CILP2增加氧化低密度脂蛋白诱导的泡沫细胞形成及机制研究

发布时间：2018-06-18 14:55

本文选题：CILP2 + 氧化低密度脂蛋白　；参考：《重庆医科大学》2017年硕士论文

【摘要】：第一部分CILP2表达上调对氧化低密度脂蛋白诱导的泡沫细胞形成的影响目的:在体外,建立泡沫细胞模型,探讨CILP2表达上调,是否对氧化低密度脂蛋白诱导的泡沫细胞形成产生影响。方法:用佛波酯(PMA)处理THP-1细胞24小时,诱导成巨噬细胞。诱导成功后与氧化低密度脂蛋白相互作用24小时,在体外建立泡沫细胞模型。用腺病毒载体Ad-CILP2感染巨噬细胞,实时荧光定量PCR(RT-PCR)检测CILP2表达上调的效率,油红染色处理细胞爬片,观察CILP2表达上调,对氧化低密度脂蛋白诱导的泡沫细胞形成的影响。结果:油红染色结果显示泡沫细胞体外模型构建成功。利用Ad-CILP2感染细胞后,巨噬细胞内CILP2 m RNA水平明显增高(P0.01),且Ad-CILP2+ox LDL组比Ad-GFP+ox LDL组泡沫细胞形成明显增多。结论:CILP2增加氧化低密度脂蛋白诱导的泡沫细胞形成。第二部分CILP2上调增加氧化低密度脂蛋白诱导的泡沫细胞形成过程的机制研究目的:探讨CILP2上调增加氧化低密度脂蛋白诱导的泡沫细胞形成过程的可能机制。方法:用荧光标记的氧化型低密度脂蛋白(Dil-ox LDL)作用于巨噬细胞,荧光倒置显微镜观察对脂质的摄取情况。利用RT-PCR检测介导脂质摄取的清道夫受体CD36、LOX-1、SR-A m RNA的表达。利用western blot检测CD36蛋白水平的表达。利用RT-PCR检测参与胆固醇外流调节的ABCA1、ABCG1 m RNA的表达。用PPAR-γ预处理细胞24小时后,利用RT-PCR检测CD36、LOX-1 m RNA的表达。结果:CILP2增加荧光标记的氧化型低密度脂蛋白的摄取。与Ad-GFP+ox LDL组比,Ad-CILP2+ox LDL组CD36 m RNA的表达明显升高(P0.01),LOX-1 m RNA的表达明显升高(P0.05),但CILP2表达上调对SR-A、ABCA1、ABCG1 m RNA的表达无影响。CILP2使CD36蛋白的表达增加(P0.01)。巨噬细胞给予PPAR-γ激动剂后,CD36 m RNA的表达升高(P0.05)。而给予PPAR-γ抑制剂后,CD36 m RNA的表达降低(P0.05)。用PPAR-γ激动剂、抑制剂处理对LOX-1 m RNA的表达无影响。结论:CILP2通过PPAR-γ信号通路的激活作用,使巨噬细胞表面CD36的表达增加,从而增加脂质摄取导致泡沫细胞的形成增多。
[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- 纬 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- 纬 agonist administration. However, the expression of CD36 mRNA decreased after PPAR- 纬 inhibitor. Treatment with PPAR- 纬 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- 纬 signal pathway, and lipid uptake increased, resulting in the formation of foam cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.4

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