蛋白激酶C活性变化对adipophilin介导脂质蓄积的影响

发布时间：2018-03-31 10:53

本文选题：THP-1巨噬细胞　切入点：氧化低密度脂蛋白　出处：《南华大学》2006年硕士论文

【摘要】：胆固醇等中性脂质的蓄积是动脉粥样硬化泡沫细胞形成和四期病变发生的核心环节。在真核细胞中，这些蓄积的脂质主要以脂滴的形式贮存于胞浆中。Adipophilin就是脂滴外周40余种相关蛋白中含量最高的一种。其功能主要是促进细胞中脂质的蓄积，抑制胆固醇的外流。在adipophilin基因启动子区有过氧化物酶体增殖物激活受体PPAR各亚型的反应元件PPRE，能够与PPAR_(γ1)、PPAR_(γ2)结合并调控人adipophilin基因的表达。而参与多种脂质代谢靶基因调控的核转录因子PPARs可以和转录共激活物PBP作用，直接接受信号转导关键酶PKC的磷酸化调控。研究adipophilin在脂质蓄积中的调控因素尤其是具有良好药物开发价值的PKC信号转导通路，有可能为动脉粥样硬化性疾病的防治带来新的契机。本研究首先在动脉粥样硬化新西兰白兔模型上观察Adipophilin和PKC_α的表达；然后在细胞水平上观察oxLDL对A10大鼠主动脉平滑肌细胞、THP-1人巨噬细胞和ECV304人脐静脉内皮细胞PKC活性和PPAR_γ与Adipophilin表达的影响；进一步应用PKC激动剂PMA和抑制剂CalphostinC观察PKC活性的改变对荷脂THP-1巨噬细胞中PKC_α、PPAR_γ和Adipophilin表达的影响，并检测细胞内脂质蓄积的改变情况，以初步探讨信号转导途径中的关键酶PKC调控adipophilin及脂质蓄积的可能机制。第一部分动脉粥样硬化新西兰白兔Adipophilin和PKC_α表达的变化目的在动脉粥样硬化新西兰白兔动物模型上，观察Adipophilin和PKC_α表达的变化。方法采用本课题组建立的新西兰兔动脉粥样硬化动物模型。24只新西兰白兔，，随机分为对照组和实验组。用高胆固醇饮食饲喂新西兰白兔12w，动脉切片HE染色；采用Westernblot蛋白印迹和免疫组化染色检测Adipophilin和PKC_α表达。
[Abstract]:The accumulation of neutral lipid, such as cholesterol, is the core of the formation of atherosclerotic foam cells and the development of stage four lesions.In eukaryotic cells, these accumulated lipids are mainly stored in the cytoplasm in the form of lipid droplets. Adipophilin is one of the highest concentrations of more than 40 proteins associated with lipid droplets.Its function is mainly to promote the accumulation of lipid in cells, inhibit the outflow of cholesterol.In the promoter region of adipophilin gene, PPRE, a response element of peroxisome proliferator activated receptor (PPAR) subtype, can bind to and regulate the expression of human adipophilin gene.Nuclear transcription factor (PPARs), which is involved in the regulation of multiple target genes of lipid metabolism, can interact with the transcription co-activator (PBP) and directly accept the phosphorylation of PKC, the key enzyme of signal transduction.The study of the regulatory factors of adipophilin in lipid accumulation, especially the PKC signal transduction pathway with good drug development value, may bring a new opportunity for the prevention and treatment of atherosclerotic diseases.In this study, the expression of Adipophilin and PKCa was observed in atherosclerotic New Zealand white rabbit model.Then the effects of oxLDL on the activity of PKC and the expression of PPAR- 纬 and Adipophilin in THP-1 human macrophages and ECV304 human umbilical vein endothelial cells (HUVECs) of A10 rat aortic smooth muscle cells were observed at the cellular level.PKC agonist PMA and inhibitor CalphostinC were used to observe the effect of PKC activity on the expression of PKC _ 伪 -PPAR纬 and Adipophilin in cultured THP-1 macrophages, and the changes of intracellular lipid accumulation were detected.To explore the possible mechanism of regulating adipophilin and lipid accumulation by PKC, a key enzyme in signal transduction pathway.Changes of Adipophilin and PKC _ 伪 expression in New Zealand White Rabbits with AtherosclerosisObjective to observe the expression of Adipophilin and PKC _ 伪 in atherosclerotic New Zealand white rabbits.Methods Twenty-four New Zealand white rabbits were randomly divided into two groups: control group and experimental group.New Zealand white rabbits were fed with high cholesterol diet for 12 weeks. The expression of Adipophilin and PKCa was detected by Westernblot Western blot and immunohistochemical staining.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

【相似文献】