Brodt-Eppley J, Myatt L
Department of Obstetrics and Gynecology, Molecular and Cellular Physiology, and Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA.
Biol Reprod. 1998 Oct;59(4):878-83. doi: 10.1095/biolreprod59.4.878.
Prostaglandins synthesized at parturition may act via specific myometrial receptors as mediators of uterine contractions. Several isoforms of eicosanoid (prostaglandin) receptors, identified by pharmacological means, are linked to contractile (FP, EP1, EP3) or relaxatory (EP2, EP4, IP, DP) intracellular pathways. Changes in mRNA expression of the contractile FP and the relaxatory EP2 receptor were measured in myometrium throughout gestation, at parturition, and postpartum. Timed pregnant rats were killed at 0900 h on Day 16, 18, 20, 21, 21.5, or 22 (parturition) of pregnancy or one day postpartum (n = 5 animals/group). A longitudinal section of myometrium was removed, total RNA was extracted, and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for glyceraldehyde phosphate dehydrogenase (GAPDH), calponin, EP2, and FP receptor mRNA expression. Complementary DNA products were run on agarose gels and visualized, and the quantity of cDNA product was measured against a DNA mass ladder. RT-PCR product identity was confirmed by restriction enzyme cleavage. EP2 receptor mRNA expression was highest at Day 16 and declined significantly to Day 21.5 and one day postpartum (p < 0.05, Student-Newman-Keuls procedure). Expression of FP receptor mRNA was low at Day 16 of gestation and increased significantly until delivery (p < 0.05, ANOVA) at Day 22, then fell to prepartum levels at one day postpartum. Myometrial activity at parturition may change from an active quiescent to an active contractile state in concert with a decline in expression of the relaxatory EP2 receptors and up-regulation of contractile FP receptors.
分娩时合成的前列腺素可能通过特定的子宫肌层受体作为子宫收缩的介质发挥作用。通过药理学方法鉴定的几种类花生酸(前列腺素)受体亚型与收缩性(FP、EP1、EP3)或舒张性(EP2、EP4、IP、DP)细胞内信号通路相关。在整个妊娠期、分娩时和产后,测量子宫肌层中收缩性FP受体和舒张性EP2受体的mRNA表达变化。将定时妊娠大鼠在妊娠第16、18、20、21、21.5或22天(分娩)的09:00处死,或在产后一天处死(每组n = 5只动物)。取出子宫肌层的纵切片,提取总RNA,并对甘油醛-3-磷酸脱氢酶(GAPDH)、钙调蛋白、EP2和FP受体mRNA表达进行半定量逆转录-聚合酶链反应(RT-PCR)。互补DNA产物在琼脂糖凝胶上进行电泳并可视化,cDNA产物的量相对于DNA质量梯度进行测量。通过限制性内切酶切割确认RT-PCR产物的同一性。EP2受体mRNA表达在第