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目的探讨活性氧(ROS)在细菌脂多糖(LPS)上调小鼠胎肝血红素氧化酶-1(HO-1)基因表达中的作用。方法本研究由3个部分组成:一、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;二、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24 h剖杀孕鼠;三、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30 min分别经腹腔注射给予2-苯叔丁基硝酮(PBN,100 mg/kg)、氨基胍(AG,100 mg/kg)或己酮可可碱(PTX,100 mg/kg),LPS处理后1.56、或24 h剖杀孕鼠。用免疫测定法(ELISA)技术检测母鼠血清和羊水中TNF-α水平,用Griess反应法分析母鼠血清和羊水中一氧化氮(NO)的产生,Western blotting技术检测胎肝组织HO-1和HO-2蛋白表达水平,Griffith法测定胎肝还原型谷胱甘肽(GSH)含量,比色法检测胎肝组织羰基产物丙二醛(MDA)含量。结果LPS对小鼠胎肝HO-1表达有显著的上调作用,并呈明显的时间-效应关系。PBN预处理阻断LPS对胎肝组织HO-1表达的上调作用,而AG和PTX对LPS上调胎肝HO-1表达的影响不明显。结论ROS参与LPS对胎肝HO-1表达的上调作用。
Objective To investigate the role of reactive oxygen species (ROS) in the up-regulation of heme oxygenase-1 (HO-1) gene expression in fetal liver induced by bacterial lipopolysaccharide (LPS). Methods The study consisted of three parts: one, intraperitoneal injection of LPS (75μg / kg) on the 16th or 17th day of pregnancy, and the pregnant rats were dissected at different time points after LPS treatment. Second, LPS (1, 10, 75μg / kg) at different doses for 17 days and 24 hours after LPS treatment. Third, mice were given LPS (75μg / kg) by intraperitoneal injection on the 17th day, The animals were anesthetized by intraperitoneal injection of 2-phenylbutyl nitrone (PBN, 100 mg / kg), aminoguanidine (AG, 100 mg / kg) or pentoxifylline (PTX, 100 mg / kg) After LPS treatment 1.56, or 24 h cut off pregnant rats. The levels of TNF-αin serum and amniotic fluid of maternal rats were detected by immunoassay (ELISA), and the production of nitric oxide (NO) in serum and amniotic fluid of female rats was analyzed by Griess reaction. HO-1 And HO-2 protein expression levels were measured. Griffith’s method was used to determine the contents of GSH in fetal liver and the malondialdehyde (MDA) content in fetal liver tissue by colorimetry. Results LPS had a significant up-regulation of HO-1 expression in fetal liver and showed a time-effect relationship. Pretreatment with PBN blocked the up-regulation of HO-1 expression in fetal liver tissue by LPS, while AG and PTX had no obvious effect on LPS up-regulated HO-1 expression in fetal liver. Conclusion ROS upregulates the HO-1 expression in fetal liver induced by LPS.