几种药物对脂多糖诱导奶牛乳腺上皮细胞(BMECs)分泌炎性因子的影响

Effects of several drugs on secretion of inflammatory cytokines in LPS-induced bovine mammary epithelial cells in vitro

通过脂多糖(lipopolysaccharide,LPS)诱导奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMECs)建立体外炎症模型,探究不同药物对炎症模型细胞分泌炎性因子的抑制作用,为阐明药物抗炎机制奠定基础。本研究通过CCK-8法筛选LPS的最佳致炎剂量,确定细胞炎症模型;将氟尼辛葡甲胺、亚硫酸氢钠穿心莲内酯、苦木、莲胆作用于炎症模型细胞,以地塞米松作为阳性对照,ELISA法检测各组细胞培养液上清中炎性因子TNF-α、IL-6和IL-1β的表达变化情况。研究发现,经LPS(10mg/L)刺激BMECs 24h,炎症模型组细胞培养上清中TNF-α和IL-1β的含量显著高于空白对照组(P<0.05),IL-6的含量极显著高于空白对照组(P<0.01);氟尼辛葡甲胺(10mg/L)、亚硫酸氢钠穿心莲内酯(5mg/L)、苦木(5mg/L)、莲胆(5mg/L)在使用质量浓度下均能不同程度地抑制炎症模型组BMECs分泌TNF-α、IL-6和IL-1β,其中亚硫酸氢钠穿心莲内酯对炎症细胞具有很好的抗炎作用,为以后体内的抗炎作用研究奠定基础。

To explore inhibitory effect of different drug on secreting inflammatory cytokines via inflammation cell model and further study mechanism of anti-inflammatory drugs,an inflammation model was built in vitro by inducing bovine mammary epithelial cells(BMECs)with lipopolysaccharide.In this study,the optimum dose causing inflamed was screened by CCK-8,and the inflammatory cell model was determined.The flunixin meglumine,sodium bisulfite paniculta lactone,Picrasma quassioides,Liandan were used in experimental group,and the dexamethasone was used in the positive control,the expression of inflammatory factor TNF-α,IL-6 and IL-1βin the cell culture supernatant in each grup were detected by ELISA method.The study found that with the BMECs stimulated by LPS(including 10 mg/L)about 24 h,the content of TNF-αand IL-1βin the cell culture supernatant was higher in experimental group than blank control group(P<0.05),the content of IL-6 was significantly higher than that of blank control group(P<0.01);The flunixin meglumine(10 mg/L),sodium bisulfite paniculta lactone(5 mg/L),Picrasma quassioides(5 mg/L),Liandan(5 mg/L)in proper concentrations can inhibit the secretion of TNF-α,IL-6 and IL-1βin the BMECs in different level.In addition,sodium bisulfite paniculta lactone had a good anti-inflammatory effect,it laid a solid foundation for the study of the anti-inflammatory effects in the future.