DEVELOPMENT OF A NOVEL THERAPEUTIC DRUG FOR TREATMENT OF PRETERM DELIVERY

目的：绒毛膜羊膜炎(Chorioamnionitis，CAM)是导致早产的重要因素，羊水中的各种炎性细胞因子在CAM的发生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，其抗炎作用备受关注。在本研究中，我们观察了PG对培养的子宫颈成纤维细胞炎性细胞因子的影响。材料和方法：收集广崎大学附属医院子宫肌瘤全子宫切除术后的人正常宫颈标本。将组织标本切成小块，加入含10%胎牛血清的Dulbecco改良EAGLE培养液中，37℃，湿度为5%CO_2/95%空气。在成纤维细胞生长到融合后，它们被用于随后的实验，直到第六代。宫颈成纤维细胞与或…共同孵育不含蛋白多糖(0.5 mg/m l、1 m g/m l)和内毒素(1μg/m l)作用4 8h，得到中等级分。结果：培养的宫颈成纤维细胞分泌IL-1β、IL-6、IL-8的量在培养的宫颈成纤维细胞中增加，12h达高峰，以后逐渐下降。在培养细胞的培养液中加入PG可剂量依赖性地降低IL-1β。1 mg/mlPG和内毒素孵育后，IL-1β的抑制率为58%。同一细胞产生的IL-6量与IL-6量呈剂量依赖关系。1 mg/mlPG和内毒素孵育后，IL-6在各时间点均显著低于对照组。同一细胞在培养液中产生IL-8的量随时间呈线性增加，而IL-8的产生则随PG浓度的增加而减少。在0.5和1.0 mg/mlPG的存在下，IL-8的产生分别比对照组减少20%和30.1%。结论：PG可能通过抑制多种炎性细胞因子而抑制早产，这些细胞因子是导致CAM和宫颈成熟的主要因素。较少

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