苹果类黄酮及其磺化衍生物抗肿瘤作用的研究

发布时间：2018-04-17 12:05

本文选题：苹果类黄酮 + LoVo和MCF-7细胞　；参考：《陕西师范大学》2012年硕士论文

【摘要】：本论文基于合作导师杨兴斌教授的国家自然科学基金项目的资助,测定了粉红女士苹果果肉和果皮的类黄酮总含量,研究了粉红女士苹果类黄酮的体外抗氧化活性。以槲皮素为原料通过磺化反应合成了两种新的槲皮素衍生物槲皮素-5’,8-磺酸钠(QS)和槲皮素-4',7-硫酸钠(QOS)。选用对数生长期的人结肠癌LoVo和乳腺癌MCF-7两种细胞系为实验对象,采用MTT法测定肿瘤细胞的存活率、LDH活力测定法评价黄酮类化合物对肿瘤细胞的毒性、利用Nikon-TS100倒置显微镜观察肿瘤细胞的形态学损伤、Hoechst33258染色法在倒置荧光显微镜下观察肿瘤细胞核的形态变化情况,流式细胞仪分析肿瘤细胞的周期分布和肿瘤细胞中活性氧的产生情况、双变量流式细胞仪检测细胞进入早晚期的凋亡情况,研究了粉红女士苹果类黄酮、芦丁、槲皮素及其衍生物槲皮素-5’,8-磺酸钠(QS)和槲皮素-4',7-硫酸钠(QOS)对人结肠癌LoVo和乳腺癌MCF-7的抗肿瘤活性。研究结果摘要如下： (1)粉红女士苹果果皮、果肉类黄酮的含量分别为24.05±0.14mg/g和10.98±0.25mg/g,类黄酮的得率分别为0.192%和0.088%。 (2)粉红女士苹果类黄酮具有较强的体外抗氧化活性,且呈浓度相关性,果皮类黄酮清除自由基的能力高于果肉类黄酮,对O2·-的清除效果最好。0.8mg/mL的果皮类黄酮对DPPH·和02·-的清除率分别为64.38%和84.84%,总还原力为0.175。 (3)粉红女士苹果类黄酮具有抗肿瘤活性,果皮类黄酮抑癌效果强于果肉类黄酮。果皮、果肉类黄酮对人结肠癌LoVo和乳腺癌MCF-7细胞的增殖抑制率和毒性随着浓度的增加而逐渐增大,100mg/mL的果皮类黄酮对MCF-7细胞增殖的抑制率为57.25%。 (4)粉红女士苹果类黄酮可诱导LoVo及MCF-7细胞产生大量活性氧,促进肿瘤细胞凋亡。 (5)以槲皮素为原料通过磺化反应合成的槲皮素衍生物槲皮素-5’,8-磺酸钠(QS)和槲皮素-4',7-硫酸钠(QOS)具有很好的水溶性,纯度97%,分子结构式均为C15H8Na2013S2,属于同分异构体。 (6)槲皮素及其衍生物对人结肠癌LoVo和乳腺癌MCF-7细胞抗癌活性表现出剂量相关性,对乳腺癌MCF-7细胞系的敏感度高于结肠癌LoVo细胞系。槲皮素、槲皮素-5',8-磺酸钠(QS)、槲皮素-4',7-硫酸钠(QOS)和芦丁四种黄酮类化合物的抗癌活性强弱次序为：槲皮素-5’,8-磺酸钠(QS)槲皮素芦丁槲皮素-4',7-硫酸钠(QOS),100μM的槲皮素-5’,8-磺酸钠对MCF-7细胞的增殖抑制率可达92.38%,能诱导90.66%的肿瘤细胞发生凋亡。
[Abstract]:In this paper, the total flavonoid contents of Pink Lady apple pulp and pericarp were determined, and the antioxidant activity of pink lady apple flavonoids in vitro was studied, based on the grant of Professor Yang Xingbin's National Natural Science Foundation.Two new Quercetin derivatives, Quercetin -5 and Sodium Quercetin 8-Sulfonate, were synthesized from quercetin by sulfonation reaction.Two cell lines, human colon cancer LoVo and breast cancer MCF-7, were used to evaluate the toxicity of flavonoids to tumor cells by MTT assay.The morphological damage of tumor cells was observed by Nikon-TS100 inverted microscope. The morphologic changes of tumor nuclei were observed under inverted fluorescence microscope by Hoechst33258 staining.Flow cytometry was used to analyze the cell cycle distribution and the production of reactive oxygen species in tumor cells.The antitumor activity of quercetin and its derivatives, quercetin -5 and sodium Quercetin 8-sulfonate (QS) and Quercetin -4 (7-Sodium Sulfate) against LoVo and MCF-7 in human colon cancer and breast cancer were studied.The findings are summarized as follows:(1) the contents of flavonoids in peel and pulp of Pink Lady apple were 24.05 卤0.14mg/g and 10.98 卤0.25 mg / g, respectively. The yield of flavonoid was 0.1922% and 0.088%, respectively.(2) the apple flavonoids of Pink Lady had strong antioxidant activity in vitro and showed a concentration-dependent manner. The ability of flavonoids in pericarp to scavenge free radicals was higher than that in pulp.The best scavenging effect for O2-was 0.0.8 mg / mL of pericarp flavonoids, and the removal rates of DPPH and 02- were 64.38% and 84.84%, respectively, and the total reducing power was 0.175.The anticancer activity of apple flavonoids was stronger than that of pulp flavonoids.The inhibitory rate and toxicity of pericarp and pulp flavonoids on the proliferation of human colon cancer LoVo and breast cancer MCF-7 cells increased gradually with the increase of concentration. The inhibitory rate of flavonoids in pericarp on the proliferation of MCF-7 cells was 57.25%.4) flavonoids from pink apple could induce LoVo and MCF-7 cells to produce a large amount of reactive oxygen species and promote apoptosis of tumor cells.(5) Quercetin derivatives, Quercetin derivatives, were synthesized by sulfonation reaction from quercetin (QS) and quercetin (-4) -7- sodium thiosulfate (QSs) and quercetin were synthesized by sulfonation reaction. They have good water solubility, purity 97 and molecular structure of C15H8Na2013S2, which belong to isomers of the same fractions of Quercetin and Quercetin (Quercetin) and Quercetin (Quercetin) and Quercetin (Quercetin).(6) Quercetin and its derivatives showed a dose-dependent response to the anticancer activity of human colon cancer LoVo and breast cancer MCF-7 cells, and the sensitivity of quercetin to breast cancer MCF-7 cell line was higher than that of colon cancer LoVo cell line.Quercetin,The anticancer activity of quercetin, quercetin, quercetin and rutin flavonoids are in the order of quercetin -5tachloro8-sulfonate, quercetin, rutin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, quercetin, and sodium Quercetin.Sodium inhibited the proliferation of MCF-7 cells by 92.38% and induced 90.66% of tumor cells to apoptosis.
【学位授予单位】：陕西师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：S661.1;R151

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