Kisielewska J, Zezula-Szpyra A, Bilińska B, Flint A P, Ziecik A J
Division of Reproductive Endocrinology and Pathophysiology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn.
Folia Histochem Cytobiol. 1997;35(3):143-9.
The objective of presented studies was to investigate whether estradiol and progesterone administered in vivo and/or added in vitro can influence the primary myometrial cell culture and how these steroid hormones can affect hCG stimulated cAMP and inositol phosphate production in the porcine uterine myocytes. Myometrial smooth muscle cells were obtained from six ovariectomized gilts pretreated (n = 4) or not (n = 2) with oestradiol benzoate and progesterone for 5 consecutive days. Immunocytochemical staining proved that the pattern of filamentous actin in the cytoplasm of the myometrial fibroblasts (basketlike network) was different from that of myometrial smooth muscle cells (long parallel fibres). The myocytes derived from steroid treated pigs and supplemented with estradiol and progesterone in vitro formed a hillock pattern on days 4-5 day of culture whereas cells obtained from not steroid pretreated gilts were smaller and did not create confluent form. Myocytes were treated in vitro with two doses of estradiol/progesterone (low - 0.2 nM/50 nM and high - 2 nM/500 nM, respectively) and two doses of hCG - 0.1 mU and 1000 mU/ml to study hCG action on the second messenger system in myocytes. The myometrial smooth muscle cells treated with low dose of estradiol and progesterone in vivo responded with much higher accumulation of inositol phosphates to strong (1000 mU/ml) hCG stimulation when compared with those receiving high dose of both steroids. The different doses of estradiol and progesterone caused a similar increase in basal cAMP accumulation as compared to control cells cultured without steroid hormones. hCG (0.1 mU/ml) had usually the additive effect on cAMP production in porcine myometrial cells. The presented paper shows that estradiol and progesterone administration in vivo followed by steroid hormone treatment in vitro affects the primary myometrial cells culture and that both steroid hormones modify the basal accumulation of the second messengers: cAMP and IP3 and their answer to hCG stimuli in pigs.
本研究的目的是调查体内给予和/或体外添加雌二醇和孕酮是否会影响原代子宫肌层细胞培养，以及这些甾体激素如何影响猪子宫肌细胞中hCG刺激的cAMP和肌醇磷酸的产生。从六只卵巢切除的小母猪获取子宫肌层平滑肌细胞，其中四只连续5天用苯甲酸雌二醇和孕酮预处理，另外两只未进行预处理。免疫细胞化学染色证明，子宫肌层成纤维细胞（篮状网络）细胞质中丝状肌动蛋白的模式与子宫肌层平滑肌细胞（长平行纤维）不同。来自经甾体处理的猪并在体外补充雌二醇和孕酮的肌细胞在培养的第4 - 5天形成小丘模式，而从未经甾体预处理的小母猪获得的细胞较小，未形成汇合形态。体外使用两剂量的雌二醇/孕酮（低剂量分别为0.2 nM/50 nM和高剂量分别为2 nM/500 nM）和两剂量的hCG - 0.1 mU和1000 mU/ml处理肌细胞，以研究hCG对肌细胞中第二信使系统的作用。与接受高剂量两种甾体激素的细胞相比，体内用低剂量雌二醇和孕酮处理的子宫肌层平滑肌细胞对强（1000 mU/ml）hCG刺激的肌醇磷酸积累反应更高。与在无甾体激素条件下培养的对照细胞相比，不同剂量的雌二醇和孕酮导致基础cAMP积累有相似增加。hCG（0.1 mU/ml）通常对猪子宫肌层细胞中cAMP的产生有相加作用。本文表明，体内给予雌二醇和孕酮后再进行体外甾体激素处理会影响原代子宫肌层细胞培养，并且两种甾体激素都会改变第二信使cAMP和IP3的基础积累以及它们对猪hCG刺激的反应。
