研究μ阿片受体对卵母细胞早期发育成熟过程中的影响

Opioid receptors leads to luteinizing hormone, follicle-stimulating hormone concentrations lower direct impact on ovarian function, resulting in target organ dysfunction uterus. It has also been reported that μ opioid receptor expressed in human oocytes. So with increasing drug and prolonged anesthesia time, μ opioid receptor whether accumulation in oocytes, wound affect oocyte meiotic and early embryonic development? These problem had no related literature reports at present. This paper intends to use opioid receptor agonist - Fentanyl take the oocytes; opioid receptor antagonist - naloxone collaborate anesthesia the oocytes retrieval. Using RT-qPCR, Western-blot and the expression of various developmental stages immunofluorescence oocytes μ opioid receptor, the use of confocal microscopy process of oocyte development in chromosome alignment and separation of homologous chromosomes. These explore the μ-opioid receptor and oocyte maturation and early development whether a certain relationship.

阿片受体导致黄体生成素、卵泡刺激素的浓度降低，直接影响卵巢功能，造成靶器官子宫的功能紊乱。也有报道，μ阿片受体在人类卵母细胞中表达。那么随着麻醉时间的延长和药量的增加，μ阿片受体是否会在卵母细胞中蓄积，是否会影响卵母细胞减数分裂及早期胚胎发育？这些问题目前没有相关文献报道。本课题拟运用阿片受体激动剂——芬太尼麻醉取卵；应用阿片受体拮抗剂——纳洛酮进行协同麻醉取卵。运用RT-qPCR，Western-blot和免疫荧光技术检测卵母细胞各发育期μ阿片受体的表达变化，运用共聚焦显微镜观察卵母细胞发育过程中对染色体排列和同源染色体分离过程。从而探讨μ阿片受体与卵母细胞成熟和早期发育是否有一定的关系。

本课题拟运用阿片受体激动剂——芬太尼麻醉取卵；应用阿片受体拮抗剂——纳洛酮进行协同麻醉取卵。运用免疫荧光技术和Western-blot技术检测卵母细胞各发育期μ阿片受体的表达变化，运用共聚焦显微镜观察卵母细胞发育过程中对染色体排列和同源染色体分离过程。结果发现，高浓度的芬太尼（5μg/ml）抑制细胞染色体分裂，而浓度为1.5μg/ml的芬太尼不对细胞生长分裂产生影响，但低浓度芬太尼（0.5μg/ml）促进细胞染色体提前分裂。选择浓度为1.5μg/ml的芬太尼干预的卵母细胞，加入纳洛酮拮抗，发现高浓度的纳洛酮（32.7mg/L）抑制卵母细胞生长发育，而浓度为0.327 mg/L的纳洛酮对1.5μg/ml芬太尼干预的卵母细胞不影响生长发育，同时低浓度纳洛酮（3.27×10-3 mg/L）具有促进细胞染色体提前分裂的作用。Western-blot技术检测1.5μg/ml，芬太尼干预的卵母细胞，卵母细胞从GV期到MⅡ期卵母细胞中的μ 阿片受体均有表达，且各期μ 阿片受体的表达量无明显差异。