Abstract:
The current study aimed to explore the molecular mechanism by which the cholecystokinin (CCK)-mediated CCKAR and CCKBR, as well as the molecular mechanisms of CCK-mediated insulin signalling pathway, regulate oestrogen in the granulosa cells. Also, the expression of CCK in ovaries, uterus, hypothalamus and pituitary gland was investigated in Camelus bactrianus. Ovaries, uterus, hypothalamus and pituitary gland were collected from six, three before ovulation (control) and three after ovulation, slaughtered Camelus bactrianus. Ovulation was induced by IM injection of seminal plasma before slaughtering in the ovulated group. The results showed that there were differences in the transcription and protein levels of CCK in various tissues before and after ovulation (p < .05, p < .01). After transfection with p-IRES2-EGFP-CCK, the mRNA and protein levels of CCK, CCKAR, CCKBR and ER in follicular granulosa cells were significantly upregulated (p < .05, p < .01), and the content of E2 was significantly upregulated (p < .01); On the contrary, after transfection with si-CCK, the mRNA and protein levels of CCK, CCKAR, CCKBR and ER in follicular granulosa cells were significantly downregulated (p < .05, p < .01), and the content of E2 was significantly downregulated (p < .01). Regulating CCK can affect the mRNA levels of INS, INSR, IGF and IGF-R. In summary, regulating the expression level of CCK can activate insulin-related signalling pathways by CCKR, thereby regulating the steroidogenic activity of granulosa cells.
摘要:
本研究旨在探讨胆囊收缩素(CCK)介导CCKAR和CCKBR的分子机制,以及CCK介导的胰岛素信号通路的分子机制,调节颗粒细胞中的雌激素。此外,CCK在卵巢中的表达,子宫,在双峰驼中研究了下丘脑和垂体。卵巢,子宫,下丘脑和垂体是从六个收集的,排卵前三个(对照)和排卵后三个,宰杀的双峰驼。排卵组在屠宰前通过IM注射精浆诱导排卵。结果表明,CCK在排卵前后各组织中的转录水平和蛋白水平存在差异(p<.05,p<.01)。用p-IRES2-EGFP-CCK转染后,CCK的mRNA和蛋白质水平,CCKAR,卵泡颗粒细胞中的CCKBR和ER显著上调(p<.05,p<.01),E2的含量显著上调(p<0.01);相反,用si-CCK转染后,CCK的mRNA和蛋白质水平,CCKAR,滤泡颗粒细胞中的CCKBR和ER显著下调(p<.05,p<.01),E2含量显著下调(p<0.01)。调节CCK可以影响INS的mRNA水平,INSR,IGF和IGF-R总之,调节CCK的表达水平可以通过CCKR激活胰岛素相关的信号通路,从而调节颗粒细胞的类固醇生成活性。