Ratka A, Hochhaus G, Wissler R N, Simpkins J W
Departments of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville 32610.
Mol Cell Endocrinol. 1991 Jul;78(3):155-62. doi: 10.1016/0303-7207(91)90118-c.
We have recently demonstrated that acute and chronic treatments with estradiol and progesterone induce changes in the responsiveness of endogenous opioid systems to painful stimulation. In the present study the neuroblastoma SH-SY5Y subclone known to contain predominantly mu opioid receptors was used as a model to characterize the gonadal steroid effect on this opioid receptor system. The function of opioid receptors was assessed by measuring prostaglandin E1 (PGE1)-induced cyclic AMP accumulation after various treatments with estradiol and progesterone. Differentiated SH-SY5Y cells respond to PGE1 with a dramatic increase in cAMP level. Morphine (MOR) inhibits by about 75% the stimulatory effect of PGE1 on cAMP. Pretreatment with 5 nM of estradiol for 6 days resulted in a significant increase of PGE1-stimulated cAMP accumulation. Exposure of cells for 48 h to estradiol in doses of 5 nM or 50 nM did not affect cell sensitivity to the PGE1 effect on cAMP. Moreover, neither dose of estradiol changed the inhibitory effect of morphine on PGE1-induced cAMP response. There was a significant increase in PGE1-stimulated cAMP accumulation after treatment with 100 nM progesterone for 1 h or 15 min and a marked elevation of cAMP levels was also measured after 15 min treatment with 10 nM progesterone. Exposure to either dose of progesterone for 8 h, 48 h or 6 days did not affect basal or PGE1-induced cAMP in neuroblastoma cells. Progesterone-treated groups responded to MOR with 56-67% inhibition of PGE1-stimulated cAMP accumulation. The potency of MOR-induced inhibition was comparable to the MOR effect in cells not treated with the steroid.(ABSTRACT TRUNCATED AT 250 WORDS)
我们最近证明，雌二醇和孕酮的急性和慢性治疗会引起内源性阿片系统对疼痛刺激反应性的变化。在本研究中，已知主要含有μ阿片受体的神经母细胞瘤SH-SY5Y亚克隆被用作模型，以表征性腺类固醇对该阿片受体系统的影响。通过测量在用雌二醇和孕酮进行各种处理后前列腺素E1（PGE1）诱导的环磷酸腺苷（cAMP）积累来评估阿片受体的功能。分化的SH-SY5Y细胞对PGE1的反应是cAMP水平急剧增加。吗啡（MOR）可将PGE1对cAMP的刺激作用抑制约75%。用5 nM雌二醇预处理6天导致PGE1刺激的cAMP积累显著增加。将细胞暴露于5 nM或50 nM剂量的雌二醇48小时，并不影响细胞对PGE1对cAMP作用的敏感性。此外，两种剂量的雌二醇均未改变吗啡对PGE1诱导的cAMP反应的抑制作用。用100 nM孕酮处理1小时或15分钟后，PGE1刺激的cAMP积累显著增加，用10 nM孕酮处理15分钟后也测量到cAMP水平显著升高。将细胞暴露于任何一种剂量的孕酮8小时、48小时或6天，均不影响神经母细胞瘤细胞中的基础cAMP或PGE1诱导的cAMP。孕酮处理组对MOR的反应是PGE1刺激的cAMP积累受到56 - 67%的抑制。MOR诱导抑制的效力与未用类固醇处理的细胞中MOR的作用相当。（摘要截短于250字）
