本文選題：乙酰紫草素 + PC3細(xì)胞��； 參考：《泰山醫(yī)學(xué)院》2014年碩士論文

【摘要】：背景與目的 泰山紫草為傳統(tǒng)中藥，是泰山四大名藥之一，屬硬紫草，其有效成分為多種萘醌類物質(zhì)�，F(xiàn)代藥學(xué)研究證明紫草具有治療燒傷、強(qiáng)心、抗炎、抗菌、抗病毒及抗腫瘤等作用。為了進(jìn)一步研究泰山紫草抗腫瘤機(jī)制，本課題組的前期研究從泰山紫草中分離純化得到了三種主要成分：乙酰紫草素、β-羥基異戊酰紫草素和異丁酰紫草素；發(fā)現(xiàn)乙酰紫草素能抑制多種腫瘤細(xì)胞系的生長(zhǎng)，并可誘導(dǎo)腫瘤細(xì)胞凋亡。有關(guān)紫草素（軟紫草中含量較多的一種萘醌類物質(zhì)）抗腫瘤的研究近幾年已有文獻(xiàn)報(bào)道，但乙酰紫草素抗腫瘤作用的研究報(bào)道較少，其分子機(jī)制尚不清楚，，值得研究探討。本研究以乙酰紫草素為誘導(dǎo)劑，通過MTT法已經(jīng)篩選出前列腺癌PC3細(xì)胞株為乙酰紫草素細(xì)胞毒性敏感的細(xì)胞株，并從形態(tài)學(xué)和部分生化指標(biāo)觀察到明顯的凋亡特征。進(jìn)一步研究乙酰紫草素誘導(dǎo)前列腺癌PC3細(xì)胞株凋亡的分子機(jī)制，為天然新型抗腫瘤藥物的開發(fā)奠定基礎(chǔ)。 方法 MTT法檢測(cè)乙酰紫草素對(duì)前列腺癌PC3細(xì)胞增殖的影響；流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期和線粒體膜電位的變化；Annexin-V/PI雙染分析凋亡率；實(shí)時(shí)熒光定量PCR檢測(cè)凋亡相關(guān)基因表達(dá)水平；Western blotting檢測(cè)相關(guān)蛋白的表達(dá)和激活。 結(jié)果 乙酰紫草素顯著抑制PC3細(xì)胞的生長(zhǎng)，其半數(shù)抑制濃度（IC50）為8.59±0.42μM，并使細(xì)胞周期阻滯在S期。乙酰紫草素抑制Bcl-2蛋白的表達(dá)，促進(jìn)Bax的表達(dá)，引起線粒體膜電位下降，導(dǎo)致細(xì)胞色素c釋放，進(jìn)而通過激活內(nèi)源性凋亡途徑誘導(dǎo)PC3細(xì)胞死亡。 結(jié)論 乙酰紫草素能抑制PC3細(xì)胞增殖，且改變了PC3細(xì)胞的周期分布，誘導(dǎo)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 鹵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.
【學(xué)位授予單位】：泰山醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.25

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