红霉素对氧化应激下A549细胞的影响  

Effects of erythromycin on A549 cells under oxidative stress

作　　者：傅钰雁[1] 柳广南[1] 黄斯明[1] 郑厚文[1] 

机构地区：[1]广西医科大学第一附属医院呼吸内科,南宁530021

出　　处：《广东医学》2014年第3期340-344,共5页Guangdong Medical Journal

基　　金：广西自然科学基金资助项目(编号:2013jjAA40025)

摘　　要：目的探讨红霉素(EM)对氧化应激状态下肺腺癌细胞A549的影响。方法香烟烟雾提取物(CSE)和EM分别刺激A549细胞后显微镜下观察细胞生长情况;用MTT法检测各组细胞活性;ELISA检测各组细胞IL-8、TNF-α浓度;免疫组织化学法检测各组细胞组蛋白去乙酰化酶2(HDAC2)的表达;real-time PCR分别检测各组细胞HDAC2 mRNA、IL-8 mRNA、TNF-αmRNA的相对表达量。结果 (1)与空白对照组比较,0.5%CSE或者10μg/mL EM处理细胞48 h,细胞生长未见明显抑制;1%CSE或者20μg/mL EM处理细胞48 h,细胞生长受到显著抑制,并且在一定范围内呈剂量和时间依赖性,浓度越大,时间越长,受抑制也越严重。(2)0.5%CSE处理细胞48 h,IL-8、TNF-α均增高,而1%CSE处理细胞48 h,IL-8、TNF-α均受抑制,且随着CSE浓度增高或培养时间延长,抑制程度越严重;不同浓度EM在不同时间处理细胞,IL-8、TNF-α分泌均受到不同程度抑制。(3)0.5%CSE下调A549细胞中HDAC2的表达(P<0.001),10μg/mL EM上调氧化应激下A549细胞HDAC2的表达(P<0.05)。(4)0.5%CSE刺激细胞后,IL-8 mRNA和TNF-αmRNA表达较空白对照组均增高(P<0.05),而10μg/mL EM可降低氧化应激下细胞IL-8 mRNA和TNF-αmRNA表达(P<0.05)。结论 CSE可使A549细胞产生炎症反应增强,而EM可抑制氧化应激状态下A549细胞的炎症反应,其作用机制可能是通过上调细胞HDAC2活性而产生作用。Objective To investigate the effects of erythromycin （ EM ） on A549 cells under oxidative stress. Methods The cell growth of A549 cells exposed to different concentrations of cigarette smoke extract （CSE） and EM were observed under microscope. MTF assay was used to assess the cell viability. ELISA was used to measure IL -8 and TNF - α. Immunohistoehemistry was used to examine the expression of Histone acetylation enzyme 2 （HDAC2）. The ex- pression of HDAC2 mRNA, IL - 8 mRNA and TNF - α mRNA in A549 cells were evaluated by real - time PCR. Results Compared with control group, there was no significant inhibition on the growth of cells treated with 0. 5% CSE or 10 μg/mL EM for 48 h ; while significant inhibitions were observed in cells with 1% CSE and 20 μg/mL EM in dose - and time - dependent manners. IL-8 and TNF-α were increased in A549 cells exposed to 0. 5% CSE for 48 h, while reduced to high concentration of CSE. In addition, IL - 8 and TNF - α were suppressed by different concentrations of EM in time - dependent manner. The expression of HDAC2 was significantly down - regulated by CSE while up - regulated by EM; meanwhile, the protein and mRNA expression of HDAC2 were synchronous. The mRNA expressions of IL - 8 and TNF -α was significantly increased after 0. 5% CSE stimulating （P 〈 0.05 ）, which was reversed by 10 μg/mL EM -inducde oxidative stress （P 〈 0. 05）. Conclusion CSE enhances the inflammation of A549 cells, which can be inhibited by EM, probably via up - regulating of HDAC2.

关 键 词：红霉素 香烟烟雾提取物 A549细胞 组蛋白去乙酰化酶2 

分 类 号：R734.2[医药卫生—肿瘤]