摘要
目的 探讨腺病毒介导的白细胞介素24 （interleukin-24,IL-24）基因转移对系膜细胞分泌细胞因子白细胞介素6 （interleukin-6,IL-6）、肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）、IL-6、TNF-α的影响.方法 通过逆转录聚合酶链式反应（reverse transcription polymerase chain reaction,RT-PCR）方法构建IL-24腺病毒载体,并经序列分析证实.腺病毒载体在293细胞中扩增.分组培养系膜细胞：对照组为5％ DMEM（dulbcco＇s modified eagle＇s medium）培养肾小球系膜细胞（glomerular mesangial cells,GMCs）;Ad.Ib24组为转染携带IL-24基因复制缺陷型腺病毒（Adenovirus.interleukin-24,Ad.IL-24）后用5％ DMEM培养肾小球系膜细胞,分别在24 h和48 h提取系膜细胞培养上清.用ELISA方法检测系膜细胞培养上清液中IL-6、TNF-α的表达.结果 ①对照组系膜细胞培养上清液中24 h和48 h的IL-6表达分别为（159.53±9.81） pg/ml和（125.78±4.61） pg/ml,Ad.IL-24组的系膜细胞培养上清液中24 h和48 h的IL-6表达分别为（812.56±7.21） pg/ml和（785.44±6.23） pg/ml.Ad.IL-24组的系膜细胞培养上清液中24 h和48 h的IL-6表达均明显高于对照组（P＜0.05）.②对照组系膜细胞培养上清液24 h和48 h中的TNF-α表达分别为（29.37±2.43） pg/ml和（26.35±3.04） pg/ml,Ad.IL-24组的系膜细胞培养上清液中24 h和48 h的TNF-α表达分别为（30.18±2.03） pg/ml和（28.16±2.35） pg/ml.Ad.IL-24组的系膜细胞培养上清液中24 h和48 h的TNF-α表达与对照组无统计学差异（P＞0.05）.结论 外源性IL-24基因转移可使系膜细胞分泌IL-6上调,而对系膜细胞分泌TNFα无明显影响.
Objective To explore the effect of adenovirus mediated interleukin-24 （IL-24）transfer on IL-6 and tumor necrosis factor （TNF）-α secreted by glomerular mesangial cells （GMCs）.MethodsIL-24 adenovirus carrier （Ad.IL-24） was constructed by RT-PCR,identified by sequencing,and am plified in 293 cells.GMCs were cultured and divided into control group （GMCs were cultured in 5％DMEM）,and Ad.IL-24 group （GMCs were infected with Ad.IL-24 and cultured in 5％ DMEM）.The supernatant of GMCs culture was collected at 24 h and 48 h.Levels of IL-6 and TNF-α in the cul ture supernatant were determined by ELISA.Results （1） IL-6 level in control group was （159.53 ± 9.81） pg/mL at 24 h and （125.78 ± 4.61） pg/mL at 48 h,and that in Ad.IL-24 group was （812.56 ± 7.21） pg/mL at 24 h and （785.44 ± 6.23） pg/mL at 48 h respectively.Secretion of IL-6 by GMCs in the culture supernatant was significantly increased by IL-24 gene delivery into GMCs （P＜0.05）.（2） TNF-α level in control group was （29.37 ± 2.43） pg/mL at 24 h and （26.35 ± 3.04） pg/mL at 48 h,and that in Ad.IL-24 group was （30.18 ± 2.03） pg/mL at 24 h and （28.16 ± 2.35） pg/mL at 48 h respectively.There was no significant change in secretion of TNF-α by GMCs in the culture su pernatant by IL24 gene delivery into GMCs （P＞0.05）.Conclusions IL-24 gene delivery into GMCs significantly increased the secretion of IL-6,but showed no impact on TNF-α secretion.
出处
《临床肾脏病杂志》
2014年第7期437-440,共4页
Journal Of Clinical Nephrology
基金
国家自然科学基金（NO.81070557）
