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目的探讨腺病毒E1A蛋白在致炎因素TNF-α作用下对人肺腺癌细胞(A549)炎症因子IL-8和ICAM-1表达的影响以及地塞米松(DXM)和N-乙酰半胱氨酸(NAC)的干预作用。方法构建稳定表达E1A蛋白的A549细胞系(E1A+组)及对照质粒转染细胞系(E1A-组)。用TNF-α刺激、DXM以及NAC干预细胞,用ELISA检测炎症因子IL-8蛋白的表达,流式细胞术检测ICAM-1的表达。结果E1A+组在10μg.L-1的TNF-α作用前后细胞IL-8蛋白浓度分别为(48.49±0.27)ng.L-1和(22 841.75±12.92)ng.L-1,明显高于E1A-组作用前的(1.67±0.07)ng.L-1和作用后的(3 576.04±3.20)ng.L-1,两组相比差异有统计学意义(P<0.01)。与TNF-α单独作用组相比,DXM和NAC预作用细胞可明显降低TNF-α诱导下IL-8的高表达,差异有统计学意义(P<0.01)。E1A+组在10μg.L-1的TNF-α作用前后细胞ICAM-1蛋白浓度(荧光强度)分别为17.12±3.32和35.12±3.19,均高于E1A-组作用前0.59±0.09和作用后29.72±3.32,两组相比差异有统计学意义(P<0.01)。与TNF-α单独作用组相比,DXM和NAC预作用细胞可明显降低TNF-α诱导的ICAM-1的表达,差异有统计学意义(P<0.01)。结论 E1A蛋白能够增加TNF-α诱导下的IL-8和ICAM-1蛋白的表达。DXM和NAC能够明显拮抗TNF-α诱导下的IL-8和ICAM-1的蛋白表达,具有较强的抗炎作用。腺病毒E1A蛋白对NAC及DXM的抗炎作用无明显拮抗作用。
Objective To investigate the effects of adenovirus E1A on the expression of inflammatory cytokines IL-8 and ICAM-1 in human lung adenocarcinoma cell (A549) under the action of pro-inflammatory cytokines TNF-α, and the effects of dexamethasone (DXM) and N-acetyl cysteamine Acid (NAC) intervention. Methods A549 cell line (E1A + group) and control plasmid transfected cell line (E1A - group) stably expressing E1A protein were constructed. The cells were treated with TNF-α, DXM and NAC. The expression of IL-8 was detected by ELISA. The expression of ICAM-1 was detected by flow cytometry. Results The concentration of IL-8 in E1A + group before and after TNF-α treatment was (48.49 ± 0.27) ng.L-1 and (22 841.75 ± 12.92) ng.L-1 respectively, which was significantly higher than that of E1A (1.67 ± 0.07) ng.L-1 before treatment and (3 576.04 ± 3.20) ng.L-1 after treatment, the difference was statistically significant (P <0.01). DXM and NAC preconditioning cells could significantly reduce the high expression of IL-8 induced by TNF-α, compared with TNF-α alone group, the difference was statistically significant (P <0.01). The ICAM-1 protein concentration (fluorescence intensity) in E1A + group before and after TNF-α treatment at 10μg.L-1 were 17.12 ± 3.32 and 35.12 ± 3.19, respectively, which were significantly higher than those before E1A-0.59 ± 0.09 and 29.72 ± 3.32, the difference between the two groups was statistically significant (P <0.01). DXM and NAC preconditioning cells could significantly decrease the expression of ICAM-1 induced by TNF-α compared with TNF-α alone group, the difference was statistically significant (P <0.01). Conclusion E1A protein can increase the expression of IL-8 and ICAM-1 protein induced by TNF-α. DXM and NAC can significantly antagonize the TNF-α-induced IL-8 and ICAM-1 protein expression, has a strong anti-inflammatory effect. The adenovirus E1A protein had no antagonistic effect on the anti-inflammatory effects of NAC and DXM.