本文選題：CILP2 + 氧化低密度脂蛋白��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：第一部分CILP2表達(dá)上調(diào)對氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成的影響目的:在體外,建立泡沫細(xì)胞模型,探討CILP2表達(dá)上調(diào),是否對氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成產(chǎn)生影響。方法:用佛波酯(PMA)處理THP-1細(xì)胞24小時(shí),誘導(dǎo)成巨噬細(xì)胞。誘導(dǎo)成功后與氧化低密度脂蛋白相互作用24小時(shí),在體外建立泡沫細(xì)胞模型。用腺病毒載體Ad-CILP2感染巨噬細(xì)胞,實(shí)時(shí)熒光定量PCR(RT-PCR)檢測CILP2表達(dá)上調(diào)的效率,油紅染色處理細(xì)胞爬片,觀察CILP2表達(dá)上調(diào),對氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成的影響。結(jié)果:油紅染色結(jié)果顯示泡沫細(xì)胞體外模型構(gòu)建成功。利用Ad-CILP2感染細(xì)胞后,巨噬細(xì)胞內(nèi)CILP2 m RNA水平明顯增高(P0.01),且Ad-CILP2+ox LDL組比Ad-GFP+ox LDL組泡沫細(xì)胞形成明顯增多。結(jié)論:CILP2增加氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成。第二部分CILP2上調(diào)增加氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成過程的機(jī)制研究目的:探討CILP2上調(diào)增加氧化低密度脂蛋白誘導(dǎo)的泡沫細(xì)胞形成過程的可能機(jī)制。方法:用熒光標(biāo)記的氧化型低密度脂蛋白(Dil-ox LDL)作用于巨噬細(xì)胞,熒光倒置顯微鏡觀察對脂質(zhì)的攝取情況。利用RT-PCR檢測介導(dǎo)脂質(zhì)攝取的清道夫受體CD36、LOX-1、SR-A m RNA的表達(dá)。利用western blot檢測CD36蛋白水平的表達(dá)。利用RT-PCR檢測參與膽固醇外流調(diào)節(jié)的ABCA1、ABCG1 m RNA的表達(dá)。用PPAR-γ預(yù)處理細(xì)胞24小時(shí)后,利用RT-PCR檢測CD36、LOX-1 m RNA的表達(dá)。結(jié)果:CILP2增加熒光標(biāo)記的氧化型低密度脂蛋白的攝取。與Ad-GFP+ox LDL組比,Ad-CILP2+ox LDL組CD36 m RNA的表達(dá)明顯升高(P0.01),LOX-1 m RNA的表達(dá)明顯升高(P0.05),但CILP2表達(dá)上調(diào)對SR-A、ABCA1、ABCG1 m RNA的表達(dá)無影響。CILP2使CD36蛋白的表達(dá)增加(P0.01)。巨噬細(xì)胞給予PPAR-γ激動(dòng)劑后,CD36 m RNA的表達(dá)升高(P0.05)。而給予PPAR-γ抑制劑后,CD36 m RNA的表達(dá)降低(P0.05)。用PPAR-γ激動(dòng)劑、抑制劑處理對LOX-1 m RNA的表達(dá)無影響。結(jié)論:CILP2通過PPAR-γ信號通路的激活作用,使巨噬細(xì)胞表面CD36的表達(dá)增加,從而增加脂質(zhì)攝取導(dǎo)致泡沫細(xì)胞的形成增多。
[Abstract]:Part one the effect of up-regulation of CILP2 on the formation of foam cells induced by oxidized low density lipoprotein objective: to establish a foam cell model in vitro and to explore the up-regulation of CILP2 expression. Whether it has an effect on the formation of foam cells induced by oxidized low density lipoprotein (LDL). Methods: THP-1 cells were treated with PMA for 24 hours, and macrophages were induced. The foam cell model was established in vitro after 24 hours of interaction with oxidized low density lipoprotein (LDL). Macrophages were infected with adenovirus vector Ad-CILP2. The efficiency of up-regulation of CILP2 expression was detected by real-time quantitative PCRP-PCRR, and the effect of up-regulation of CILP2 expression on the formation of foam cells induced by oxidized low density lipoprotein (OLDL) was observed by oil red staining. Results: oil red staining showed that foam cell model was successfully constructed in vitro. The level of CILP2 mRNA in macrophages was significantly higher than that in Ad-CILP2 infected cells, and the foam cell formation in Ad-CILP2 ox LDL group was significantly higher than that in Ad-GFP ox LDL group. Conclusion: CILP2 increases the formation of foam cells induced by oxidized low density lipoprotein (LDL). The second part: the mechanism of CILP2 upregulation increasing oxidized low density lipoprotein induced foam cell formation objective: to explore the possible mechanism of CILP2 upregulation increasing oxidized low density lipoprotein induced foam cell formation. Methods: macrophages were treated with fluorescent labeled oxidized low density lipoprotein (LDL) and the uptake of lipid was observed by fluorescence inverted microscope. The expression of SR-A mRNA in scavenger receptor CD36 LOX-1 was detected by RT-PCR. The expression of CD36 protein was detected by western blot. The expression of ABCG 1 mRNA involved in the regulation of cholesterol efflux was detected by reverse transcriptase polymerase chain reaction (RT-PCR). After pretreatment with PPAR- 緯 for 24 hours, the expression of CD36, LOX-1 mRNA was detected by RT-PCR. Results: CILP2 increased uptake of oxidized LDL labeled with fluorescence. Compared with Ad-GFP ox LDL group, the expression of CD36 mRNA in Ad-GFP ox LDL group was significantly higher than that in Ad-GFP ox LDL group. The expression of P0.01LOX-1 m mRNA in Ad-GFP ox LDL group was significantly higher than that in Ad-GFP ox LDL group. However, the up-regulation of CILP2 expression had no effect on the expression of ABCG1 mRNA in SR-A- ABCA1- ABCG1 mRNA. CILP2 increased the expression of CD36 protein. The expression of CD36 mRNA in macrophages increased after PPAR- 緯 agonist administration. However, the expression of CD36 mRNA decreased after PPAR- 緯 inhibitor. Treatment with PPAR- 緯 agonist and inhibitor had no effect on the expression of LOX-1 mRNA. Conclusion the expression of CD36 on macrophages was increased by the activation of PPAR- 緯 signal pathway, and lipid uptake increased, resulting in the formation of foam cells.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R541.4

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