DEVELOPMENT OF A NOVEL THERAPEUTIC DRUG FOR TREATMENT OF PRETERM DELIVERY

目的：绒毛膜羊膜炎（Chorioamnionitis， CAM）是早产的重要因素，羊水中各种炎症因子在CAM的发病和宫颈成熟中起重要作用。蛋白多糖（PG）是细胞外基质的主要成分，其抗炎作用引起了人们的广泛关注。本研究通过lps刺激培养的子宫颈成纤维细胞，研究PG对炎性细胞因子的影响。材料与方法：在广崎大学医院子宫肌瘤全切除术后采集正常人宫颈标本。该组织样本被切成小块，然后在含有10%胎牛血清的Dulbecco改良Eagle培养基中，在37℃湿度5% CO_2/95%空气中培养。在成纤维细胞生长到融合后，它们被用于最多六代的后续实验。将不加蛋白多糖（0.5 mg/ml、1 mg/ml）和LPS （1 μ g/ml）的子宫颈成纤维细胞分别加入或不加蛋白多糖（0.5 mg/ml、1 mg/ml）孵育48 h，获得中等分数。采用ELISA法检测培养基中IL-1β、IL-6、IL-8的含量。结果：在培养基中，体外培养的子宫颈成纤维细胞产生IL-1β的量增加，在添加LPS后12 h达到峰值，但随后逐渐下降。在培养细胞的培养基中加入PG以剂量依赖的方式降低IL-1β。以1 mg/ml PG和LPS孵育，与对照组相比，IL-1β被抑制58%。同一细胞产生的IL-6的量在IL-6的量中呈剂量依赖性。与1 mg/ml PG和LPS孵育，与对照组相比，IL-6在任何时候都显著降低。同一细胞在培养基中产生IL-8的量随时间线性增加，但IL-8的量随PG浓度的增加而下降。在0.5和1.0 mg/ml PG的存在下，IL-8的产量分别比对照降低了约20%和30.1%。结论：本研究结果表明，PG可能抑制早产，因为它抑制了多种炎症细胞因子，这些炎症细胞因子主要参与CAM的发生和宫颈成熟。少

Objectives: Chorioamnionitis (CAM) is an extremely important factor in preterm delivery, and various inflammatory cytokines of amnionic fluid have important roles in the onset of CAM and cervical ripening. Proteoglycan (PG) was a main component of the extracellular matrix and its anti-inflammatory effects have attracted particular attention. In this study, the effect of PG on inflammatory cytokines was investigated in LPS-stimulated cultured fibroblasts from the uterine cervix.Marerials and Methods: A specimen of normal human uterine cervix was collected after a total hysterectomy for uterine myoma at Hirosaki University Hospital. This tissue sample was cut into small pieces, which were then incubated in Dulbecco's modified Eagle medium containing 10% fetal bovine serum in a humidified atmosphere 5% CO_2/95% air at 37℃. After the fibroblasts had grown to confluency, they were used at up to sixth passage for the following experiments. Uterine cervical fibroblasts were incubated with or … More without Proteoglycan (0.5 mg/ml, 1 mg/ml) and LPS (1 μ g/ml) for 48 h, and a medium fraction was obtained. The amounts of IL-1β, IL-6 and IL-8 in the medium were assayed by the ELISA method.Results: In the medium, the amount of IL-1β produced by cultured fibroblasts from the uterine cervix increased and reached a peak 12 hours after the addition of LPS, but later this amount decreased gradually. The addition of PG to the medium of cultured cells reduced IL-1β in a dose-dependent manner. Incubated with 1 mg/ml PG and LPS, IL-1β was suppressed by 58% compared with the controls. The amount of IL-6 produced by the same cell in the amount of IL-6 in a dose-dependent manner. Incubated with 1 mg/ml PG and LPS, IL-6 was significantly decreased in comparison with the controls at all times. The amount of IL-8 produced by the same cell in the medium increased linearly with time, but IL-8 production decreased with an increase in PG concentration. In the presence of 0.5 and 1.0 mg/ml PG, IL-8 production was decreased by about 20% and 30.1% of the control level.Conclusions: The results of the current study indicate that preterm delivery may be suppressed by PG since it inhibited various inflammatory cytokines that mainly contribute to the onset of CAM and cervical ripening. Less