Uterine-associated serine protease inhibitors stimulate deoxyribonucleic acid synthesis in porcine endometrial glandular epithelial cells of pregnancy.
Uterine-associated serine protease inhibitors stimulate deoxyribonucleic acid synthesis in porcine endometrial glandular epithelial cells of pregnancy.
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DOI：
10.1095/biolreprod61.2.380
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发表时间：
1999-08
影响因子：
3.6
通讯作者：
Lokenga Badinga;F. Michel;R. Simmen
中科院分区：
文献类型：
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作者：
Lokenga Badinga;F. Michel;R. Simmen
蛋白酶抑制剂是哺乳动物子宫的主要分泌成分，被认为主要通过调节蛋白水解酶的活性来介导妊娠相关事件。在本研究中，我们使用[(3)H]胸腺嘧啶结合试验，检测了两种丝氨酸蛋白酶抑制剂，即分泌性白细胞蛋白酶抑制剂（SLPI）和子宫纤溶酶/胰蛋白酶抑制剂（UPTI）在妊娠早期（第12天）猪子宫内膜分离的腺上皮（GE）细胞原代培养物中的有丝分裂潜能。纯化猪SLPI （pSPLI）、猪UPTI （pUPTI）或重组人SLPI （rhSLPI）均具有抗胰蛋白酶活性，在10-1000-ng/ml浓度范围内检测时，血清缺失的GE细胞DNA中胸腺嘧啶的掺入量增加（p < 0.05）。针对hSLPI或pSLPI的多克隆抗体消除了SLPI的作用。与任何一种蛋白酶抑制剂相比，pSLPI和pUPTI共同添加可提高这些细胞的DNA合成水平（p < 0.05）。糖胺聚糖肝素可以增强SLPI的抗蛋白酶活性，并有增强SLPI和UPTI诱导细胞DNA合成的趋势（p = 0.08）。逆转录聚合酶链反应表明，这两种蛋白酶抑制剂的信使rna在整个妊娠期间都存在于子宫内膜中，并且在这种组织中，GE细胞比间质成纤维细胞更大程度地存在（p < 0.05）。结果表明，除了具有良好的抗蛋白酶活性外，SLPI和UPTI可能是子宫上皮的自分泌生长促进剂。这些数据提示了一种新机制，即局部产生的蛋白酶抑制剂可能调节着床期事件和胚胎-母体通讯。
Protease inhibitors are major secretory components of the mammalian uterus that are thought to mediate pregnancy-associated events primarily by regulating the activity of proteolytic enzymes. In the present study, we examined the mitogenic potentials of two serine protease inhibitors, namely secretory leukocyte protease inhibitor (SLPI) and uterine plasmin/trypsin inhibitor (UPTI) in primary cultures of glandular epithelial (GE) cells isolated from early pregnant (Day 12) pig endometrium, using the [(3)H]thymidine incorporation assay. Purified porcine SLPI (pSPLI), porcine UPTI (pUPTI), or recombinant human SLPI (rhSLPI), all of which exhibited anti-trypsin activity, increased (p < 0.05) labeled thymidine incorporation into DNA of serum-deprived GE cells when tested at a range of 10-1000-ng/ml concentrations. Polyclonal antibodies directed against either hSLPI or pSLPI abrogated the effect of SLPI. Co-addition of pSLPI and pUPTI increased DNA synthesis in these cells to a level higher (p < 0.05) than that observed with either protease inhibitor. The glycosaminoglycan heparin, which has been previously shown to increase the anti-protease activity of SLPI, exhibited a tendency (p = 0.08) to enhance SLPI and UPTI induction of cellular DNA synthesis. Reverse transcription-polymerase chain reaction indicated that the messenger RNAs for both protease inhibitors were present in the endometrium throughout pregnancy and, within this tissue, in GE cells to a greater extent (p < 0.05) than in stromal fibroblastic cells. Results demonstrate that, in addition to their well-documented anti-protease activities, SLPI and UPTI may constitute autocrine growth promotants for the uterine epithelium. These data suggest a novel mechanism whereby locally produced protease inhibitors may modulate periimplantation events and embryo-maternal communication.