摘要
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.
To explore the expression of GB1 in mouse uterus from day 1 to day 8 of pregnancy （D1 -D8） and the role in placenta establishment, semi-quantitative reverse transcription polymerase chain reaction （semi-qRT-PCR）, immunohistochemistry and Western blotting were applied to detect the expression levels of GB 1 mRNA and protein respectively from D1 to D8. Besides, EPCs and decidual cell were dissected out from D8.5 uterus and then added different concentrations of GABA, GABA B receptor agonists baclofen and antagonists 2-hydroxysaclofen. In vitro attachment and outgrowth assays were performed in EPCs, and transwell chambeer was performed in decidual cells. In vivo, mouse from D8 to D13 were treated by intraperitoneal injection of different concentrations of GABA. On D14, the placenta structure was examined with the histological method. The results suggested that GB1 mRNA and protein dynamic expressed from D1 to D8 uterus. 100 μmol/L GABA and 5 μmol/L baclofen promoted EPCs outgrowth and surppressed decidual cells invasion. At the same time, 20 μmol/L 2-hydroxysaclofen could reversed both of the functions of GABA on EPCs and decidual cells. The histological structures of placentas changed appearantly on D 14. In the layer of labrinthine, the cells arranged crowdly and maternal blood and fetal rat vascular decreased number or dysplasia. In spongiotrophoblast, cytotrophoblast cells becomed smaller and glycogen cells decreased or disappeared. The combined results indicated during the early-middle mouse placenta establishment GABA signal was in favour of trophoblasts invasion but played converse role in decidual cells and damaged placenta structure.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2012年第12期1207-1214,共8页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金面上项目(30770816
31171436)
重庆市教委科学技术研究(KJ110316)资助项目~~
