Fan JiangFeng, Yu SiJiu, Cui Yan, Xu Gengquan, Wang Libin, Pan Yangyang, He Honghong
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, PR China.
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, PR China.
Theriogenology. 2017 Dec;104:23-29. doi: 10.1016/j.theriogenology.2017.07.045. Epub 2017 Jul 29.
Placental function is complex and influenced by various factors; furthermore, it depends on a delicate balance between cell proliferation, cell differentiation, and cell death. Bcl-2 and Bax proteins are key apoptosis regulators and are considered to play an important role in the maintenance of both dynamic balance and integrity of many tissues. Changes in Bcl-2 and Bax expressions have been described during different developmental stages in normal human placentas. Studies furthermore indicated several pathological placental changes to be related to abnormal Bcl-2 and Bax expressions. In the present study, we investigated both expression and distribution of Bcl-2 and Bax in yak placentas. For this, we collected placentas of 35 yaks at different stages of pregnancy as well as cotyledonary villi of four postpartum yaks. Protein and mRNA expressions of both Bcl-2 and Bax were investigated via immunohistochemistry, Western blot, and real-time PCR. Immunoreactive Bcl-2 protein was mainly localized near the fetal villous trophoblast at various gestational stages and post-partum. The Positive Index (PI) of Bcl-2 protein expression significantly decreased with increasing gestational age. Early during pregnancy (≤2 months), the Bax protein was widely distributed in the fetal villous trophoblast layer, the maternal caruncular crypt epithelium, and the stroma. Subsequently, the Bax protein distribution gradually concentrated in the fetal villous trophoblast layer. The staining intensity of Bax increased from the 3rd month to the prepartum of gestation. The PI reached a minimum of 9.4 ± 2.2 in fetal chorionic villi (FCV) and 1.3 ± 0.8 in maternal caruncular crypts (MCC) of the three months group. Both Bcl-2 and Bax had maximum immunoreactivity in the fetal villous trophoblast layer of placentas collected form postpartum yaks (with PIs of 36.6 ± 5.7 and 38.2 ± 4.8, respectively). Protein and mRNA expression of Bcl-2 and Bax investigated via Western blot and real-time PCR demonstrated similar expression profiles than immunohistochemistry. These results demonstrated the dynamic expression of Bcl-2 and Bax during pregnancy and postpartum in yak placentas. The temporal and spatial expression patterns indicate that Bcl-2 and Bax may participate in physiological processes of the placenta, such as formation, maturation, and antepartum degeneration that are critical for fetal and placental development in yak.
胎盘功能复杂,受多种因素影响;此外,它依赖于细胞增殖、细胞分化和细胞死亡之间的微妙平衡。Bcl-2和Bax蛋白是关键的凋亡调节因子,被认为在维持许多组织的动态平衡和完整性方面发挥重要作用。在正常人类胎盘的不同发育阶段,已观察到Bcl-2和Bax表达的变化。此外,研究表明胎盘的几种病理变化与Bcl-2和Bax表达异常有关。在本研究中,我们调查了牦牛胎盘Bcl-2和Bax的表达及分布情况。为此,我们收集了35头处于不同妊娠阶段的牦牛胎盘以及4头产后牦牛的子叶绒毛。通过免疫组织化学、蛋白质印迹法和实时荧光定量PCR检测Bcl-2和Bax的蛋白质和mRNA表达。免疫反应性Bcl-2蛋白在不同妊娠阶段及产后主要定位于胎儿绒毛滋养层附近。Bcl-2蛋白表达的阳性指数(PI)随胎龄增加而显著降低。妊娠早期(≤2个月),Bax蛋白广泛分布于胎儿绒毛滋养层、母体肉阜隐窝上皮和基质中。随后,Bax蛋白分布逐渐集中于胎儿绒毛滋养层。从妊娠第3个月到产前,Bax的染色强度增加。在3个月组的胎儿绒毛(FCV)中PI最低,为9.4±2.2,在母体肉阜隐窝(MCC)中为1.3±0.8。在产后牦牛胎盘的胎儿绒毛滋养层中,Bcl-2和Bax的免疫反应性均最强(PI分别为36.6±5.7和38.2±4.8)。通过蛋白质印迹法和实时荧光定量PCR检测的Bcl-2和Bax的蛋白质和mRNA表达显示出与免疫组织化学相似的表达谱。这些结果证明了牦牛胎盘在妊娠和产后期间Bcl-2和Bax的动态表达。其时空表达模式表明,Bcl-2和Bax可能参与胎盘的生理过程,如形成、成熟和产前退化,这些过程对牦牛胎儿和胎盘发育至关重要。