本文選題：吳茱萸堿 + 膠質(zhì)瘤�。� 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：膠質(zhì)瘤是中樞神經(jīng)系統(tǒng)中最常見(jiàn)的顱內(nèi)腫瘤，并且有著很高的致死率。與此同時(shí)，它有較高的侵襲性且不易于治療。對(duì)膠質(zhì)瘤的各種臨床治療僅僅只能延長(zhǎng)患者幾個(gè)月的生存期，預(yù)后極其不佳。因此，發(fā)掘一個(gè)新的抗癌藥物對(duì)于膠質(zhì)瘤的治療至關(guān)重要。傳統(tǒng)中藥用于治療腫瘤已有很長(zhǎng)一段時(shí)間，，大量研究表明許多中藥的提取物或者他們的混合物在體內(nèi)外都有抗癌作用。吳茱萸堿是從吳茱萸的中提取的主要生物堿，它具有抑制腫瘤生長(zhǎng)和轉(zhuǎn)移等多種生物學(xué)特性。然而吳茱萸堿對(duì)膠質(zhì)瘤的作用及其作用機(jī)制均不明確。 目的研究吳茱萸堿（Evodiamine)對(duì)人膠質(zhì)瘤U251細(xì)胞增殖和凋亡的影響及其可能的作用機(jī)制。 方法體外培養(yǎng)人膠質(zhì)瘤U251細(xì)胞，并將其分為空白對(duì)照組及25、50、100μg.mL-1吳茱萸堿4組。應(yīng)用MTT法檢測(cè)吳茱萸堿在24、48和72h對(duì)U251細(xì)胞的增殖抑制作用；Hoechst33258熒光染色法檢測(cè)吳茱萸堿誘導(dǎo)膠質(zhì)瘤U251細(xì)胞凋亡；采用AnnexinV-FITC/PI雙染法檢測(cè)各組24h細(xì)胞早期凋亡率；Western blot法分析凋亡相關(guān)蛋白的變化。 結(jié)果與空白對(duì)照組同期比較，25、50、100μg.mL-1吳茱萸堿組生長(zhǎng)抑制率在24、48、72h均增加，隨著吳茱萸堿濃度增加和作用時(shí)間延長(zhǎng)，細(xì)胞增殖抑制越明顯，其抑制作用呈現(xiàn)劑量依賴性和時(shí)間依賴性，差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)。Hoechst33258熒光染色顯示吳茱萸堿作用24h后U251細(xì)胞出現(xiàn)典型的細(xì)胞凋亡特征，各處理組均可見(jiàn)凋亡小體。U251細(xì)胞凋亡小體數(shù)量隨著吳茱萸堿濃度的增加而逐漸增多，且碎裂的顆粒逐漸變小。隨著吳茱萸堿濃度增加和作用時(shí)間延長(zhǎng)，細(xì)胞的凋亡率逐漸升高。與空白對(duì)照組自發(fā)早期凋亡率3.12%比較，25、50、100μg.mL-1吳茱萸堿組早期凋亡率分別為8.65%、19.47%及28.97%，差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05)。Westernblot實(shí)驗(yàn)顯示，與空白對(duì)照組同期比較，25、50、100μg.mL-1吳茱萸堿上調(diào)了Fas、FADD、Caspase-8及Caspase-3蛋白表達(dá)，Bcl-2蛋白表達(dá)明顯下降，Bax蛋白表達(dá)明顯上升，差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05)。上述指標(biāo)均呈時(shí)間和劑量依賴性。 結(jié)論吳茱萸堿對(duì)U251細(xì)胞具有明顯的抑制細(xì)胞增殖和促進(jìn)細(xì)胞凋亡的作用，其機(jī)制可能與上調(diào)Fas途徑和下調(diào)Bcl-2/Bax有關(guān)。
[Abstract]:Glioma is the most common intracranial tumor in the central nervous system and has a high mortality. At the same time, it is highly invasive and difficult to treat. The clinical treatment of glioma can only prolong the survival of patients for a few months, and the prognosis is extremely poor. Therefore, the discovery of a new anti-cancer drug is essential for the treatment of gliomas. Traditional Chinese medicine has been used to treat cancer for a long time. A lot of studies have shown that many extracts of traditional Chinese medicine or their mixture have anticancer effect in vivo and in vitro. Rutaecarpine is the main alkaloid extracted from Evodia rutaecarpa. It has many biological characteristics such as inhibiting tumor growth and metastasis. However, the effect of rutaecarpine on glioma and its mechanism are not clear. Aim to study the effect of Evodiamine on proliferation and apoptosis of human glioma U251 cells and its possible mechanism. Methods Human glioma U251 cells were cultured in vitro and divided into control group and 2550100 渭 g.mL-1 rutaecarpine group. The inhibitory effect of evodiamine on U251 cells was detected by MTT method. Hoechst33258 fluorescence staining was used to detect the apoptosis of U251 glioma cells induced by evodiamine. The changes of apoptosis-related proteins were detected by AnnexinV-FITC/PI double staining and Western blot assay. Results compared with the control group, the growth inhibition rate of Evodia officinalis 2550100 渭 g.mL-1 group increased at 24: 48 h. With the increase of Evodia rutaecarpine concentration and the prolongation of the action time, the cell proliferation inhibition was more obvious, and the inhibitory effect was in a dose-and time-dependent manner. The difference was statistically significant (P < 0.05).Hoechst33258). The typical apoptotic characteristics of U251 cells were observed after Evodipine treatment for 24 hours. The number of apoptotic corpuscles in U251 cells increased with the increase of Evodia rutaecarine concentration in all treatment groups. And the broken particles gradually become smaller. With the increase of Evodia officinalis concentration and the prolongation of the action time, the apoptosis rate increased gradually. Compared with the control group, the spontaneous early apoptosis rate was 3.12%. The early apoptotic rate was 8.65%, 19.47% and 28.97% respectively in the 25 ~ 50100 渭 g.mL-1 rutaecarpine group. The difference was statistically significant (P < 0.05).Westernblot). Compared with the control group, Evodia rutaecarcinae 2550100 渭 g.mL-1 upregulated the expression of Caspase-8 and Caspase-3 protein of Fas-FADDN, and the expression of Bax protein increased significantly (P < 0.05). All the above indexes were in a time and dose dependent manner. Conclusion rutaecarpine can inhibit cell proliferation and promote cell apoptosis in U251 cells, and its mechanism may be related to up-regulation of Fas pathway and down-regulation of Bcl-2/Bax.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

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