本文關(guān)鍵詞： 內(nèi)源性大麻素系統(tǒng) 膠質(zhì)瘤細(xì)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(gè)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(duì)膠質(zhì)瘤的作用靶點(diǎn)及詳細(xì)機(jī)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(xì)胞周期和誘導(dǎo)細(xì)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(guò)觀察利用單獨(dú)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(xì)胞的增殖情況,在細(xì)胞水平探討膠質(zhì)瘤可能的治療靶點(diǎn),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(xì)胞計(jì)數(shù)板計(jì)算細(xì)胞數(shù)繪制細(xì)胞的生長(zhǎng)曲線,根據(jù)曲線計(jì)算出倍增時(shí)間；用Western blot方法檢測(cè)細(xì)胞內(nèi)CD133表達(dá)；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(cè);使用熒光定量PCR (RT-qPCR)技術(shù)檢測(cè)大麻素受體CB1、CB2,大麻素水解酶FAAH、MGL等的基因mRNA表達(dá)水平及雷公藤紅素對(duì)其的影響；用CCK-8試劑盒檢測(cè)單獨(dú)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(xì)胞增殖的影響。 結(jié)果：SHG細(xì)胞增長(zhǎng)速度最快,具有干細(xì)胞特征。四種細(xì)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(dá)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(diǎn)。雷公藤紅素對(duì)膠質(zhì)瘤細(xì)胞具有一定程度的抑制作用,存在時(shí)間和劑量依賴性,在此過(guò)程中上調(diào)了大麻素受體的基因表達(dá),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(dòng)劑都可以增強(qiáng)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(hào)通道會(huì)抑制膠質(zhì)瘤細(xì)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(diǎn),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(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.4

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6 馬R，

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