摘要
目的探讨活性氧(ROS)和肿瘤坏死因子-α(TNF-α)在细菌脂多糖(LPS)上调小鼠胎盘血红素氧化酶-1(HO-1)基因表达中的作用。方法由3个实验组成:实验1:经腹腔注射给予受孕第16或第17天小鼠LPS(75μg/kg),LPS处理后不同时点剖杀孕鼠;实验2:经腹腔注射给予受孕第17天小鼠不同剂量LPS(1、10、75μg/kg),LPS处理后24h剖杀孕鼠;实验3:经腹腔注射给予受孕第17天小鼠LPS(75μg/kg),部分动物于LPS处理前30min分别经腹腔注射给予2-苯叔丁基硝酮(PBN,100mg/kg)、氨基胍(AG,100mg/kg)或己酮可可碱(PTX,100mg/kg),LPS处理后1.5h或24h剖杀孕鼠。RT-PCR分析胎盘TNF-α mRNA表达改变,Western blot技术检测胎盘组织HO-1和HO-2蛋白表达水平,Griffith法测定胎盘谷胱甘肽含量,比色法检测胎盘组织羰基产物丙二醛含量。结果LPS对小鼠胎盘HO-1表达有显著的上调作用,并呈明显的时间和剂量-效应关系。PBN或PTX预处理均明显减弱LPS对胎盘组织HO-1表达的上调作用,而AG对LPS上调胎盘HO-1表达的影响不明显。结论ROS和TNF-α可能至少部分参与LPS对胎盘HO-1表达的上调作用。
Objective To investigate the role of reactive oxygen species (ROS) and tumor necrosis factor alpha (TNF-α) on lipopolysaccharide (LPS) -induced upregulation of heme oxygenase (HO) -1 in mouse placenta. Methods The present study included three separate experiments. In experiment 1 ,the pregnant mice were administered with a single dose of LPS (75μg/kg,ip) on gestational day 16 or 17 and sacrificed at 2,12,24 or 48 h after LPS treatment. In experiment 2, the pregnant mice were administered with different doses of LPS (1,10,75μg/kg,ip) on gestational day 17 and sacrificed at 24 h after LPS treatment. In experiment 3, the pregnant mice were pretreated with either PBN ( 100 mg/kg,ip) or AG ( 100 mg/kg,ip) or PTX ( 100 mg/kg,ip) at 30 min before LPS and sacrificed at 1.5 h or 24 h after LPS administration. TNF-α mRNA in mouse placentas was measured by RT-PCR. The expressions of HO-1 and HO-2 in mouse placentas were determined using Western blot. GSH was determined by the method of Griffith and lipid peroxidation was quantified by measuring malondialdehyde (MDA). Results LPS sig nificantly up-regulated the expression of HO-1 in mouse placenta inoa time- and dose-dependent manner. Either PBN or PTX pretreatment significantly attenuated LPS-induced up-regulation of placental HO-1 ,whereas AG had little effect on LPS-induced up-regulation of HO-1 in mouse placenta. Conclusion LPS-induced up-regulation of placental HO-1 is probably mediated,at least in part,by ROS and TNF-α.
出处
《安徽医科大学学报》
CAS
北大核心
2008年第4期362-366,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金项目(编号:30572223
30671786)
关键词
脂多糖类
胎盘
活性氧
肿瘤坏死因子Α
lipopolysaccharides
placenta
reactive oxygen species
tumor necrosis factor-alpha