本文選題：氧化低密度脂蛋白 + DC2.4　； 參考：《廣州醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的 觀察氧化低密度脂蛋白（oxidized low-density lipoprotein，ox-LDL）對(duì)小鼠樹突狀細(xì)胞株DC2.4細(xì)胞形態(tài)、表型成熟、膜蛋白CD47及microRNA-155表達(dá)的影響，初步探討CD47和microRNA-155在動(dòng)脈粥樣硬化（atherosclerosis，AS）條件下在DC成熟和遷移中的潛在作用。 方法 1. DC2.4接種于6孔板中，用含10%胎牛血清的RPMI1640完全培養(yǎng)液培養(yǎng)至第3天，改用無血清培養(yǎng)基培養(yǎng)并分為2組，實(shí)驗(yàn)組加入ox-LDL（100ug/mL），對(duì)照組加入磷酸緩沖液（PBS）。使用光學(xué)顯微鏡觀察細(xì)胞形態(tài)學(xué)變化；分別于干預(yù)后12小時(shí)、24小時(shí)收集DC2.4細(xì)胞，采用流式細(xì)胞儀檢測(cè)細(xì)胞表型（MHC-II，CD80和CD86）的表達(dá)。 2. DC2.4接種于6孔板中，用含10%胎牛血清的RPMI1640完全培養(yǎng)液培養(yǎng)至第3天，用無血清培養(yǎng)基培養(yǎng)并分為2組，實(shí)驗(yàn)組加入ox-LDL（100ug/mL），對(duì)照組加入磷酸緩沖液（PBS）。分別于干預(yù)后12小時(shí)、24小時(shí)收集DC2.4細(xì)胞，分別提取蛋白后采用Western blot方法測(cè)定細(xì)胞表面CD47分子的表達(dá)；提取RNA后采用熒光定量PCR方法測(cè)定DC2.4細(xì)胞中microRNA-155的表達(dá)。 結(jié)果 1.1DC2.4形態(tài)學(xué)觀察：對(duì)照組細(xì)胞形態(tài)呈圓形或類圓形、較小、透亮，周邊少量毛刺狀或棘狀突起；ox-LDL刺激12h及24h后觀察細(xì)胞，發(fā)現(xiàn)細(xì)胞體積增大，胞漿逐漸豐富，呈不規(guī)則多型狀態(tài)，具有典型的樹突狀突起。 1.2DC2.4成熟表型變化：收集實(shí)驗(yàn)組和對(duì)照組細(xì)胞進(jìn)行流式細(xì)胞檢測(cè)，經(jīng)過ox-LDL處理后的DC2.4表型（MHC-II，CD80和CD86）的表達(dá)明顯增加，與PBS對(duì)照組相比，存在統(tǒng)計(jì)學(xué)差異（P＜0.05）。 2.1經(jīng)過ox-LDL處理后的DC2.4CD47的表達(dá)下降，與PBS對(duì)照組相比，存在統(tǒng)計(jì)學(xué)差異（P＜0.05）。 2.2經(jīng)過ox-LDL處理后的DC2.4microRNA-155的表達(dá)增高，，與PBS對(duì)照組相比，存在統(tǒng)計(jì)學(xué)差異（P＜0.05）。 結(jié)論 1.ox-LDL可使DC2.4表面類分子MHC-II，CD80和CD86高表達(dá)，誘導(dǎo)DC2.4成熟。 2. ox-LDL可誘導(dǎo)DC2.4表面CD47分子下調(diào)表達(dá)、microRNA-155上調(diào)表達(dá)。
[Abstract]:Purpose To observe the effects of oxidized low density lipoprotein (low-density) on the morphology, phenotypic maturation, membrane protein CD47 and microRNA-155 expression of mouse dendritic cell line DC2.4, and to explore the potential role of CD47 and microRNA-155 in DC maturation and migration under the condition of atherosclerotic atherosclerosis. Method 1. DC2.4 was inoculated in 6-well plate and cultured in 10% fetal bovine serum RPMI1640 for 3 days, then cultured in serum-free medium and divided into two groups. The experimental group was added ox-LDLN 100ugmLX, and the control group was added phosphoric acid buffer solution (PBS). DC2.4 cells were collected at 12 hours and 24 hours after intervention, and the expression of MHC-IIP CD80 and CD86 were detected by flow cytometry. 2. DC2.4 was inoculated in a 6-well plate and cultured in 10% fetal bovine serum RPMI1640 for 3 days. The RPMI1640 was cultured in serum-free medium and divided into two groups. The experimental group was added ox-LDLN 100ugmLX, and the control group was added phosphoric acid buffer solution (PBS). DC2.4 cells were collected at 12 hours and 24 hours after intervention. The expression of CD47 on the cell surface was measured by Western blot method after protein extraction, and the expression of microRNA-155 in DC2.4 cells was detected by fluorescence quantitative PCR after RNA extraction. Result Morphological observation of 1.1DC2.4: the cells in the control group were round or round, small and bright, and a small amount of burrs or spikes around the cells were observed after 12 h and 24 hours of stimulation. It was found that the size of the cells increased, the cytoplasm was gradually rich, and the cells were irregular and polytype. Having typical dendritic protuberances. Changes of mature phenotype of 1.2DC2.4: the expression of MHC-IIP CD80 and CD86 in DC2.4 treated with ox-LDL was significantly increased, compared with that in control group (P < 0.05), and the expression of MHC-IIP CD80 and CD86 was significantly increased after ox-LDL treatment (P < 0.05). 2.1 the expression of DC2.4CD47 decreased after ox-LDL treatment, and there was significant difference between the two groups (P < 0.05). 2.2 the expression of DC2.4microRNA-155 was increased after ox-LDL treatment, and there was significant difference between the two groups (P < 0.05). Conclusion 1.ox-LDL could induce the expression of MHC-IIP CD80 and CD86 on the surface of DC2.4 and induce DC2.4 maturation. 2. Ox-LDL could induce down-regulation of CD47 expression on DC2.4 surface and upregulate the expression of microRNA-155.
【學(xué)位授予單位】：廣州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;OXIDIZED HIGH-DENSITY LIPOPROTEIN PROMOTES MATURATION AND MIGRATION OF BONE MARROW DERIVED DENDRITIC CELLS FROM C57BL/6J MICE[J];Chinese Medical Sciences Journal;2008年04期

本文編號(hào)：1927397