Facchinetti F, Storchi A R, Petraglia F, Volpe A, Genazzani A R
Department of Obstetrics and Gynaecology, University of Modena, Italy.
Peptides. 1988 Sep-Oct;9(5):1089-92. doi: 10.1016/0196-9781(88)90094-0.
In order to evaluate the expression of the opioid precursor proopiomelanocortin (POMC) in the ovarian follicle, we measured 6 of its main end-products in 23 follicular fluids. We coupled high performance liquid chromatography (HPLC) to specific radioimmunoassays. Seven follicles were immature (diameter less than 9 mm), 10 were obtained from superovulated patients during an in vitro fertilization-embryo transfer program (greater than 22 mm) and six were persistent follicles, collected during the luteal phase [15-31 mm, luteinized unruptured follicles (LUF)]. Follicular fluids were extracted by mean of Sep-pak cartridges and then purified by HPLC with a reverse-phase C-18 column eluted in a linear gradient with acetonitrile/0.01 M hydrochloric acid (from 18:82 to 40:60). Fractions were tested with specific antisera for ACTH (1-39), alpha-MSH, beta-lipotropin (beta-LPH), beta-endorphin (beta-EP) and gamma-endorphin (gamma-EP) immunoreactivities. No presence of beta-LPH, beta-EP and ACTH was confirmed, while gamma-EP, alpha-MSH and des-alpha-MSH were detected for the first time in follicular fluid. In every class of follicles shorter chain peptides predominate over their longer chain precursor. Immature follicles are characterized by the highest amounts of gamma-EP, ACTH, alpha-MSH and des-alpha-MSH if compared to superovulated and LUF. On the contrary, beta-EP amount was highest after superovulation. Apart from this finding, peptide levels in superovulated patients and LUF are similar.(ABSTRACT TRUNCATED AT 250 WORDS)
为了评估阿片肽前体阿黑皮素原(POMC)在卵巢卵泡中的表达,我们在23份卵泡液中检测了其6种主要终产物。我们将高效液相色谱法(HPLC)与特异性放射免疫分析法相结合。7个卵泡不成熟(直径小于9毫米),10个卵泡取自体外受精 - 胚胎移植计划中超排卵的患者(直径大于22毫米),6个是在黄体期收集的持续性卵泡[15 - 31毫米,黄素化未破裂卵泡(LUF)]。卵泡液通过Sep - pak柱进行萃取,然后用反相C - 18柱进行HPLC纯化,以乙腈/0.01M盐酸线性梯度洗脱(从18:82至40:60)。用针对促肾上腺皮质激素(1 - 39)、α - 促黑素(α - MSH)、β - 促脂素(β - LPH)、β - 内啡肽(β - EP)和γ - 内啡肽(γ - EP)免疫反应性的特异性抗血清对各馏分进行检测。未证实存在β - LPH、β - EP和促肾上腺皮质激素,而γ - EP、α - MSH和去α - MSH首次在卵泡液中被检测到。在每类卵泡中,短链肽比其长链前体占优势。与超排卵卵泡和LUF相比,如果将未成熟卵泡的γ - EP、促肾上腺皮质激素、α - MSH和去α - MSH含量按最高量计算。相反,超排卵后β - EP含量最高。除此之外,超排卵患者和LUF中的肽水平相似。(摘要截短至250字)