Developmental changes in the inhibition of cultured rat uterine cell proliferation by opioid peptides
Developmental changes in the inhibition of cultured rat uterine cell proliferation by opioid peptides
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DOI：
10.1046/j.1365-2184.2003.00263.x
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发表时间：
2003-06-01
影响因子：
8.5
通讯作者：
Vértes, M
中科院分区：
文献类型：
--
作者：
Környei, JL;Vértes, Z;Vértes, M
阿片肽是包括子宫在内的几个器官中细胞增殖的负调节剂。在本研究中，阿片肽对培养的大鼠子宫细胞增殖的直接抑制作用的个体发生进行了研究。无菌方法切除7、14、21、28、35和60日龄大鼠的子宫。组织块用胰蛋白酶和胶原酶有限消化法分散。将细胞在富集的Dulbecco改良的Eagle培养基（DMEM）中培养。在整个培养期间存在处理。通过在胰蛋白酶消化和台盼蓝排除后计数细胞来确定单层的细胞密度。大鼠子宫混合细胞培养物在10天内生长至汇合。平均种群倍增时间随动物年龄的增长而逐渐增加。表皮生长因子（EGF）增加了所有年龄组培养物的细胞密度。雌二醇（E-2）反应性出现在21日龄。[D-Met（2）-Pro（5）]-enkephalinamide（ENK）的作用呈双相性。ENK和[Met（5）]-脑啡肽（OGF）降低7日龄大鼠未刺激和EGF刺激培养物的细胞密度的程度相同。ENK在14日龄动物中无效。从21日龄开始，E-2或EGF刺激的增殖仅被ENK和DAMGO抑制，而30 nm DPDPE、Dynorhin-A、OGF、[Leu（5）]-脑啡肽、β-内啡肽和morphiceptin无效。ENK的半抑制浓度为0.3nm。ENK的作用可通过与纳洛酮联合治疗来预防。我们的新数据表明，两个不同阶段的阿片肽对大鼠子宫细胞增殖的抑制作用在个体发育与不敏感的间隔。
Opioid peptides are negative regulators of cell proliferation in several organs including the uterus. In the present study, the ontogeny of the direct inhibitory action of opioid peptides on the proliferation of cultured rat uterine cells was investigated. Uteri of 7, 14, 21, 28, 35 and 60-day-old rats were removed in a sterile way. Tissue blocks were dispersed by limited digestions with trypsin and collagenase. Cells were cultured in enriched Dulbecco's modified Eagle's medium (DMEM). Treatments were present during the entire culture period. Cell densities of the monolayers were determined by counting the cells following trypsinization and trypan blue exclusion. Rat uterine mixed cell cultures grew to confluence within 10 days. The average population doubling time gradually increased with the age of animals. Epidermal growth factor (EGF) increased cell densities of cultures from all age groups. The oestradiol (E-2 )-responsiveness appeared at 21 days of age. The effect of [D-Met(2) -Pro(5) ]-enkephalinamide (ENK) was biphasic. ENK and [Met(5) ]-enkephalin (OGF) decreased cell densities of both unstimulated and EGF-stimulated cultures from 7-day-old rats to the same extent. ENK failed to act in 14-day-old animals. From 21 days of age on, the E-2- or EGF-stimulated proliferation was inhibited only by ENK and DAMGO, while 30 nm DPDPE, Dynorhin-A, OGF, [Leu(5) ]-enkephalin, beta-endorphin, and morphiceptin were ineffective. The half-inhibitory concentration of ENK was 0.3 nm. The effects of ENK were prevented by concomitant treatment with naloxone. Our novel data demonstrate two different phases of the inhibitory action of opioid peptides on rat uterine cell proliferation during ontogeny with an insensitive interval in between.