摘要
目的探讨人主动脉内皮细胞(HAEC)在缺血缺氧刺激下内质网应激(ERS)标志蛋白C/EBP同源蛋白10(CHOP-10)的表达变化及意义,并观察阿托伐他汀对上述过程的影响。方法将传代培养的HAEC分为正常对照组、缺血缺氧组、缺血缺氧+CHOP-10基因沉默组以及缺血缺氧+阿托伐他汀组(0.l mol/L、1.0 mol/L、10.0mol/L),缺血缺氧+CHOP-10基因沉默组采用CHOP-10 shRNA下调CHOP-10的基因表达;24 h后采用RT-PCR法检测细胞中CHOP-10的基因表达,Western blot法检测CHOP-10、Caspase-3和Caspase-8的蛋白水平;采用ELISA法检测细胞培养液中白细胞介素6(IL-6)及肿瘤坏死因子α(TNF-α)的浓度;采用CCK8法测定细胞增殖活力。结果与正常对照组相比,HAEC在缺血缺氧损伤时CHOP-10表达明显升高(P<0.01),细胞分泌炎性因子IL-6及TNF-α增加(P<0.01),凋亡蛋白Caspase-3和Caspase-8表达增加(P<0.01),细胞增殖活力明显下降(P<0.01)。阿托伐他汀能呈浓度依赖性地抑制HAEC CHOP-10表达,而缺血缺氧+CHOP-10基因沉默组或缺血缺氧+阿托伐他汀组,炎性介质IL-6、TNF-α的分泌也相应减少,细胞凋亡下降,增殖活力明显增加(P<0.01)。结论缺血缺氧损伤可引起血管内皮细胞发生ERS及炎症反应,导致细胞增殖活力下降,凋亡增加,CHOP-10基因沉默或应用阿托伐他汀干预可减轻缺血缺氧时ERS及炎症反应而对血管内皮细胞产生保护作用。
Aim To investigate the expression of endoplasmic reticulum stress（ ESR） marker C/EBP homologous protein-10（ CHOP-10） in the human aortic endothelial cells（ HAEC） under the ischemia and hypoxia stress and to study the effects of atorvastatin on the process. Methods The cultured HAEC were divided into normal control group,ischemia / hypoxia model group,ischemia / hypoxia plus CHOP-10 shRNA group and ischemia / hypoxia plus atorvastatin treatment group（ 0. 1 mol / L,1. 0 mol / L,10. 0 mol / L）. After 24 hours,the expression of CHOP-10 was detected by RT-PCR and the protein levels of CHOP-10,caspase-3 and caspase-8 were measured by Western blot. The levels of interleukin-6（ IL-6） and tumor necrosis factor-α（ TNF-α） in the supernatant of cell culture medium were measured by ELISA. The CCK-8 test was used to measure cell proliferation. Results As compared with that of the control group,the CHOP-10 expression increased significantly in HAEC under the ischemia and hypoxia stress（ P〈0. 01）,the IL-6 andTNF-α level in the supernatant of cell culture medium were significantly higher than those of the control group（ P〈0. 01）.And the cells apoptosis increased and proliferation activity decreased significantly under the ischemia and hypoxia stress（ P〈0. 01）. The expression of CHOP-10 can be inhibited by atorvastatin in a dose-dependent manner in HAEC under the ischemia and hypoxia stress（ P〈0. 01）. Knock down CHOP-10 expression or application of atorvastatin treatment can inhibit cell apoptosis and increase proliferation activity in the cells of the ischemia / hypoxia group（ P〈0. 01）. Conclusions These findings indicate that ischemia and hypoxia can induce ESR and inflammation in HAEC. CHOP-10 expression up-regulated by ESR can increase cell apoptosis and decrease proliferation activity. Down-regulated CHOP-10 expression or application of atorvastatin treatment can reduce ESR under the ischemia and hypoxia stress and have a protective effect on vascular endothelial cells.
出处
《中国动脉硬化杂志》
CAS
北大核心
2016年第3期245-250,共6页
Chinese Journal of Arteriosclerosis
基金
湖北省宜昌市科技攻关项目(A14301-02)
