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目的探讨阿托伐他汀对7-酮胆固醇(7-KC)诱导的巨噬细胞内质网应激及细胞凋亡的影响。方法Aop E-/-小鼠左肾动脉和左颈总动脉联合部分结扎建立颈动脉易损斑块模型。采用HE染色方法观察斑块病理学改变,用免疫荧光结合激光扫描共聚焦显微镜技术检测斑块中内质网应激(ER stress)相关蛋白CHOP及磷酸化PERK(p-PERK)的表达。体外培养小鼠巨噬细胞RAW264.7,给予7-KC、H_2O_2或联合阿托伐他汀处理后,蛋白质免疫印迹方法(Western blot)测定ER stress相关蛋白CHOP、p-PERK、XBP-1s及凋亡相关蛋白cleaved caspase-3的表达。结果 Aop E-/-小鼠颈动脉易损斑块局部ER stress相关蛋白CHOP的表达及PERK磷酸化水平明显上调;7-KC可诱导小鼠巨噬细胞ER stress,进而诱导细胞凋亡;同时,氧化应激诱导剂H_2O_2也可通过诱导小鼠巨噬细胞ER stress介导细胞凋亡;而阿托伐他汀可抑制7-KC和H_2O_2诱导的巨噬细胞ER stress及其介导的细胞凋亡。结论ER stress可能参与AS易损斑块的形成;阿托伐他汀可通过减少细胞内氧化应激的水平,减轻巨噬细胞ER stress,从而抑制细胞凋亡。
Objective To investigate the effect of atorvastatin on endoplasmic reticulum stress and apoptosis in macrophages induced by 7-ketochol (7-KC). Methods The left carotid artery and left common carotid artery of Aop E - / - mice were partially ligated to establish a carotid vulnerable plaque model. The pathological changes of plaque were observed by HE staining. The expression of ER stress related protein CHOP and phosphorylated PERK (p-PERK) in plaque were detected by immunofluorescence combined with laser scanning confocal microscopy. Mouse macrophage RAW264.7 cells were cultured in vitro. After treatment with 7-KC, H 2 O 2 or atorvastatin, the ER stress-related proteins CHOP, p-PERK, XBP-1s and apoptosis were detected by Western blot The expression of cleaved caspase-3 protein. Results The expression of ER stress-related protein CHOP and PERK phosphorylation in carotid vulnerable plaque of Apo E - / - mice were significantly up-regulated. 7-KC induced ER stress in mouse macrophages and induced apoptosis. , And oxidative stress inducer H_2O_2 can also induce ER stress-induced apoptosis in mouse macrophages. Atorvastatin can inhibit the ER stress induced by 7-KC and H_2O_2-induced macrophages and the apoptosis induced by atorvastatin Death. Conclusions ER stress may be involved in the formation of vulnerable plaques in AS. Atorvastatin may reduce the level of oxidative stress in cells and reduce the ER stress in macrophages, thereby inhibiting apoptosis.