CN106361838A - 一种含赤芍和黄芪的抗肿瘤组合物、制备方法和应用 - Google Patents
一种含赤芍和黄芪的抗肿瘤组合物、制备方法和应用 Download PDFInfo
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Abstract
本发明涉及一种含赤芍和黄芪的抗肿瘤组合物、制备方法和应用,属于中药技术领域。组合物包括有按照重量百分比计的如下组分:赤芍提取物80~94%、黄芪多糖1~18%、竹节参皂苷0.1~5%。本发明提供了一种赤芍提取物和黄芪多糖的组合物,通过黄芪多糖的增效作用可以显著提高赤芍提取物对于抑制原癌基因ras通路过度激活的效果,通过竹节参皂苷用于减轻组合物毒性。该组合物对于Ras基因具有明显的抑制调节作用。
Description
技术领域
本发明涉及一种含赤芍和黄芪的抗肿瘤组合物、制备方法和应用,属于中药技术领域。
背景技术
赤芍 Paeoniae Rubra Radix(PRR)为毛茛科(Ranunculaceae)植物芍药 Paeonia lactiflora Pall. 或川赤芍 Paeonia veitchii Lynch 的干燥根。其味苦,性微寒,归肝经,具有清热凉血、散瘀镇痛之功效[1]。 现代研究表明,赤芍包含萜类及其苷、黄酮类及其苷、挥发油类等多种化学成分,具有保肝、抗肿瘤(吴杨峥等)、神经保护、心脏保护、抗血栓、抗氧化、抗内毒素等多种药理作用[2]。
于晓红等通过观察赤芍总苷对S180荷瘤鼠CD4、CD8分子的表达,探讨了赤芍总苷的抗肿瘤效应的免疫学机理[3]。吴明媛等发现赤芍总苷能够明显增强荷瘤机体NK细胞的杀伤活性,激活巨噬细胞并提高TNF-α的分泌,拮抗环磷酰胺引起的免疫抑制状态,改善IL-2/IL-4的平衡状态,纠正Thl/Th2漂移,显著提高荷瘤机体IL-2的产生,使其接近正常水平,作为激动剂激发机体免疫应答[4]。孙国平等[5]报道芍药有效成分之一丹皮酚能够提高荷瘤小鼠IL-2和TNF-α的表达,从而对肿瘤细胞产生杀伤和抑制作用。许惠玉等[6]发现赤芍总苷能够明显降低Bcl-2蛋白的表达和c-myc基因mRNA的表达,同时上调p16蛋白的表达水平,并且不同浓度赤芍总苷引起细胞内钙离子的浓度升高,线粒体膜电位下降,K562细胞凋亡明显。
然而,赤芍提取物对于抗肿瘤的效果仍然有进一步提高的必要性。
发明内容
本发明提供了一种含有赤芍的中药组合物,通过黄芪多糖的加入可以显著地提高其对于抑制原癌基因ras通路过度激活的效果。
技术方案是:
一种含赤芍和黄芪的抗肿瘤组合物,包括有按照重量百分比计的如下组分:赤芍提取物80~94%、黄芪多糖1~18%、竹节参皂苷0.1~5%。
所述的竹节参皂苷是指竹节参皂苷III、竹节参皂苷IV或者竹节参皂苷V。
所述的赤芍提取物,它是由赤芍经过水煎之后,将煎液由石油醚、乙酸乙酯或者正丁醇萃取、减压蒸馏、干燥后而得到。
所述的黄芪多糖是黄芪的干燥根经提取、浓缩、纯化而成的水溶性杂多糖。
所述的组合物的制备方法,是将赤芍提取物、黄芪多糖和竹节参皂苷混合均匀后而得到。
上述的抗肿瘤组合物在制备抗肿瘤药物中的应用。
所述的抗肿瘤药物,是指抑制原癌基因ras通路过度激活的抗肿瘤药物。
有益效果
本发明提供了一种赤芍提取物和黄芪多糖的组合物,通过黄芪多糖的增效作用可以显著提高赤芍提取物对于抑制原癌基因ras通路过度激活的效果,通过竹节参皂苷用于减轻组合物毒性。该组合物对于Ras基因具有明显的抑制调节作用。
具体实施方式
1、仪器、试剂及实验材料
1.1 仪器
GR60DR型高温灭菌锅(致微仪器有限公司),B2生物安全柜(力康生物医疗科技控股有限公司),底温高速离心机,SY-2OOO型旋转蒸发器(上海亚荣生化仪器有限公司),CHZQF(60A)型气浴恒温摇床,DZF-6020MBE型真空干燥箱,HHS型恒温水浴锅(余姚市检测仪表厂),分析天平(北京赛多利斯仪器系统有限公),倒置生物显微镜(重庆光电仪器总公司)。
1.2 试剂
石油醚(沸点:30℃-60℃,分析纯,天津市百世化工有限公司),乙酸乙酯(分析纯,天津市百世化工有限公司),正丁醇(分析纯,天津市百世化工有限公司),无水乙醇(分析纯,天津市百世化工有限公司),二甲基亚砜(分析纯,上海中泰化学试剂有限公司)。
1.3 实验材料
药材:市售赤芍,购于兰州市黄河药材市场。
黄芪多糖:市售,购于上海远慕生物科技有限公司。
线虫株:MTZ124,基因型:let-60(nlo46sd,g f)IV, 购于美国线虫遗传中心CGC(Caenorhabditis Genetics Center)。
大肠杆菌:E.coli OP50,尿嘧啶渗漏突变株,作为秀丽隐杆线虫的食物,购于CGC。
1.4 溶液的配置
NGM培养基:100ml纯净水加入0.3031g氯化钠,1.7033g磷酸二氢钾,0.2309g磷酸氢二钾,0.27185g蛋白胨,1.7155g琼脂,使其全部溶解。(灭菌后加0.2ml氯化钙,0.2ml硫酸镁,0.1ml胆固醇。)
S基础液:100ml纯净水加0.585g氯化钠,0.1g磷酸氢二钾,0.6g磷酸二氢钾,混合均匀灭菌。S液:100ml S基础液加入1ml柠檬酸,1mlTrace,100μl胆固醇,300μl硫酸镁,300μl氯化钙,混合均匀。(现配现用)
LB培养基:100ml纯净水加入1.0g蛋白胨,0.5g酵母粉,1.0g氯化钠,混合均匀灭菌(再加1.7155g琼脂,灭菌,得LB固体培养基)。
M9缓冲液:100ml纯净水加0.49725g氯化钠,0.5964g磷酸氢二钠,0.2992g磷酸二氢钾,0.012g硫酸镁,混合均匀,灭菌。
裂解液:1M氢氧化钠:2.4%次氯酸钠按1 :1的比例混合。
实施例1 赤芍提取物的制备
取赤芍1Kg,加入10Kg水中浸泡1h,再煎煮3次,每次1h,每次煎煮滤除滤液并加入10Kg水,合并煎液后,依次用煎液重量20%的石油醚、乙酸乙酯、正丁醇萃取,将三步得到的萃取液分别减压蒸馏除溶剂后,再干燥后得到提取物。
实施例2 抗肿瘤组合物的制备
按照下表中的重量配制组合物,混合均匀后,得到组合物。
。
对照例1
与实施例2的区别是:黄芪多糖未加入,其重量是由竹节参皂苷替代,混合均匀后,得到组合物。
对照例2
与实施例2的区别是:竹节参皂苷未加入,其重量是由黄芪多糖,混合均匀后,得到组合物。
对照例3
与实施例2的区别是:赤芍提取物未加入,混合均匀后,得到组合物。
实施例3药效试验
大肠杆菌OP50培养
将从线虫遗传中心购置的在固体培养基上培养的OP50,用接种环接种到LB液体培养基中,放在摇床上,培养条件200rpm,20℃,培养24h,一部分用于涂菌,每板加400μl菌液,用涂菌棒涂布均匀,放置恒温培养箱中培养24h,用于线虫转板;另一部分离心(10000rpm,10min)得到固体菌,将其称重,然后稀释成110mg/ml的菌液,置于4℃冰箱保存。
线虫培养
将冻存于-80℃冰箱中的秀丽隐杆线虫MT2124转接到涂有OP50的固体NGM培养基上,在20℃下培养成虫进行同步化。将同步化线虫孵化成L1期小线虫用于药物活性实验的生物模型和急性毒性实验。
线虫同步化
将培养了2-3d的固体板线虫用M9液冲洗至10ml离心管中,使线虫沉降,吸出上清液,先后加入3ml 1M氢氧化钠和3ml 2.4%次氯酸,在涡旋器上涡旋6min,快速吸入1.5ml的离心管中,离心(4min 3000rpm),快速吸出裂解液,再用M9液冲洗两次,最后加约1ml M9液,放置恒温箱中培养24h后。同步化液体线虫时,吸出S液时用M9液清洗一次,再加裂解液,后面步骤跟固体板线虫相同。
对线虫突变体的抑制效果
将实施例2、对照例1~3的组合物用S基础液稀释成赤芍提取物浓度为5000μg/ml的溶液;再用S基础液将110mg/ml的OP50稀释成11mg/ml的菌液;同时把培养2d的同步化线虫用S基础液稀释成60-80条每20μl的线虫液。在96孔板的药物组的每一药孔中加160μl均匀药液,20μl OP50,20μl线虫,在空白组的每孔中加160μl S基础液,20μl OP50,20μl线虫,然后将其混合均匀,放置恒温振荡器中培养,不断观察,待线虫培养成成虫后,观察统计药物组与空白组中野生型的比例(野生型比例=野生型/线虫总数)[7],然后将药物组与空白组进行比较,判断药物对线虫突变体是否有抑制作用。
线虫中的野生型和突变体的判定
由于成虫野生型和突变体线虫真假阴门表型很容易在倒置显微镜下观察和区别,阴门发育从L1期开始,成虫期时发育完全,且便于观察。
实施例和对照例的组合物对秀丽隐杆线虫ras过度激活突变体MT2124多阴门表型
的影响
将以上稀释成后得到的药液作用于L1期线虫,将所有组线虫培养至成虫后,在倒置显微镜下观察并统计其野生型比例。
野生型比例试验结果如下:
*相对于对照例1具有显著性p<0.01,▲相对于对照例2具有显著性P<0.01,△相对于空白S基础液没有显著性p<0.01。
通过实施例2与对照例2相比可以看出,通过在实施例中加入了竹节参皂苷IV,使赤芍提取物对于线虫的野生型比例得到了显著的提高,说明竹节参皂苷IV能够提高赤芍提取物的作用;通过对照例3和空白S基础液的对照可以看出,竹节参皂苷IV在一定程度上抑制ras过度激活但效果较差 。同时,通过实施例2与对照例1相比也可以看出,通过黄芪多糖的加入也同时能够与赤芍提取物产生协同效果,提高抑制ras基因的作用。
急性毒性实验方法
母液:称取实施例和对照例中的组合物,配制成含有赤芍提取物浓度在100mg/ml的乙醇溶液。
做药板:在96孔板上加入药液和线虫,每个药液供试品设三个平行,将溶解药物的试剂(无水乙醇)做为阴性对照组,并设置空白对照组。
计算死亡率:将做好的药板放入恒温培养箱(20℃)培养,24h后用倒置显微镜观察线虫死亡情况,计算死亡比例。
野生型比例试验结果如下:
*与对照例1组相比具有显著性p<0.01,
从表中可以看出,实施例相对于对照例1来说,通过加入了黄芪多糖可以显著地降低了赤芍提取物的毒性,使线虫的死亡率降低。
[1]中国药典[S]. 一部. 2010.
[2]陆小华, 马骁, 王建, 朱云, 周宗元, 陈哲, 赵艳玲, 王伽伯. 赤芍的化学成分和药理作用研究进展[J]. 中草药, 2015, v.46; No.54304: 595-602.
[3]于晓红, 于洋, 华东. 赤芍总苷对荷瘤小鼠抗肿瘤作用的免疫机理研究[J]. 江西中医学院学报,2005, 01:61-62.
[4]吴明媛. 赤芍总甙抗肿瘤作用的免疫机理研究[D]. 黑龙江中医药大学, 2002.
[5]Sun G P, Wang H, Xu S P.ET al. Anti-tumor effects of paeonol in a HepA-hepatoma bearing mouse model via induction of tumor cellapoptosis andstimulation of IL-2 and TN F-alpha production. 〔J〕.Eur J Pharmacologist,2008, 584(2-3): 246-252.
[6]许惠玉. 赤芍总苷抗S180和K562肿瘤细胞作用机制的实验研究[D]. 北京中医药大学, 2008.
[7] 丁晓霞. 以秀丽隐杆线虫作为模式生物对雄黄微生物浸出液活性和毒性的研究[D]. 2009.。
Claims (7)
1.一种含赤芍和黄芪的抗肿瘤组合物,其特征在于,包括有按照重量百分比计的如下组分:赤芍提取物80~94%、黄芪多糖1~18%、竹节参皂苷0.1~5%。
2.根据权利要求1所述的含赤芍和黄芪的抗肿瘤组合物,其特征在于,所述的竹节参皂苷是指竹节参皂苷III、竹节参皂苷IV或者竹节参皂苷V。
3.根据权利要求1所述的含赤芍和黄芪的抗肿瘤组合物,其特征在于,所述的赤芍提取物,它是由赤芍经过水煎之后,将煎液由石油醚、乙酸乙酯或者正丁醇萃取、减压蒸馏、干燥后而得到。
4.根据权利要求1所述的含赤芍和黄芪的抗肿瘤组合物,其特征在于,所述的黄芪多糖是黄芪的干燥根经提取、浓缩、纯化而成的水溶性杂多糖。
5.权利要求1~4任一项所述的含赤芍和黄芪的抗肿瘤组合物的制备方法,其特征在于,是将赤芍提取物、黄芪多糖和竹节参皂苷混合均匀后而得到。
6.权利要求1~4任一项所述的含赤芍和黄芪的抗肿瘤组合物在制备抗肿瘤药物中的应用。
7.根据权利要求6所述的应用,其特征在于,所述的抗肿瘤药物,是指抑制原癌基因ras通路过度激活的抗肿瘤药物。
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