摘要
目的:探讨保护素(PDX)对脂多糖(LPS)诱导的原代肺成纤维细胞环氧合酶-2(COX-2)表达的影响及对转化生长因子-β_1(TGF-β_1)诱导其纤维化增殖转化的影响。方法:分离、纯化、鉴定得到小鼠肺成纤维细胞,用LPS诱导建立成纤维细胞体外炎症模型,用TGF-β_1诱导建立体外纤维化增殖模型。PDX分别作用于经过LPS或TGF-β_1诱导的原代肺成纤维细胞。采用细胞浓度试剂盒测定细胞增殖,采用实时定量PCR测定基因表达,采用ELISA法检测细胞上清液前列腺素E_2(PGE_2)水平,应用Western blot检测COX-2蛋白的表达。结果:采用10ng/mL TGF-β_1刺激诱导体外培养的原代肺成纤维细胞48h能建立较为合理的体外纤维化增殖模型。PDX抑制LPS诱导的原代肺成纤维细胞COX-2蛋白的表达及上清液PGE_2水平,同时可能通过剂量依赖方式抑制TGF-β_1诱导的原代肺成纤维细胞的增殖。PDX抑制TGF-β_1诱导的原代肺成纤维细胞collagen 1α_1、collagen 1α_2的合成以及α-SMA、纤连蛋白的基因表达。结论:PDX能抑制LPS诱导的小鼠原代肺成纤维细胞COX-2表达及PGE_2水平,同时能抑制TGF-β_1诱导的小鼠原代肺成纤维细胞纤维化增殖、胶原蛋白合成及向肌成纤维细胞转化。
Objective: To explore effects of protectin DX (PDX) on the expression of cyclooxygenase-2 (COX-2), fibroproliferation, collagen production and myofibroblast differentiation in mice primary lung fibroblast cells (LF). Methods: After the lung fibroblasts were isolated and purified, the acute inflammation model and the fibriproliferation model was respectively established with lipopolysaccharide (LPS) and with TGF-β1. Then primary lung fibroblast cells were incubated in DMEM medium containing LPS or TGF-β1 in the presence or absence of PDX. After incubation, cell viability was measured using cell titer. mRNA expression was measured by realtime-PCR. The cells were harvested, and COX-2 protein was analyzed by western-blot. Supernatants from lung fibroblast cells were collected and PGE2 protein expression was measured by ELISA. Results: It is a reasonable model of fibroproliferation in fibroblasts by administering 10 ng/mL TGF-β1 for 48 hours. PDX reduced LPS-induced expression of COX-2 and production of PGE2, while inhibiting TGF-β1-induced lung fibroblast cells proliferation in a concentration dependent manner. In addition, PDX reduced rat primary LF collagen production,α-smooth muscle actin (α-SMA) and fibronectin expression induced by TGF-β1. Conclusion: PDX inhibits LPSinduced expression of COX-2 and production of PGE2 in primary lung fibroblast cells, while decreasing TGF-β1- induced fibroproliferation, collagen production and differentiation in primary lung fibroblast cells.
作者
郑声星
潘静怡
谷丽君
杨静想
陈新瓯
李明
金胜威
ZHENG Shengxing;PAN Jingyi;GU Lijun;YANG Jingxiang;CHEN Xinou;LI Ming;JIN Shengwei(Department of Anesthesiology and Perioperative Medicine,the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)
出处
《温州医科大学学报》
CAS
2019年第6期391-396,共6页
Journal of Wenzhou Medical University
基金
国家自然科学基金资助项目(81870065
81570076)
