本文選題：內(nèi)毒素　切入點(diǎn)：THP-1源性巨噬細(xì)胞　出處：《南華大學(xué)》2012年碩士論文

【摘要】：目的 以THP-1源性巨噬細(xì)胞為研究對象，經(jīng)內(nèi)毒素處理后，觀察細(xì)胞水平PCSK9、LDLR的變化及其規(guī)律，，以便了解PCSK9是否介導(dǎo)內(nèi)毒素在動脈粥樣硬化發(fā)展中起作用。 方法 用不同濃度的內(nèi)毒素(0,0.1,1,10,100μg/ml)處理經(jīng)160nmol/L佛波酯誘導(dǎo)貼壁的THP-1源性巨噬細(xì)胞不同時間(0,3,6,12,24h)，RT-PCR、Western blot分別檢測PCSK9、LDLR mRNA和蛋白質(zhì)的表達(dá)。應(yīng)用Lipofectamine2000轉(zhuǎn)染80nmol/L PCSK9siRNA進(jìn)入THP-1源性巨噬細(xì)胞后，Western blot檢測LDLR蛋白質(zhì)的表達(dá)以及油紅O染色觀察細(xì)胞內(nèi)脂質(zhì)含量。 結(jié)果 隨著內(nèi)毒素濃度的增加，THP-1源性巨噬細(xì)胞PCSK9mRNA、蛋白質(zhì)的表達(dá)逐漸增多。但是隨著內(nèi)毒素濃度的增加，THP-1源性巨噬細(xì)胞LDLR蛋白質(zhì)表達(dá)逐漸減少；RT-PCR檢測發(fā)現(xiàn)，LDLR mRNA表達(dá)在早期時間(3h和6h)減少，但隨后(12h和24h) LDLR mRNA表達(dá)逐漸增多，到24h表達(dá)量略多于對照組。PCSK9siRNA轉(zhuǎn)染THP-1源性巨噬細(xì)胞后，與LPS單獨(dú)處理組相比，PCSK9siRNA組、PCSK9siRNA+LPS組LDLR蛋白質(zhì)表達(dá)都增多，PCSK9siRNA組表達(dá)最為明顯，其細(xì)胞內(nèi)脂質(zhì)的含量最多。 結(jié)論 1.內(nèi)毒素上調(diào)THP-1源性巨噬細(xì)胞PCSK9mRNA、蛋白質(zhì)的表達(dá)；下調(diào)THP-1源性巨噬細(xì)胞LDLR蛋白質(zhì)的表達(dá)，而對于LDLR mRNA的表達(dá)是可變化的； 2.內(nèi)毒素通過PCSK9下調(diào)THP-1源性巨噬細(xì)胞LDLR的表達(dá)，影響脂質(zhì)的蓄積。
[Abstract]:Purpose. The changes and regularity of THP-1 derived macrophages were observed after endotoxin treatment in order to understand whether endotoxin may play a role in the development of atherosclerosis. Method. THP-1 derived macrophages were treated with different concentrations of endotoxin 0. 1 ~ (1) ~ 1 ~ (10) 渭 g 路ml ~ (-1)) to observe the expression of mRNA and protein in THP-1 derived macrophages induced by 160nmol/L phorbol ester at different times. The expression of mRNA and protein in THP-1 derived macrophages was detected by Lipofectamine2000 transfection of 80nmol/L PCSK9siRNA into THP-1 derived macrophages. The expression of LDLR protein and the lipid content in the cells were observed by oil red O staining. Results. With the increase of endotoxin concentration, the protein expression of THP-1 derived macrophage PCSK9mRNAs gradually increased, but with the increase of endotoxin concentration, the LDLR protein expression of THP-1 derived macrophages decreased gradually. The period time of 3 h and 6 h) decreased, However, the expression of LDLR mRNA increased gradually at 12h and 24h. After transfection of PCSK9siRNA into THP-1 derived macrophages at 24 h, the expression of LDLR protein in PCSK9siRNA group was higher than that in PCSK9siRNA LPS group, and the expression of LDLR protein in PCSK9siRNA group was higher than that in PCSK9siRNA group. The content of lipid in the cells was the highest. Conclusion. 1. Endotoxin up-regulated the expression of PCSK9mRNAs in THP-1 derived macrophages, down-regulated the expression of LDLR protein in THP-1 derived macrophages, and changed the expression of LDLR mRNA. 2. Endotoxin down-regulated the expression of LDLR in THP-1 derived macrophages by PCSK9, and affected the accumulation of lipids.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

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