Endocrinological and immunological study on regulatory mechanism of human endometrial differentiation

人子宫内膜取自因妇科疾病行子宫切除术的月经周期正常的患者。组织标本在含有10%胎牛血清、胶原酶和DNA酶的RPMI培养液中粉碎并孵育。采用差速沉淀法将腺体细胞与基质细胞分离。经黄体酮培养的基质细胞在形态上发生蜕膜化，并分泌催乳素进入培养上清。利用该系统研究了白细胞介素1(IL-1)、巨噬细胞集落刺激因子(M-CSF)、白血病抑制因子(LIF)和睾酮对子宫内膜分化的影响。用抗肽酶抗原的单抗对子宫内膜进行免疫组织化学检查。结果如下：1.IL-1抑制体外培养的子宫内膜间质细胞形态蜕膜化，并以剂量依赖方式减少子宫内膜间质细胞催乳素的分泌。培养的子宫内膜间质细胞在孕酮存在的情况下分泌M-CSF。加入孕酮后，M-CSF mRNA的表达增强。子宫内膜腺细胞也可分泌M-CSF，但不受性激素控制。LIF主要表达于子宫内膜腺细胞。睾酮以剂量依赖方式诱导培养的子宫内膜间质细胞分泌催乳素。黄体酮的加入促进了催乳素的分泌。子宫内膜间质细胞表达氨基肽酶N和中性内肽酶，而子宫内膜腺细胞表达二肽酶IV。这些肽酶抗原的表达在月经周期和妊娠过程中发生变化。

Human endometrium was obtained from the patients with normal menstrual cycles who underwent hysterectomy for the treatment of gynecological diseases. The tissue samples were minsed and incubated in RPMI medium containing 10 % fetal bovine serum, collagenase and DNase. Glandular cells were separated from stromal cells by differential sedimentation. The stromal cells cultured with progesterone decidualized morphologically and secreted prolactin into the culture supernatant. Using this system, the effects of interleukin 1 (IL-1), Macrophage colony-stimulating factor (M-CSF), leukemia inhibitory factor (LIF) and testosterone on the endometrial differentiation were studied. Furthermore, endometrium was examined immunohistologically with monoclonal antibodies against peptidase antigens. The results were as follows:1. IL-1 inhibited morphological decidualization of endometrial stromal cells in vitro, and also reduced prolactin secretion by the cultured endometrial stromal cells in a dose-dependent manner.2. Cultured endometrial stromal cells secreted M-CSF in the presence of progesterone. The expression of M-CSF mRNA was enhanced by the addition of progesterone. Endometrial glandular cells also secreted M-CSF, but it was not under the sex steroidal control.3. LIF was mainly detected on the endometrial glandualr cells.4. Testosterone induced prolactin secretion by the cultured endometrial stromal cells in a dose dependent manner. This prolactin secretion was enhanced by the addition of progesterone.5. Endometrial stromal cells expressed aminopeptidase N and neutral endopeptidase, whereas endometrial glandular cells expressed dipeptidyl peptidase IV. The expression of these peptidase antigens changed during menstrual cycle and pregnancy.