【摘要】 目的探讨生殖支原体脂质相关膜蛋白(LAMPs)诱导胎盘滋养层细胞表达血红素氧合酶-1(HO-1)的分子机制。方法用0.5~5μg/m L生殖支原体LAMPs处理体外培养的胎盘滋养层细胞4~12 h,采用实时定量PCR和Western blot法分别检测HO-1 mRNA和蛋白的表达以及核因子相关因子-2(Nrf2)的核转位;比色法观察HO-1的酶活性;2’,7’-二氯二氢荧光黄二乙酸酯(H2DCFDA)检测活性氧产生。分别采用酪氨酸激酶c-Src抑制剂PP1、活性氧抑制剂N-乙酰半胱氨酸(NAC)和Nrf2 siRNA干预,观察HO-1的表达情况。结果生殖支原体LAMPs能诱导滋养层细胞HO-1 mRNA和蛋白的表达,上调其酶活性。同时,LAMPs也能诱导其产生活性氧,并促使Nrf2核转位。PP1和NAC预处理后,可明显降低HO-1的表达水平以及细胞核内Nrf2含量。采用Nrf2 siRNA转染后,HO-1的表达显著减少。结论生殖支原体LAMPs通过c-Src/ROS/Nrf2途径诱导滋养层细胞表达HO-1。更多还原

【Abstract】 Objective To investigate the mechanism of the expression of heme oxygenase-1( HO-1) in response to Mycoplasma genitalium-derived lipid-associated membrane proteins( LAMPs) in placental trophoblast cells. Methods Placental trophoblast cells were cultured in vitro and stimulated by 0. 5 ~ 5 μg / m L of LAMPs for 4 ~ 12 h,expression of HO-1 mRNA and protein,and nuclear translocation of nuclear factor erythroid-2 related factor 2( Nrf2) were detected by realtime PCR and Western blot,respectively. The enzyme activity of HO-1 was detected by colorimetric method. The intracellular formation of reactive oxygen species( ROS) was detected by using the fluorescent probe H2 DCFDA. Tyrosine kinase cSrc inhibitor PP1,N-acetyl-cysteine( NAC) and Nrf2 siRNA were used to analyse the function of ROS and Nrf2 in mediating of HO-1 expression. Results M. genitalium LAMPs enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in placental trophoblast cells in a concentration-dependent manner. In the meantime,LAMPs also induced placental trophoblast cells accumulation of ROS and nuclear translocation of Nrf2. PP1 and NAC treatment could inhibit LAMPs-induced HO-1 expression and Nrf2 nuclear translocation,and transfection of Nrf2 siRNA significantly abrogate HO-1 expression. Conclusion M. genitalium LAMPs induced placental trophoblast cells expression of HO-1 through c-Src / ROS / Nrf2 pathways.更多还原