泰山紫草提取物—乙酰紫草素诱导列腺癌PC3细胞凋亡机制的研究

发布时间：2018-04-11 06:01

本文选题：乙酰紫草素 + PC3细胞　；参考：《泰山医学院》2014年硕士论文

【摘要】：背景与目的泰山紫草为传统中药，是泰山四大名药之一，属硬紫草，其有效成分为多种萘醌类物质。现代药学研究证明紫草具有治疗烧伤、强心、抗炎、抗菌、抗病毒及抗肿瘤等作用。为了进一步研究泰山紫草抗肿瘤机制，本课题组的前期研究从泰山紫草中分离纯化得到了三种主要成分：乙酰紫草素、β-羟基异戊酰紫草素和异丁酰紫草素；发现乙酰紫草素能抑制多种肿瘤细胞系的生长，并可诱导肿瘤细胞凋亡。有关紫草素（软紫草中含量较多的一种萘醌类物质）抗肿瘤的研究近几年已有文献报道，但乙酰紫草素抗肿瘤作用的研究报道较少，其分子机制尚不清楚，，值得研究探讨。本研究以乙酰紫草素为诱导剂，通过MTT法已经筛选出前列腺癌PC3细胞株为乙酰紫草素细胞毒性敏感的细胞株，并从形态学和部分生化指标观察到明显的凋亡特征。进一步研究乙酰紫草素诱导前列腺癌PC3细胞株凋亡的分子机制，为天然新型抗肿瘤药物的开发奠定基础。方法 MTT法检测乙酰紫草素对前列腺癌PC3细胞增殖的影响；流式细胞术检测细胞周期和线粒体膜电位的变化；Annexin-V/PI双染分析凋亡率；实时荧光定量PCR检测凋亡相关基因表达水平；Western blotting检测相关蛋白的表达和激活。结果乙酰紫草素显著抑制PC3细胞的生长，其半数抑制浓度（IC50）为8.59±0.42μM，并使细胞周期阻滞在S期。乙酰紫草素抑制Bcl-2蛋白的表达，促进Bax的表达，引起线粒体膜电位下降，导致细胞色素c释放，进而通过激活内源性凋亡途径诱导PC3细胞死亡。结论乙酰紫草素能抑制PC3细胞增殖，且改变了PC3细胞的周期分布，诱导其凋亡。
[Abstract]:Background and purposeAs a traditional Chinese medicine, Rhizoma Taishan is one of the four famous drugs in Taishan, which belongs to Herba chinensis, and its active ingredient is a variety of naphthoquinones.Modern pharmacological studies have shown that the herb has the effects of treating burn, heart-strengthening, anti-inflammation, anti-bacterial, anti-virus and anti-tumor.In order to further study the anti-tumor mechanism of Rhizoma Taishan, three main components were isolated and purified from Taishan Shikonin: Acetyl Shikonin, 尾 -Hydroxyisopentanyl Shikonin and Isobutylol Shikonin;It was found that Acetyl shikonin could inhibit the growth of many tumor cell lines and induce apoptosis of tumor cells.The studies on the antitumor effect of shikonin (a naphthoquinone with more content) have been reported in recent years, but the anti-tumor effect of Acetylporphyrin is less reported, and its molecular mechanism is not clear, so it is worth studying and discussing.In this study, PC3 cell line of prostate cancer was selected by MTT as inducer, and the cell line was sensitive to cytotoxicity of Acetyl porphyrin, and obvious apoptotic characteristics were observed from morphology and some biochemical indexes.To further study the molecular mechanism of apoptosis induced by Acetotaxin in prostate cancer PC3 cell line, and to lay a foundation for the development of new natural antitumor drugs.MethodMTT assay was used to detect the proliferation of prostate cancer PC3 cells, flow cytometry was used to detect the changes of cell cycle and mitochondrial membrane potential, and Annexin-V / Pi double staining was used to analyze the apoptosis rate.The expression level of apoptosis-related genes was detected by real-time fluorescence quantitative PCR. Western blotting was used to detect the expression and activation of related proteins.ResultAcetyl shikonin significantly inhibited the growth of PC3 cells (IC50) was 8.59 卤0.42 渭 m, and the cell cycle was blocked in S phase.Acetyl shikonin inhibits the expression of Bcl-2 protein, promotes the expression of Bax, decreases mitochondrial membrane potential, and results in the release of cytochrome c, which leads to the death of PC3 cells through activation of endogenous apoptosis pathway.ConclusionAcetyl shikonin can inhibit the proliferation of PC3 cells, change the cell cycle distribution and induce apoptosis of PC3 cells.
【学位授予单位】：泰山医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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