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为了观察γ-氨基丁酸B型受体R1亚基(GABAB1R,GB1)在妊娠小鼠第1天至第8天(D1～D8)子宫中的表达规律,探讨γ-氨基丁酸(GABA)信号是否参与了胎盘早中期建立.应用半定量RT-PCR、免疫组织化学及Western blotting检测GB1在妊娠小鼠D1～D8子宫中的表达情况,原代分离D8.5的绒毛膜锥(EPCs)及蜕膜细胞,检测GABA及GABA B型受体激动剂baclofen、拮抗剂2-hydroxysaclofen对EPCs黏附及外延生长以及对蜕膜细胞侵袭能力的影响.D8时体内注射0.05 g/kg、0.5 g/kg及5 g/kg的GABA,于D14收集胎盘,制作石蜡切片.结果显示,GB1 mRNA及蛋白动态表达于D1～D8子宫.100μmol/L GABA及5μmol/L Baclofen促进EPCs的外延生长,抑制蜕膜细胞的侵袭,同时,20μmol/L 2-hydroxysaclofen成功逆转GABA对EPCs及蜕膜细胞的侵袭调节作用.5 g/kg的GABA可导致D14小鼠胎盘的迷路层滋养层细胞增多,母鼠血窦及胎鼠血管减少或发育不良,海绵滋养层的细胞滋养层细胞变小,糖原细胞减少或消失.结果提示,在小鼠早中期胎盘建立过程中,GABA信号促进滋养层侵袭至母体蜕膜组织,而抑制蜕膜细胞积极主动的侵袭行为,并可损害胎盘的结构.
In order to observe the expression regularity of γ-aminobutyric acid type B receptor R1 subunit (GABAB1R, GB1) in uterus from day 1 to day 8 (D1 ~ D8) in pregnant mice, The signal was involved in the early and middle placenta establishment.Using semi-quantitative RT-PCR, immunohistochemistry and Western blotting to detect the expression of GB1 in the uterus of D1 ~ D8 of pregnant mice, primary separation of chorionic cone (EPCs) And decidual cells were collected to test the effects of GABA and GABA receptor antagonist baclofen and antagonist 2-hydroxysaclofen on the adhesion and epitaxial growth of EPCs and the ability of decidual cell invasion.D8 injection of 0.05 g / kg, 0.5 g / kg and 5 g / kg of GABA, and the placenta was collected at D14 to make paraffin sections.The results showed that GB1 mRNA and protein were dynamically expressed in the uterus of D1-D8, 100μmol / L GABA and 5μmol / L Baclofen promoted the growth of EPCs, 20μmol / L 2-hydroxysaclofen reversed the GABA-mediated invasion and migration of EPCs and decidual cells. GABA at 5 g / kg increased the number of trophoblast cells in the stained placenta of D14 mice, Sinus and fetal rat blood vessels or dysplasia, sponge trophoblast cytotrophoblast fine And the number of glycogen cells decreased or disappeared.The results suggest that GABA signaling promotes the invasion of trophoblast into maternal decidua during early and mid-term placental establishment and inhibits the proactive invasion of decidual cells and may damage the placenta Structure.