本文選題：吳茱萸堿 + 膠質瘤　； 參考：《蘇州大學》2014年碩士論文

【摘要】：膠質瘤是中樞神經系統(tǒng)中最常見的顱內腫瘤，并且有著很高的致死率。與此同時，它有較高的侵襲性且不易于治療。對膠質瘤的各種臨床治療僅僅只能延長患者幾個月的生存期，預后極其不佳。因此，發(fā)掘一個新的抗癌藥物對于膠質瘤的治療至關重要。傳統(tǒng)中藥用于治療腫瘤已有很長一段時間，，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和轉移等多種生物學特性。然而吳茱萸堿對膠質瘤的作用及其作用機制均不明確。 目的研究吳茱萸堿（Evodiamine)對人膠質瘤U251細胞增殖和凋亡的影響及其可能的作用機制。 方法體外培養(yǎng)人膠質瘤U251細胞，并將其分為空白對照組及25、50、100μg.mL-1吳茱萸堿4組。應用MTT法檢測吳茱萸堿在24、48和72h對U251細胞的增殖抑制作用；Hoechst33258熒光染色法檢測吳茱萸堿誘導膠質瘤U251細胞凋亡；采用AnnexinV-FITC/PI雙染法檢測各組24h細胞早期凋亡率；Western blot法分析凋亡相關蛋白的變化。 結果與空白對照組同期比較，25、50、100μg.mL-1吳茱萸堿組生長抑制率在24、48、72h均增加，隨著吳茱萸堿濃度增加和作用時間延長，細胞增殖抑制越明顯，其抑制作用呈現劑量依賴性和時間依賴性，差異有統(tǒng)計學意義(P＜0.05)。Hoechst33258熒光染色顯示吳茱萸堿作用24h后U251細胞出現典型的細胞凋亡特征，各處理組均可見凋亡小體。U251細胞凋亡小體數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間延長，細胞的凋亡率逐漸升高。與空白對照組自發(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)計學意義(P＜0.05)。Westernblot實驗顯示，與空白對照組同期比較，25、50、100μg.mL-1吳茱萸堿上調了Fas、FADD、Caspase-8及Caspase-3蛋白表達，Bcl-2蛋白表達明顯下降，Bax蛋白表達明顯上升，差異均有統(tǒng)計學意義(P＜0.05)。上述指標均呈時間和劑量依賴性。 結論吳茱萸堿對U251細胞具有明顯的抑制細胞增殖和促進細胞凋亡的作用，其機制可能與上調Fas途徑和下調Bcl-2/Bax有關。
[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.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R739.41

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