Wilcox J N, Roberts J L
Endocrinology. 1985 Dec;117(6):2392-6. doi: 10.1210/endo-117-6-2392.
Ovariectomy and estrogen (E) or E plus progesterone treatment has previously been shown to alter both hypothalamic content and portal plasma levels of beta-endorphin. To determine if these changes were accompanied by changes in beta-endorphin synthesis, we used a RNA dot blot method to quantify proopiomelanocortin (POMC) mRNA levels in the arcuate nucleus-median eminence region of rats. Animals were bilaterally ovariectomized, implanted with Silastic capsules containing E or oil, and killed 1 or 3 days after implantation. Total nucleic acid was isolated from dissections of the arcuate-median eminence by proteinase-K/sodium dodecyl sulfate/phenol extraction, and POMC mRNA was quantified by dot blot analysis. Although 1 day of E treatment had no effect on hypothalamic POMC mRNA levels, 3 days of E treatment caused a significant reduction of approximately 40% of POMC mRNA levels relative to oil controls in two replicate experiments. These results suggest that the decreases in hypothalamic POMC peptide levels after E administration reported previously may be due to a decrease in POMC peptide biosynthesis resulting from a decrease in hypothalamic POMC mRNA.
卵巢切除术以及雌激素(E)或雌激素加孕激素治疗先前已被证明会改变下丘脑β-内啡肽的含量和门脉血浆水平。为了确定这些变化是否伴随着β-内啡肽合成的改变,我们使用RNA斑点印迹法来定量大鼠弓状核-正中隆起区域中阿片促黑素皮质素原(POMC)mRNA的水平。动物进行双侧卵巢切除,植入含雌激素或油的硅橡胶胶囊,并在植入后1天或3天处死。通过蛋白酶K/十二烷基硫酸钠/苯酚提取法从弓状核-正中隆起的组织中分离总核酸,并通过斑点印迹分析对POMC mRNA进行定量。虽然1天的雌激素治疗对下丘脑POMC mRNA水平没有影响,但在两项重复实验中,3天的雌激素治疗导致POMC mRNA水平相对于油对照显著降低约40%。这些结果表明,先前报道的雌激素给药后下丘脑POMC肽水平的降低可能是由于下丘脑POMC mRNA减少导致POMC肽生物合成减少所致。