【摘要】 本研究旨在优化大鼠乳腺上皮细胞培养体系,建立LPS诱发的炎症反应模型,研究视黄醇对其炎症反应的调节机制。采用胶原酶与透明质酸酶联合消化分离大鼠乳腺上皮细胞。待细胞铺满整个培养瓶的90%时,(1)用不同浓度的LPS处理乳腺上皮细胞,24h收集细胞及培养上清液;(2)分为试验组(添加1μmol.L-1视黄醇)和对照组,处理24h后更换培养液,用10μg.mL-1的LPS处理细胞,分别于不同时间点收集细胞。结果表明:LPS处理大鼠乳腺上皮细胞后引起各个时间点炎性因子mRNA表达极显著升高,视黄醇能显著下调上述炎性细胞因子的表达。视黄醇预处理能减轻LPS诱发的大鼠乳腺上皮细胞的炎症损伤。更多还原

【Abstract】 Lipopolysaccharide(LPS) induced primary culture of rat mammary epithelial cells(rMEC) inflammatory model was established and the protective mechanism of retinoic acid was studied.rMEC s were separated through digestion by collagenase and hyaluronidase.When cells were grown to 90% confluence in 6-well plates,(1) cells were treated with different concentrations of LPS;(2) cells were treated with 1 μmol·L-1 retinoic acid before stimulation with 10 μg· mL-1 LPS.Cells and supernatants were collected.In this study,inflammatory models of primary culture of rMEC were successfully established.Stimulation of rMEC with LPS elicited a marked increase in mRNA expression for inflammatory mediators.Treatment of retinoic acid significantly decreased the mRNA expression of those mediators.Retinoic acid could reduce LPS-induced inflammatory reaction.These results further indicate that retinoic acid may protect mastitis from pro-inflammatory cytokine-mediated damage of mammary epithelial cells.更多还原