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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。
Objective To investigate the molecular mechanism of myo-tropinin-associated membrane protein (LAMPs) -induced heme oxygenase-1 (HO-1) expression in placental trophoblast cells. Methods The placental trophoblast cells cultured in vitro were treated with 0.5-5 μg / mL LAMPs for 4 to 12 hours. The mRNA and protein expressions of HO-1 and the expressions of nuclear factor-2 Nrf2). The enzyme activity of HO-1 was observed by colorimetric assay. The production of reactive oxygen species was detected by 2 ’, 7’-dichlorodihydrofluorescein diacetate (H2DCFDA). The effects of tyrosine kinase c-Src inhibitor PP1, reactive oxygen species inhibitor N-acetyl cysteine (NAC) and Nrf2 siRNA on HO-1 expression were observed. Results Mycoplasma genitalium LAMPs induced the expression of HO-1 mRNA and protein in trophoblast cells and up-regulated the activity of HO-1. At the same time, LAMPs can also induce its production of reactive oxygen species and promote Nrf2 nuclear translocation. After pretreatment with PP1 and NAC, the expression of HO-1 and the content of Nrf2 in the nucleus were significantly decreased. After transfection with Nrf2 siRNA, HO-1 expression was significantly reduced. Conclusion Mycoplasma genitalium (LAMP) induces the expression of HO-1 by trophoblast cells via the c-Src / ROS / Nrf2 pathway.