CN114558053B - 一种具有治疗胃癌作用的废弃果皮提取物组合物 - Google Patents
一种具有治疗胃癌作用的废弃果皮提取物组合物 Download PDFInfo
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Abstract
本发明涉及一种具有治疗胃癌作用的废弃果皮提取物组合物,该组合物由番石榴皮提取物、释迦皮提取物和百香果皮提取物按照4:2:3的比例组成。该组合物均来自于废弃果皮,来源丰富成本低,提取方法简单易实现工业化生产,可充分利用废弃资源,既减少环境污染,又能提高水果的经济附加值。该组合物各组分间发挥明显的协同增效作用,能显著抑制胃癌细胞的增殖、侵袭和迁移,可用于制备治疗胃癌的药物,既能创造更高的经济效益,又具有良好的社会效益。
Description
技术领域
本发明涉及一种具有治疗胃癌作用的废弃果皮提取物组合物,更具体地涉及一种由由番石榴皮提取物、释迦皮提取物和百香果皮提取物组成的具有治疗胃癌作用的废弃果皮提取物组合物。
背景技术
由于人口老龄化和越来越多的人采用易致癌的生活方式行为,导致全球癌症负担持续增加。在各种癌症中,胃癌(gastric cancer,GC)是最常见的恶性肿瘤之一,是世界第四大常见癌症,是导致癌症死亡的第二大原因,成为重大的公共卫生问题。胃癌是一种侵袭性疾病,其预后较差。由于无特异性症状,导致早期诊断非常困难。根据2020年发布的一份报告,全球每年诊断出的胃癌病例超过100万例。约有一半的胃癌患者在手术治疗后有复发性疾病,但这些患者的预后仍很差,局部疾病的5年生存率估计为25%-35%。因此,发现新的药物抑制胃癌的侵袭和转移,延长胃癌患者的生存时间,提高患者的生存质量,具有重要意义。
番石榴是桃金娘科番石榴属乔木,原产南美洲。中国华南各地栽培,鲜果可供食用,叶和果也可入药,具有收敛止泻,消炎止血的功效。春、夏采叶,秋季采果,晒干。叶含挥发油及鞣质等,有止痢、止血、健胃等功效;叶经煮沸去掉鞣质,晒干作茶叶用,味甘,有清热作用。食用的鲜果果皮通常丢弃,既污染环境,又造成资源的浪费。
释迦,是番荔枝科、番荔枝属落叶小乔木,原产热带美洲,现全球热带地区均有栽培。其鲜果可食用,营养丰富,含有大量蛋白质、糖类、矿物质、维生素、氨基酸等营养物质,可提高机体免疫力,促进身体的生长发育;还含有大量的膳食纤维,可以促进肠胃的蠕动,清除宿便,缓解便秘的症状;同时,释迦果的热量非常高,因此也可以及时的补充体力;此外,释迦果还具有美容养颜、强筋健骨的功效。释迦果皮在食用时被丢弃,既污染环境,又造成资源的浪费。
百香果,又名鸡蛋果,是西番莲科、西番莲属草质藤本植物。其根、茎、叶均可入药,有兴奋、强壮、除风清热提神醒脑、美容养颜、滋补健身、防治胃、肠道炎症、镇静止痛、提高人体免疫力等功能,还有减压降脂和延缓衰老等功效。果实内有黄色果汁和黑色种子,含有蛋白质、脂肪、糖类和多种维生素及氨基酸等,其中可溶性固形物15-16%,总酸量3.8-4.0%,享有“饮料味精”和“果汁之王”的美誉。百香果可鲜食或加工成果汁、果露、果酱、果冻、口含片等。其果皮厚而坚硬,通常丢弃,也有用作饲料和提取果胶。
为了充分利用废弃果皮资源,提高水果的经济附加值,本发明人经过大量的活性筛选实验和配方比例的研究,最后获得了本发明最佳比例的废弃果皮提取物组合物,并证明了其在抑制胃癌细胞增殖、侵袭和迁移中的积极作用,由此完成了本发明。本发明提供了一种废弃果皮提取物组合物,该组合物具有显著的体外抗胃癌活性,可用于制备治疗胃癌的药物。
发明内容
针对上述情况,为了克服现有技术的不足,本发明的目的是为了充分利用废弃果皮资源,同时提供一种新的治疗胃癌的药物。
为了达到上述目的,本发明提供了一种具有治疗胃癌作用的废弃果皮提取物组合物。
本发明解决的技术方案是,提供一种具有治疗胃癌作用的废弃果皮提取物组合物,该组合物是由番石榴皮提取物、释迦皮提取物和百香果皮提取物按照4:2:3的比例组成;
其中番石榴皮提取物的制备方法是:
取番石榴皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入6倍量80%乙醇加热连续回流提取2次,每次2h,过滤,合并滤液,滤液置于旋转蒸发仪中回收乙醇至无醇味,浓缩,干燥,即得番石榴皮提取物;
其中释迦皮提取物的制备方法是:
取释迦皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入10倍量蒸馏水加热煮沸2次,每次1h,过滤,合并滤液,滤液加热浓缩,干燥,即得释迦皮提取物;
其中百香果皮提取物的制备方法是:
取百香果皮,晒干,粉碎,过40目筛,置于三角瓶中,加入5倍量的蒸馏水,加热至60℃温浸3h,然后加入0.5%的果胶酶,用盐酸调节pH至3.0后,置于50℃恒温振荡器中酶解30min,然后转入超声提取器中,设置超声频率30kHz,温度为60℃,超声提取50min,过滤,滤液加热浓缩,干燥,即得百香果皮提取物。
按4:2:3的比例选取番石榴皮提取物、释迦皮提取物和百香果皮提取物,混合均匀,即得一种具有治疗胃癌作用的废弃果皮提取物组合物。
本发明相比现有技术具有以下优点:
1.本发明提取物组合物均来自于废弃果皮,来源丰富成本低,提取方法简单易实现工业化生产,可充分利用废弃资源,既减少环境污染,又能提高水果的经济附加值,创造更高的经济效益。
2.本发明人独创地将番石榴皮提取物、释迦皮提取物和百香果皮提取物进行组合,并通过大量实验进行活性筛选,发现了组合物抑制胃癌细胞增殖、侵袭和迁移的作用,并确定了各组分之间的用量配比,使其发挥明显的协同增效作用,可用于制备治疗胃癌的药物。
附图说明
图1为废弃果皮提取物组合物对胃癌HGC-27细胞侵袭能力的影响。
图2为废弃果皮提取物组合物对胃癌HGC-27细胞迁移能力的影响。
具体实施方式
下面结合具体实施例对本发明进行详细说明。
实施例1废弃果皮提取物组合物的制备
取番石榴皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入6倍量80%乙醇加热连续回流提取2次,每次2h,过滤,合并滤液,滤液置于旋转蒸发仪中回收乙醇至无醇味,浓缩,干燥,即得番石榴皮提取物;
取释迦皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入10倍量蒸馏水加热煮沸2次,每次1h,过滤,合并滤液,滤液加热浓缩,干燥,即得释迦皮提取物;
取百香果皮,晒干,粉碎,过40目筛,置于三角瓶中,加入5倍量的蒸馏水,加热至60℃温浸3h,然后加入0.5%的果胶酶,用盐酸调节pH至3.0后,置于50℃恒温振荡器中酶解30min,然后转入超声提取器中,设置超声频率30kHz,温度为60℃,超声提取50min,过滤,滤液加热浓缩,干燥,即得百香果皮提取物。
按4:2:3的比例选取番石榴皮提取物、释迦皮提取物和百香果皮提取物,混合均匀,即得一种具有治疗胃癌作用的废弃果皮提取物组合物。
以下通过试验例进一步阐述本发明所述的治疗胃癌作用。
试验例1废弃果皮提取物组合物对胃癌HGC-27细胞活力的影响
1.试验材料
按实施例制备的废弃果皮提取物组合物,分别加DMEM培养基溶解,配制成0.125、0.25、0.5、1和2mg/mL的溶液;
青霉素链霉素溶液,胎牛血清,DMEM培养基,PBS缓冲液,噻唑蓝(MTT),二甲基亚砜(DMSO),胃癌HGC-27细胞和人脐静脉内皮细胞(HUVEC)购自碧云天生物技术有限公司。
2.试验方法
采用MTT法检测细胞活力。将HGC-27细胞和HUVEC细胞(4×103细胞/孔)接种于96孔板,置于含5%CO2培养箱中37℃培养24h后,每孔分别加入0.125、0.25、0.5、1和2mg/mL的溶液各100μL,培养24h后每孔加入20μL 0.5%MTT溶液,继续培养4h后终止培养,小心吸去孔内培养液,每孔加入150μL DMSO,置摇床上低速振荡10min,使结晶物充分溶解,用全波长酶标仪测定490nm处的光密度(OD)。以DMEM培养基作对照,每孔设置5个复孔。以DMEM培养基作对照,计算相对细胞活力(%)。
3.试验结果
废弃果皮提取物组合物对HUVEC和HGC-27细胞活力的影响见表1。从表1可以看出,随着废弃果皮提取物组合物浓度的增加,HUVEC和HGC-27细胞的活力均逐渐下降。提取物组合物浓度从0.25mg/mL起,HGC-27细胞的活力就降至70%以下,至2mg/mL时,细胞活力为21.33±8.87%,提示废弃果皮提取物组合物对HGC-27细胞具有明显的细胞毒作用。而在HUVEC细胞,提取物组合物浓度为0.5mg/mL时,细胞活力仍为在80%以上,至1mg/mL时细胞活力降至70%以下,对HUVEC细胞产生毒性。从实验结果看,废弃果皮提取物组合物浓度在0.5mg/mL时,对HGC-27细胞具有明显毒性而对HUVEC细胞无明显毒性,因此,废弃果皮提取物组合物浓度定在0.5mg/mL,用于考察其抑制胃癌HGC-27细胞侵袭和迁移的作用。
表1废弃果皮提取物组合物对HUVEC和HGC-27细胞活力的影响(均值±标准差)
4.结论
废弃果皮提取物组合物可抑制HGC-27细胞活力,并且具有剂量依赖关系。
试验例2废弃果皮提取物组合物对胃癌HGC-27细胞侵袭能力的影响
1.试验材料
按实施例制备的废弃果皮提取物组合物,分别加DMEM培养基溶解,配制成0.5mg/mL的溶液,即为本发明提取物组合物(组合物1);
按组方比例制成番石榴皮提取物和释迦皮提取物组合物(组合物2,百香果皮提取物用等量糖粉代替);按组方比例制成番石榴皮提取物和百香果皮提取物组合物(组合物3,释迦皮提取物用等量糖粉代替);按组方比例制成释迦皮提取物和百香果皮提取物组合物(组合物4,番石榴皮提取物用等量糖粉代替)。
青霉素链霉素溶液,胎牛血清,DMEM培养基,PBS缓冲液,多聚甲醛,结晶紫,胃癌HGC-27细胞购自碧云天生物技术有限公司;
Matrigel胶购自美国BD公司。
2.试验方法
采用Transwell法检测细胞侵袭能力。将100μL的Matrigel加入12孔Transwell板(8μm,Corning,NY,美国),置于含5%CO2的培养箱中37℃孵育6h。将HGC-27细胞(2×105细胞/孔)接种于上室,分别加入0.5mg/mL的组合物1、组合物2、组合物3和组合物4各100μL,下室加入DMEM完全培养基,置于含5%CO2的培养箱中37℃培养24h。拭去未侵入的细胞,用4%多聚甲醛固定细胞20min,PBS洗涤3次,然后加入0.5%结晶紫染色20min,PBS洗去未与细胞结合的结晶紫,并过夜干燥。使用倒置显微镜拍摄图像,并用33%乙酸溶解形成的结晶紫罗兰,用全波长酶标仪在570nm波长下测定吸光度。以吸光度代表细胞侵入底室的量,以DMEM培养基作对照,计算相对于对照组的细胞侵袭率。
3.试验结果
废弃果皮提取物组合物对胃癌HGC-27细胞侵袭能力的影响见图1。如图所示,与对照组相比,组合物1和组合物2的细胞侵袭率显著下降(P<0.01),说明组合物1和组合物2具有抑制胃癌HGC-27细胞侵袭能力的作用;与组合物1比较,组合物2、组合物3、组合物4的细胞侵袭率显著升高(P<0.05或P<0.01),说明组合物1(本发明废弃果皮提取物组合物)抑制胃癌HGC-27细胞侵袭能力的作用最强,本发明废弃果皮提取物组合物中缺少任何一个组分,都会使其抑制胃癌HGC-27细胞侵袭能力的作用显著下降。
4.结论
本发明废弃果皮提取物组合物能显著抑制胃癌HGC-27细胞侵袭,具有治疗胃癌的作用。
试验例3废弃果皮提取物组合物对胃癌HGC-27细胞迁移能力的影响
1.试验材料
按实施例制备的废弃果皮提取物组合物,分别加DMEM培养基溶解,配制成0.5mg/mL的溶液,即为本发明提取物组合物(组合物1);
按组方比例制成番石榴皮提取物和释迦皮提取物组合物(组合物2,百香果皮提取物用等量糖粉代替);按组方比例制成番石榴皮提取物和百香果皮提取物组合物(组合物3,释迦皮提取物用等量糖粉代替);按组方比例制成释迦皮提取物和百香果皮提取物组合物(组合物4,番石榴皮提取物用等量糖粉代替)。
青霉素链霉素溶液,胎牛血清,DMEM培养基,PBS缓冲液,胃癌HGC-27细胞购自碧云天生物技术有限公司。
2.试验方法
采用划痕实验检测细胞迁移能力。将HGC-27细胞(4×105细胞/孔)接种于6孔板。当细胞接近80-90%融合时,用200μL移液管针尖在单层细胞培养层上造成划痕,倒置显微镜下拍照。用PBS冲洗受损的单层细胞,然后分别加入0.5mg/mL的组合物1、组合物2、组合物3和组合物4各100μL,置于含5%CO2的培养箱中37℃孵育48h,倒置显微镜下拍照。使用Image J软件进行分析,以DMEM培养基作对照。迁移率按下式计算:
3.试验结果
废弃果皮提取物组合物对胃癌HGC-27细胞迁移能力的影响见图2。如图所示,与对照组相比,组合物1和组合物2的细胞迁移率显著下降(P<0.01),说明组合物1和组合物2具有抑制胃癌HGC-27细迁移能力的作用;与组合物1比较,组合物2、组合物3、组合物4的细胞迁移率显著升高(P<0.01),说明组合物1(本发明废弃果皮提取物组合物)抑制胃癌HGC-27细胞迁移能力的作用最强,本发明废弃果皮提取物组合物中缺少任何一个组分,都会使其抑制胃癌HGC-27细胞迁移能力的作用显著下降。
4.结论
本发明废弃果皮提取物组合物能显著抑制胃癌HGC-27细胞迁移,具有治疗胃癌的作用。
Claims (1)
1.一种具有治疗胃癌作用的废弃果皮提取物组合物,其特征在于,所述的废弃果皮提取物组合物由番石榴皮提取物、释迦皮提取物和百香果皮提取物按照4:2:3的比例组成;
其中番石榴皮提取物的制备方法是:
取番石榴皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入6倍量80%乙醇加热连续回流提取2次,每次2h,过滤,合并滤液,滤液置于旋转蒸发仪中回收乙醇至无醇味,浓缩,干燥,即得番石榴皮提取物;
其中释迦皮提取物的制备方法是:
取释迦皮,晒干,粉碎,过40目筛,置于圆底烧瓶中,分别加入10倍量蒸馏水加热煮沸2次,每次1h,过滤,合并滤液,滤液加热浓缩,干燥,即得释迦皮提取物;
其中百香果皮提取物的制备方法是:
取百香果皮,晒干,粉碎,过40目筛,置于三角瓶中,加入5倍量的蒸馏水,加热至60℃温浸3h,然后加入0.5%的果胶酶,用盐酸调节pH至3.0后,置于50℃恒温振荡器中酶解30min,然后转入超声提取器中,设置超声频率30kHz,温度为60℃,超声提取50min,过滤,滤液加热浓缩,干燥,即得百香果皮提取物;
按比例选取番石榴皮提取物、释迦皮提取物和百香果皮提取物,混合均匀,即得一种具有治疗胃癌作用的废弃果皮提取物组合物。
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CN106109584A (zh) * | 2016-06-02 | 2016-11-16 | 中山大学 | 番石榴活性物质的提取工艺及其应用 |
CN108685071A (zh) * | 2017-04-10 | 2018-10-23 | 宋甲祥 | 百香果膏 |
CN113180164A (zh) * | 2021-05-14 | 2021-07-30 | 普洱景谷多上果汁饮品有限公司 | 一种百香果果蔬汁配方及其加工工艺 |
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CN106109584A (zh) * | 2016-06-02 | 2016-11-16 | 中山大学 | 番石榴活性物质的提取工艺及其应用 |
CN108685071A (zh) * | 2017-04-10 | 2018-10-23 | 宋甲祥 | 百香果膏 |
CN113180164A (zh) * | 2021-05-14 | 2021-07-30 | 普洱景谷多上果汁饮品有限公司 | 一种百香果果蔬汁配方及其加工工艺 |
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