摘要
目的 观察高脂损伤冠状动脉内皮细胞早期生长反应因子3(Egr3)表达变化,探讨Egr3对高脂诱导冠状动脉内皮细胞炎症及凋亡的影响.方法 对数生长期人冠状动脉内皮细胞分为对照组(正常培养)、高脂损伤组(氧化低密度脂蛋白处理)、Egr3空载组(siRNA-NC慢病毒转染+氧化低密度脂蛋白处理)、Egr3敲低组(siRNA-Egr3慢病毒转染+氧化低密度脂蛋白处理).转染后取4组细胞,采用实时荧光定量PCR法检测细胞Egr3 mRNA相对表达量,采用Western blot法检测Egr3、caspase-3蛋白相对表达量,采用ELISA法检测细胞上清液肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平,采用CCK-8法检测细胞凋亡率.结果 高脂损伤组、Egr3空载组细胞Egr3 mRNA相对表达量(1.56±0.48、1.53±0.56),Egr3(1.65±0.35、1.55±0.45)、caspase-3(2.43±0.43、2.36±0.29)蛋白相对表达量,细胞上清液 TNF-α[(55.33±4.76)、(57.85±2.92)ng/L]、IL-6[(36.34±1.60)、(31.69±1.83)ng/L]水平及细胞凋亡率[(186.08±20.84)％、(174.34±29.98)％]均高于对照组[1.04±0.20、0.51±0.26、1.60±0.22、(46.10±1.52)ng/L、(23.35±1.69)ng/L、(100.01±8.64)％]、Egr3 敲低组[0.16±0.03、0.23±0.07、1.05±0.29、(33.67±3.26)ng/L、(19.75±1.54)ng/L、(12.24±7.51)％](P＜0.05),对照组均高于 Egr3 敲低组(P＜0.05),高脂损伤组与 Egr3 空载组比较差异均无统计学意义(P＞0.05).结论 高脂可诱导冠状动脉内皮细胞炎症和凋亡,可能与Egr3表达增高有关;敲低Egr3表达可抑制冠状动脉内皮细胞炎症和凋亡.
Abstract
Objective To observe the changes of early growth response factor 3(Egr3)in coronary endothelial cells injured by high lipid and to investigate the influence of Egr3 on high lipid induced inflammation and apoptosis of coronary endothelial cells.Methods The human coronary artery endothelial cells in logarithmic growth phase were divided into control group(normal culture),high-lipid injury group(oxidized low-density lipoprotein),Egr3 no-load group(siRNA-NC lentivirus transfection+oxidized low-density lipoprotein treatment),and Egr3 knockdown group(siRNA-Egr3 lentivirus transfection+oxidized low-density lipoprotein treatment).After transfection,the relative expression of Egr3 mRNA was detected by real-time fluorescence quantitative PCR,and the relative expressions of Egr3 and caspase-3 proteins were detected by Western blot.The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the supernatant were detected by ELISA,and the apoptosis rate was detected by CCK-8.Results The relative expressions of Egr3 mRNA,Egr3 protein and caspase-3 protein,the levels of TNF-α and IL-6,and the apoptosis rate were higher in high-lipid injury group[1.56±0.48,1.65±0.35,2.43±0.43,(55.33±4.76)ng/L,(36.34± 1.60)ng/L,(186.08±20.84)％]and Egr3 no-load group[1.53±0.56,1.55±0.45,2.36±0.29,(57.85±2.92)ng/L,(31.69±1.83)ng/L,(174.34±29.98)％]than those in control group[1.04±0.20,0.51± 0.26,1.60±0.22,(46.10±1.52)ng/L,(46.10±1.52)ng/L,(23.35±1.69)ng/L,(100.01±8.64)％]and Egr3 knockdown group[0.16±0.03,0.23±0.07,1.05±0.29,(33.67±3.26)ng/L,(19.75±1.54)ng/L,(12.24± 7.51)％](P＜0.05),were higher in control group than those in Egr3 knockdown group(P＜0.05),and showed no significant differences between high-lipid injury group and Egr3 no-load group(P＞0.05).Conclusion High lipid can induce inflammation and apoptosis of coronary endothelial cells,which may be correlated with the increased expression of Egr3,and to knock down the expression of Egr3 can inhibit the inflammation and apoptosis of coronary endothelial cells.
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