消退素D_1抑制内毒素诱导大鼠原代肺成纤维细胞环氧合酶-2及前列腺素E_2表达的研究

Resolvin D_1 reduces expressions of cyclooxygenase-2 and prostaglandin E_2 induced by lipopolysaccharide in rat primary lung fibroblasts

目的探讨消退素D1对原代肺成纤维细胞(LF)环氧合酶-2(COX-2)及前列腺素E2(PGE2)表达的影响。方法分离、纯化、鉴定得到LF,用内毒素(LPS)干预建立成纤维细胞体外急性炎症模型,并利用细胞增殖/毒性检测试剂MTT检测出成纤维细胞急性炎症模型的最适LPS浓度和药物干预时间。分别应用不同浓度(0、10、50及100 nmol/ml)的消退素D1作用于经过LPS诱导的体外培养原代LF。采用酶联免疫法(ELISA)检测细胞上清液PGE2水平,同时应用Western-blot检测原代LF COX-2蛋白的表达。结果用1 g/ml LPS干预体外培养的原代LF 6 h能建立较为合理的急性炎症模型。消退素D1能抑制LPS诱导的原代LF COX-2蛋白的表达及上清液PGE2水平,并呈剂量依赖性。结论消退素D1能抑制LPS诱导的原代LF COX-2及PGE2的表达。

Objective To explore the effects ofresolvingDt on expression ofcyclooxygenase-2 （COX-2） and prostaglandin E2 （PGE2）in rat primary lung fibroblast cells （LF）. Methods Primary lung fibroblast cells were incuba- ted with various concentrations （0.1, 1, 10 μg/ml ）of lipopolysaccharides （LPS）for different times（3 h, 6 h, 9 h）. Then primary lung fibroblast cells were incubated in DMEM medium containing LPS in the presence or absence of resolvingD1. After incubation, the supematant was collected and the amount of PGE2 was measured by ELISA. The cells were harvested, and COX-2 protein was analyzed by western-blot. Results It was a reasonable model of acute inflammation in fibroblasts by administering 1 μg/ml LPS in fibroblasts after six hours. Induction of COX-2 protein by LPS was inhibited by resolvinD 1 in a dose dependent manner. ResolvinD1 also significantly decreased LPS induced production of PGE2. Conclusions ResolvinDl can reduce expression of LPS-induced COX-2 and consequently inhibits production of PGE2 in cultured rat primary lung fibroblast cells.