Donoghue D J, Scanes C G
Department of Animal Sciences, Rutgers State University of New Jersey, New Brunswick 08903-0231.
Gen Comp Endocrinol. 1991 Dec;84(3):344-54. doi: 10.1016/0016-6480(91)90080-p.
These studies examined the cellular basis for the inhibitory effects of triiodothyronine (T3) on growth hormone-releasing factor (GRF)-evoked growth hormone (GH) release from chicken anterior pituitary cells in vitro. A primary monolayer culture of anterior pituitaries from 4- to 8-week-old White Leghorn cockerels was performed as previously described by this laboratory. Following a 72-hr preincubation period, cells were washed and incubated (2 hr) with either secretagogues or media alone (control). T3 (20 ng/ml) or vehicle was added to cells during both the preincubation (48-72 hr) and incubation (2 hr period. Triiodothyronine reduced (P less than 0.05) GH release (ng/ml) in response to (1) GRF; (2) the adenylyl cyclase stimulator, forskolin; (3) the cAMP analog and protein kinase A activator, 8-bromo cAMP; and (4) the phorbol ester and protein kinase C activator, phorbol 12-myristate 13-acetate. Triiodothyronine reduced (P less than 0.05) the intracellular content of GH and total GH (released GH and intracellular GH) irrespectively of whether secretagogues were also present. When GH release was expressed as a percentage of total GH [released GH/(intracellular GH + released GH)], percentage GH released in response to GRF, or the protein kinase A, protein kinase C, or calcium pathway activators was not as great in T3-treated versus non-T3-treated cells. These data indicate that T3 inhibits GRF-evoked GH release by reducing the availability of intracellular stores of GH and by also inhibiting second messenger-stimulated GH release pathways.
这些研究探讨了三碘甲状腺原氨酸(T3)对体外培养的鸡垂体前叶细胞中生长激素释放因子(GRF)诱发的生长激素(GH)释放产生抑制作用的细胞基础。按照本实验室先前描述的方法,对4至8周龄的白来航公鸡的垂体前叶进行原代单层培养。在72小时的预孵育期后,将细胞洗涤并与促分泌剂或单独的培养基(对照)一起孵育(2小时)。在预孵育(48 - 72小时)和孵育(2小时期间)都向细胞中加入T3(20 ng/ml)或溶剂。三碘甲状腺原氨酸降低了(P < 0.05)对以下物质产生反应时的GH释放(ng/ml):(1)GRF;(2)腺苷酸环化酶刺激剂福斯可林;(3)cAMP类似物和蛋白激酶A激活剂8 - 溴cAMP;以及(4)佛波酯和蛋白激酶C激活剂佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯。无论是否存在促分泌剂,三碘甲状腺原氨酸都降低了(P < 0.05)细胞内GH含量和总GH(释放的GH和细胞内GH)。当将GH释放表示为总GH的百分比[释放的GH/(细胞内GH + 释放的GH)]时,与未用T3处理的细胞相比,用T3处理的细胞中对GRF或蛋白激酶A、蛋白激酶C或钙途径激活剂产生反应而释放的GH百分比没有那么高。这些数据表明,T3通过减少细胞内GH储存的可用性以及抑制第二信使刺激的GH释放途径来抑制GRF诱发的GH释放。