃培养箱培养48h;之后将活化好的菌株取2环接种到MRS肉汤培养基中,再于37℃发酵48h,调整发酵后菌液的OD至1.8;然后在4℃条件下,以转速6000r/min离心15min,留取上清,待用。
5.根据权利要求4所述的用途,其特征在于:组合物制成粉末或者片剂。
6.根据权利要求5所述的用途,其特征在于:组合物抑制癌细胞的增殖功能。
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