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目的在高脂饮食诱导小鼠动脉粥样硬化中研究CYP2C8基因对动脉粥样硬化的作用及相关机制。方法 :以20和40周龄APOEKO+/-CYP2C8Tg+/-和CYP2C8Tg+/-转基因的小鼠为研究对象,相同基因背景和周龄的同窝APOEKO+/-和C57BL/6小鼠设为对照。在第5,20,40周时测各组动物EET含量,用Real-time PCR法检测各组小鼠的主动脉PPAR-γ和MCP-1基因表达水平,Western blot分析各组小鼠的主动脉(p-)eNOS和NF-κB蛋白的表达与小鼠的肝脏PI3K、(p-)AKT、(p-)ERK和(p-)P38等信号通路蛋白的表达情况。结果 :在高脂饮食中,CYP2C8Tg+/-组和APOEKO+/-CYP2C8Tg+/-组的EET浓度明显增高。Western blot显示:高表达CYP2C8在肝脏中明显上调PI3K、(p-)AKT、(p-)ERK和(p-)P38蛋白;在主动脉明显上调(p-)eNOS蛋白,下调NF-κB蛋白。Real-time PCR分析显示高表达CYP2C8在主动脉中下调MCP-1蛋白。结论 :本研究高表达CYP2C8基因具有显著的抗炎作用,这一作用是与激活PI3K、(p-)AKT、(p-)ERK、(p-)P38和(p-)eNOS蛋白有关,还与在主动脉中抑制了MCP-1基因有关。
Objective To study the effect and mechanism of CYP2C8 gene on atherosclerosis in atherosclerosis induced by high fat diet in mice. METHODS: Mice of APOEKO +/- CYP2C8Tg +/- and CYP2C8Tg +/- transgenics at 20 and 40 weeks of age were used as controls and littermates APOEKO +/- and C57BL / 6 mice of the same genetic background and age were set as controls. At the 5th, 20th and 40th weeks, the contents of EET in each group were measured. The expression of PPAR-γ and MCP-1 gene in the aorta of each group were detected by Real-time PCR. The expression of arterial (p-) eNOS and NF-κB proteins was correlated with the expression of PI3K, (p-) AKT, (p-) ERK and (p-) P38 in mice liver. Results: The EET concentrations of CYP2C8Tg +/- and APOEKO +/- CYP2C8Tg +/- groups were significantly higher in the high-fat diet. Western blot showed that CYP2C8 overexpression significantly upregulated PI3K, (p-) AKT, (p-) ERK and (p-) P38 in the liver, significantly upregulated (p-) eNOS in the aorta and downregulated NF- KB . Real-time PCR analysis showed that overexpression of CYP2C8 downregulates MCP-1 protein in the aorta. CONCLUSIONS: CYP2C8 gene overexpression has a significant anti-inflammatory effect in this study and is related to the activation of PI3K, (p-) AKT, (p-) ERK, (p-) P38 and (p-) eNOS proteins Is associated with inhibition of the MCP-1 gene in the aorta.