本文選題：乙酰紫草素 + PC3細胞��； 參考：《泰山醫(yī)學院》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�，F(xiàn)代藥學研究證明紫草具有治療燒傷、強心、抗炎、抗菌、抗病毒及抗腫瘤等作用。為了進一步研究泰山紫草抗腫瘤機制，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胞系的生長，并可誘導腫瘤細胞凋亡。有關紫草素（軟紫草中含量較多的一種萘醌類物質）抗腫瘤的研究近幾年已有文獻報道，但乙酰紫草素抗腫瘤作用的研究報道較少，其分子機制尚不清楚，，值得研究探討。本研究以乙酰紫草素為誘導劑，通過MTT法已經(jīng)篩選出前列腺癌PC3細胞株為乙酰紫草素細胞毒性敏感的細胞株，并從形態(tài)學和部分生化指標觀察到明顯的凋亡特征。進一步研究乙酰紫草素誘導前列腺癌PC3細胞株凋亡的分子機制，為天然新型抗腫瘤藥物的開發(fā)奠定基礎。 方法 MTT法檢測乙酰紫草素對前列腺癌PC3細胞增殖的影響；流式細胞術檢測細胞周期和線粒體膜電位的變化；Annexin-V/PI雙染分析凋亡率；實時熒光定量PCR檢測凋亡相關基因表達水平；Western blotting檢測相關蛋白的表達和激活。 結果 乙酰紫草素顯著抑制PC3細胞的生長，其半數(shù)抑制濃度（IC50）為8.59±0.42μM，并使細胞周期阻滯在S期。乙酰紫草素抑制Bcl-2蛋白的表達，促進Bax的表達，引起線粒體膜電位下降，導致細胞色素c釋放，進而通過激活內源性凋亡途徑誘導PC3細胞死亡。 結論 乙酰紫草素能抑制PC3細胞增殖，且改變了PC3細胞的周期分布，誘導其凋亡。
[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.
【學位授予單位】：泰山醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.25

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