本文關(guān)鍵詞： 內(nèi)源性大麻素系統(tǒng) 膠質(zhì)瘤細胞 雷公藤紅素　出處：《廈門大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景：腦膠質(zhì)瘤是惡性度很高、易復(fù)發(fā)、預(yù)后差的顱內(nèi)惡性腫瘤之一,在臨床上一直是個難題。許多研究證明內(nèi)源性大麻素具有抗腫瘤生成作用,包括抑制增殖、誘導(dǎo)凋亡和抑制遷移作用,但其對膠質(zhì)瘤的作用靶點及詳細機制尚未完全闡明。雷公藤紅素可以抑制蛋白酶體活性、阻滯在細胞周期和誘導(dǎo)細胞凋亡,這使得它成為一種潛在的治療膠質(zhì)瘤的藥物。 目的：本研究通過觀察利用單獨分子生物學(xué)、分子藥物學(xué)手段調(diào)控內(nèi)源性大麻素系統(tǒng)或與雷公藤紅素聯(lián)用后不同膠質(zhì)瘤細胞的增殖情況,在細胞水平探討膠質(zhì)瘤可能的治療靶點,為雷公藤紅素開發(fā)成為抗膠質(zhì)瘤新藥提供依據(jù)。 方法：用細胞計數(shù)板計算細胞數(shù)繪制細胞的生長曲線,根據(jù)曲線計算出倍增時間；用Western blot方法檢測細胞內(nèi)CD133表達；內(nèi)源性大麻素、游離脂肪酸及神經(jīng)酰胺采用LC-MS/MS方法檢測;使用熒光定量PCR (RT-qPCR)技術(shù)檢測大麻素受體CB1、CB2,大麻素水解酶FAAH、MGL等的基因mRNA表達水平及雷公藤紅素對其的影響；用CCK-8試劑盒檢測單獨調(diào)控內(nèi)源性大麻素系統(tǒng)或與雷公藤紅素聯(lián)用后膠質(zhì)瘤細胞增殖的影響。 結(jié)果：SHG細胞增長速度最快,具有干細胞特征。四種細胞內(nèi)FFAs、Ceramides含量均有顯著性差異。內(nèi)源性大麻素系統(tǒng)中大麻素AEA、2-AG及其水解酶FAAH、MGL,以及CB1的表達有顯著性差異,2-AG和MGL是調(diào)控內(nèi)源性大麻素系統(tǒng)的重要靶點。雷公藤紅素對膠質(zhì)瘤細胞具有一定程度的抑制作用,存在時間和劑量依賴性,在此過程中上調(diào)了大麻素受體的基因表達,CB2的升高更為明顯,雷公藤紅素與2-AG分解酶抑制劑JZL184、小劑量的大麻素受體激動劑都可以增強的抗膠質(zhì)瘤作用,可以作為抗膠質(zhì)瘤作用的聯(lián)合用藥開發(fā)。 結(jié)論：上調(diào)內(nèi)源性大麻素2-AG含量或激活相關(guān)信號通道會抑制膠質(zhì)瘤細胞的增殖,內(nèi)源性大麻素系統(tǒng)是治療膠質(zhì)瘤的有效靶點,雷公藤紅素作為抗膠質(zhì)瘤聯(lián)合用藥開發(fā)具有良好的臨床應(yīng)用前景。
[Abstract]:Background: glioma is one of intracranial malignant tumors with high degree of malignancy, easy recurrence and poor prognosis, which has been a difficult problem in clinic. Many studies have proved that endogenous cannabinoids have anti-tumorigenic effects, including inhibition of proliferation. However, the target and detailed mechanism of its action on glioma has not been fully elucidated. Tripterygium wilfordii can inhibit proteasome activity, block cell cycle and induce apoptosis. This makes it a potential therapy for glioma. Objective: to investigate the effects of monolayer molecular biology and molecular pharmacology on the proliferation of glioma cells with endogenous cannabinoid system or tripterygium wilfordii. To explore the possible therapeutic targets of glioma at cell level and to provide evidence for the development of tripterine as a new anti-glioma drug. Methods: the cell growth curve was calculated by cell counting board, and the doubling time was calculated according to the curve. The expression of CD133 was detected by Western blot method, and the endogenous cannabinoid, free fatty acid and ceramide were detected by LC-MS/MS method. The level of mRNA expression of cannabinoid receptor CB1 (CB1) and cannabinase (FAAHH) MGL and the effect of tripterine on it were detected by fluorescence quantitative PCR RT-qPCR technique. The effects of endogenous cannabinoid system alone or tripterygium wilfordii on the proliferation of glioma cells were detected by CCK-8 kit. The results showed that the growth rate of the cell was the fastest. There were significant differences in the contents of FFAs-Ceramides in four kinds of cells. The expression of cannabinoid AEA-2-AG and its hydrolase FAAHMAMGLand CB1 in endogenous cannabinoid system were significantly different, and 2-AG and MGL regulated the endogenous cannabinoid system. Tripterygium wilfordii has a certain degree of inhibitory effect on glioma cells. In a time-and dose-dependent manner, the gene expression of cannabinoid receptor (CB2) was upregulated in a dose-dependent manner. Tripterygium wilfordii and 2-AG decomposing enzyme inhibitor JZL1844, a small dose of cannabinoid receptor agonist, could enhance the anti-glioma effect. It can be used as a combined drug for anti-glioma. Conclusion: upregulation of endogenous cannabinoid 2-AG or activation of related signal channels can inhibit the proliferation of glioma cells. Endogenous cannabinoid system is an effective target for the treatment of glioma. Tripterygium wilfordii as a combined antiglioma drug has a good clinical application prospects.
【學(xué)位授予單位】：廈門大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.4

【共引文獻】

相關(guān)期刊論文 前9條

1 浦毅;喻永濤;陳翔;孫利華;羊正祥;;多西紫杉醇聯(lián)合環(huán)巴胺抑制腦膠質(zhì)瘤生長的實驗研究[J];第二軍醫(yī)大學(xué)學(xué)報;2013年12期

2 劉哲言;盧一鳴;屈武斌;張成崗;;人類管家基因和非管家基因microRNA綁定密度的比較及與3'UTR進化保守性關(guān)系的分析[J];軍事醫(yī)學(xué);2013年11期

3 賀娟;凌宏艷;胡弼;;脂肪水解與脂肪信號[J];中南醫(yī)學(xué)科學(xué)雜志;2014年02期

4 梁曉陽;劉洛同;明揚;陳禮剛;;人腦膠質(zhì)瘤中Fas、Caspase-8的表達[J];海南醫(yī)學(xué);2014年07期

5 程瑩;潘長玉;;Rimonabant:一種有希望的代謝綜合征治療藥物[J];中國藥物應(yīng)用與監(jiān)測;2007年03期

6 孟祥龍;廖申權(quán);戚南山;吳彩艷;呂敏娜;李娟;孫銘飛;李國清;;柔嫩艾美耳球蟲MAGL基因的克隆及表達[J];畜牧與獸醫(yī);2013年09期

7 王中奎;李燕婷;趙東升;韓大雄;;新型脂肪酰胺水解酶抑制劑的設(shè)計、合成和活性評價[J];中國現(xiàn)代應(yīng)用藥學(xué);2014年01期

8 張?zhí)N蘊;周偉;孫卯;黃鑼;陳曉品;;Clusterin基因沉默對腦膠質(zhì)瘤U87細胞放射敏感性的影響[J];重慶醫(yī)科大學(xué)學(xué)報;2014年02期

9 張自力;張衍;張涉;段旭酉;郭瑤;倪雯霞;王妤清;鄭仕中;;大麻素受體在肝纖維化及肝硬化中的作用及研究進展[J];中國藥理學(xué)通報;2014年03期

相關(guān)博士學(xué)位論文 前10條

1 原志慶;TFPI-2基因CpG島在甲狀腺結(jié)節(jié)性疾病中的甲基化狀態(tài)及意義[D];鄭州大學(xué);2013年

2 季天海;LATS1對膠質(zhì)瘤細胞增殖和侵襲的影響與膠質(zhì)瘤細胞代謝組學(xué)研究[D];第三軍醫(yī)大學(xué);2013年

3 杜璐;中腦導(dǎo)水管周圍灰質(zhì)HCN通道參與大鼠神經(jīng)病理性痛的機制研究[D];第三軍醫(yī)大學(xué);2013年

4 劉昌燕;大草蛉卵黃原蛋白基因功能研究[D];中國農(nóng)業(yè)科學(xué)院;2013年

5 孫哲;手術(shù)聯(lián)合放射性~(125)I粒子植入治療腦惡性腫瘤的臨床應(yīng)用研究[D];新疆醫(yī)科大學(xué);2013年

6 馬R，

本文編號：1502240