本文選題：褪黑素 + 胸腺細(xì)胞�。� 參考：《福建醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 目的通過(guò)研究體外培養(yǎng)大鼠胸腺細(xì)胞的自然凋亡和褪黑素對(duì)其凋亡的影響及機(jī)制,探索褪黑素影響胸腺細(xì)胞凋亡的機(jī)制,進(jìn)一步揭示褪黑素與胸腺發(fā)育及細(xì)胞免疫之間的關(guān)系,從而豐富對(duì)神經(jīng)-內(nèi)分泌-免疫網(wǎng)絡(luò)的認(rèn)識(shí)。 方法①體外培養(yǎng)大鼠胸腺細(xì)胞,分為空白對(duì)照組(Ctrl)和褪黑素干預(yù)組M1、M2、M3、M4、M5(藥物濃度分別為10~(-3)、10~(-4)、10~(-6)、10~(-8)、10~(-10)mol/L)。②在培養(yǎng)的第4、8小時(shí)以Annexin-V/PI雙標(biāo)法染色及流式細(xì)胞儀檢測(cè),觀察不同濃度褪黑素干預(yù)下胸腺細(xì)胞凋亡率的變化。③用RT-PCR法及凝膠成像分析系統(tǒng)觀察分析培養(yǎng)的第4、8小時(shí),不同濃度褪黑素干預(yù)下胸腺細(xì)胞Bax、Bcl-xL、Bim mRNA量的變化。④運(yùn)用western blot法觀察培養(yǎng)的第4、8小時(shí),不同濃度褪黑素干預(yù)下胸腺細(xì)胞Bax、Bcl-xL和Bim蛋白表達(dá)量的變化。 結(jié)果①流式細(xì)胞檢測(cè)結(jié)果顯示,培養(yǎng)4小時(shí),FITC~+/ PI~-的大鼠胸腺細(xì)胞高于8小時(shí),相同培養(yǎng)時(shí)間的M1、M2、M3組FITC~+/ PI~-細(xì)胞明顯下降。②半定量PT-PCR及Western blot結(jié)果顯示,Bax和Bim的表達(dá)隨時(shí)間延長(zhǎng)而增高,mRNA與蛋白水平一致,但對(duì)照組與褪黑素干預(yù)組之間無(wú)明顯差異;相同培養(yǎng)時(shí)間的M1、M2、M3組與對(duì)照組相比,Bcl-xL mRNA與蛋白水平均出現(xiàn)升高,但同一處理組的4小時(shí)與8小時(shí)比較無(wú)明顯變化。 結(jié)論體外培養(yǎng)的大鼠胸腺細(xì)胞隨著時(shí)間延長(zhǎng)而凋亡增加,褪黑素能降低胸腺細(xì)胞的早期凋亡率,并呈劑量依賴性。Bax和Bim的表達(dá)隨培養(yǎng)時(shí)間延長(zhǎng)而增高,褪黑素干預(yù)對(duì)其無(wú)明顯影響;褪黑素干預(yù)后Bcl-x_L的表達(dá)水平呈劑量依賴性增高,但不隨培養(yǎng)時(shí)間延長(zhǎng)而變化。因此,褪黑素可通過(guò)影響線粒體途徑從而調(diào)控大鼠胸腺細(xì)胞的凋亡。
[Abstract]:Objective to study the natural apoptosis of rat thymocytes in vitro and the effect and mechanism of melatonin on thymocyte apoptosis, and to explore the mechanism of melatonin affecting thymocyte apoptosis, and to further reveal the relationship between melatonin and thymus development and cellular immunity. So as to enrich the understanding of neural-endocrine-immune network. Methods 1 Rat thymocytes were cultured in vitro and divided into two groups: control group (Ctrl) and melatonin intervention group (M1OM2M3OM4M5). The concentrations of M1OM2M3OM4M5 (the drug concentrations were 10OTr / L5) were determined by Annexin-V/PI double labeling method and flow cytometry at 48 hours after culture. The apoptosis rate of thymocytes was observed at different concentrations of melatonin. 3 the culture was observed by RT-PCR method and gel imaging analysis system at the 4th hour of culture. Changes of Bax-xLnBim mRNA in thymocytes treated with melatonin at different concentrations. The expression of Bax-xL and Bim protein in thymocytes was observed by western blot method at the 8th hour of culture and by different concentrations of melatonin. Results 1 the results of flow cytometry showed that the thymocytes cultured for 4 hours in FITC- / PII- were higher than 8 hours. The expression of Bax and Bim in the FITC- / PII-M3 group decreased significantly with the increase of time, but there was no significant difference between the control group and the melatonin intervention group. The levels of Bcl-xL mRNA and protein in M1M2M3 group were higher than those in control group, but there was no significant change in the same treatment group at 4 hours and 8 hours. Conclusion the apoptosis of rat thymocytes in vitro increases with time. Melatonin can decrease the early apoptosis rate of thymocytes and increase the expression of .Bax and Bim in a dose-dependent manner. The level of Bcl-x_L expression was increased in a dose-dependent manner, but did not change with the prolongation of culture time. Therefore, melatonin can regulate the apoptosis of rat thymocytes by affecting the mitochondrial pathway.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

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