Yuan W, Greenwald G S
Department of Physiology, Ralph L. Smith Research Center, University of Kansas Medical Center, Kansas City 66160-7401.
Biol Reprod. 1994 Sep;51(3):472-9. doi: 10.1095/biolreprod51.3.472.
We have recently shown that FSH, LH, and prolactin (PRL)--alone or combined--act as luteotropins when incubated with luteal cells from pregnant hamsters (Yuan and Greenwald, Biol Reprod 1994; 51:43-49). The purpose of the present study was to determine which second messenger systems are affected by these hormones with progesterone (P4) synthesis as the principal endpoint after 4 h of incubation with 100,000 luteal cells. Luteal cells on Days 4, 10, or 12 of pregnancy were incubated with the following reagents: 10 ng of recombinant human FSH (r-hFSH), ovine (o) FSH, oLH, oPRL, forskolin, db-cAMP, protein kinase A inhibitor (PKI), protein kinase C activator (phorbol 12-myristate 13-acetate; PMA), or various combinations of the reagents. Forskolin and db-cAMP each stimulated P4 in a dose-dependent manner, while PKI significantly inhibited forskolin-, r-hFSH-, oFSH-, and oLH-stimulated P4 on Day 4 of pregnancy. PMA (0.001-1.0 microM) did not affect basal P4 on Day 4, 10, or 12 of pregnancy; however, 100 nM PMA inhibited db-cAMP-, forskolin-, oFSH-, and oLH-stimulated P4 synthesis on Days 4 and 12. The antagonistic effects of PMA were reversed in all cases by concurrent incubation with a PKC inhibitor, H-7. On Day 4 of pregnancy, P4 was stimulated by oFSH and oLH with the highest levels observed in medium stimulated by the luteotropic complex of oFSH, oLH, and oPRL. Recombinant hFSH enhanced P4 production in a dose-dependent manner; doses of 10 ng and above resulted in statistically significant differences from the control values (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
我们最近发现,当与妊娠仓鼠的黄体细胞共同孵育时,促卵泡激素(FSH)、促黄体生成素(LH)和催乳素(PRL)单独或联合作用时可作为促黄体素(Yuan和Greenwald,《生物繁殖》1994年;51:43 - 49)。本研究的目的是确定在与100,000个黄体细胞孵育4小时后,以孕酮(P4)合成作为主要终点时,这些激素会影响哪些第二信使系统。将妊娠第4、10或12天的黄体细胞与以下试剂共同孵育:10 ng重组人FSH(r - hFSH)、羊(o)FSH、oLH、oPRL、福斯高林、二丁酰环磷腺苷(db - cAMP)、蛋白激酶A抑制剂(PKI)、蛋白激酶C激活剂(佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯;PMA)或这些试剂的各种组合。福斯高林和db - cAMP均以剂量依赖性方式刺激P4生成,而PKI在妊娠第4天显著抑制福斯高林、r - hFSH、oFSH和oLH刺激的P4生成。PMA(0.001 - 1.0 microM)在妊娠第4、10或12天不影响基础P4;然而,100 nM PMA在第4天和第12天抑制db - cAMP、福斯高林、oFSH和oLH刺激下的P4合成。在所有情况下,通过与PKC抑制剂H - 7共同孵育可逆转PMA的拮抗作用。在妊娠第4天,oFSH和oLH刺激P4生成,在由oFSH、oLH和oPRL的促黄体复合物刺激的培养基中观察到最高水平。重组hFSH以剂量依赖性方式增强P4生成;10 ng及以上剂量与对照值相比有统计学显著差异(p < 0.05)。(摘要截断于250字)