国家自然科学基金资助项目(30370165)和中国科学院知识创新工程重要方向项目(KSCX2-SW-201).
This work was supported by grants from The National Natural Sciences Foundation of China (30370165) and The Key Innovation Research Programs of The Chinese Academy of Sciences (KSCX2-SW-201).
采用半定量 RT-PCR、蛋白质印迹和免疫组化等方法,对大鼠妊娠过程中子宫和胎盘转化生长因子β1(TGF-β1) 的动态表达和表达定位,及体内注射超生理剂量的γ干扰素 (interferen-gamma, IFNγ) 后 TGF-β1 的表达进行检测,探讨了妊娠子宫和胎盘中 TGF-β1 的表达规律、定位及与 IFNγ的关系 . 正常妊娠各个时期大鼠子宫和胎盘中均能检测到 TGF-β1 mRNA 和蛋白质的表达 . 半定量 RT-PCR 结果表明:妊娠早期 (D1~D9) 子宫 TGF-β1 mRNA 的表达呈递增趋势,胚泡植入前 (D4) ,子宫 TGF-β1 mRNA 的表达较低,胚泡植入早期 (D6) TGF-β1 mRNA 的表达显著升高,植入后期 (D9) TGF-β1 mRNA 的表达达高峰,妊娠中期 (D15) 和妊娠晚期 (D19) TGF-β1 mRNA 的表达逐渐降低;妊娠早 (D9) 、中 (D15) 、晚期 (D19) 胎盘中 TGF-β1 mRNA 的表达呈时效递增, D15 胎盘中 TGF-β1 mRNA 的表达高于 D9 (P<0.01) , D19 胎盘中 TGF-β1 mRNA 的表达高于 D15 (P<0.01). 免疫组化结果显示,妊娠早期 TGF-β1 主要在子宫蜕膜中表达,中晚期主要在滋养层表达 . 生殖道肌肉注射超生理剂量的 IFNγ能下调 TGF-β1 的表达,且呈一定的剂量依赖关系 . 提示:在妊娠不同时期子宫和胎盘中 TGF-β1 的表达呈显著的时空动态变化, IFNγ酌对 TGF-β1 mRNA 的表达有调节作用 .
To investigate the expression change and location of transforming growth factor-beta1(TGF- β1) at uterus and placenta in the course of pregnancy and the effects of interferon gamma (IFNγ) on its expression, the expression of TGF-β1 was examined in uterus and placenta in different stages of gestation by methods of RT-PCR, Western blot, and immunohistochemistry respectively. And RT-PCR was used to analyze the expression of TGF-β1 mRNA after injecting different dose of IFNγ. Experimental results indicated that the expression of TGF-β1 mRNA and TGF-β1 protein were detected in uterus and placenta in every stage of gestation. The analysis of semie-quantitative RT-PCR that showed the expression of TGF-β1 mRNA was increasing gradually at uterus during the early stage of gestation (D1~D9). The expression of TGF-β1 in pre-implantation (D4) period was lower than that in implantation (D6) period and post-implantation (D9) period. The expression in post-implantation (D9) period is highest at uterus during gestation. With the process of gestation, the expression of TGF-β1 decreased significantly at uterus during mid-stage (D15) and late stage (D19) of gestation. The expression of TGF-β1 at placenta kept increasing in the course of pregnancy. The expression of TGF-β1 at mid-stage placenta (D15) was significantly higher (P<0.01) compared with that at early stage placenta (D9) and the expression of TGF-β1 at late-stage placenta D19) was significantly higher (P<0.01) compared with that at mid-stage placenta (D15). The analysis of immunohistochemistry indicated TGF-β1 mostly expressed at decidua during early stage of gestation and at trophoblast during mid-stage and late stage of gestation. After injecting exceeding normal concentration IFNγ by vaginal muscular, the expression of TGF-β1 was downregulated at uterus and placenta of pregnant rat.The results suggested that the expression of TGF-β1 was dynamic and spatio-temporal characteristics in uterus and placenta during gestation. IFNγ affected the expression of TGF-β1 at uterus and placenta in the course of pregnancy.
刘美玲,彭景楩,孙泉红,杨 颖,夏红飞.妊娠大鼠子宫和胎盘中 TGF-β1 的表达及 IFNγ 对其表达的影响[J].生物化学与生物物理进展,2005,32(5):413-420
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