脂多糖诱导奶牛乳腺上皮细胞先天性免疫反应

The innate immune responses of bovine mammary epithelial cells stimulated by lipopolysaccharide

采取荷斯坦奶牛乳腺,进行体外分离培养,并纯化细胞。用不同质量浓度(0、1、10、100mg/L)的脂多糖刺激乳腺上皮细胞,采用MTT法检测脂多糖对细胞增殖的影响,半定量PCR检测10mg/L的LPS对乳腺上皮细胞TLR4、TLR2、CD14、MD-2四个基因在不同时间(0、2、6h)mRNA表达水平的差异。结果表明,高剂量(100mg/L)的LPS对乳腺上皮细胞的增殖产生明显影响;LPS刺激乳腺上皮细胞后,导致TLR4、CD14、MD-2mRNA表达迅速升高,而TLR2mRNA弱表达。说明TLR4、CD14、MD-2参与LPS的识别,同时也说明脂多糖刺激乳腺上皮细胞后,乳腺上皮细胞能够产生先天性免疫反应。

The object of the paper was to study the innate immune responses of bovine mammary epithelial cells stimulated by lipopolysaccharide.Primary bovine mammary epithelial cells were divided from mammary tissue taken from Holstein cows and purified.Different concentration of LPS were co-cultured with BMEC.MTT assay was applied to demonstrate the cell growth process.At 0,2,6 h after co-culture,mRNA level of TLR4,TLR2,CD14,MD-2 were determined by semi-quantitative PCR.High concentration of LPS （100 mg/L） could depress the cell growth significantly.The increase of TLR4,CD14,MD-2 mRNA expression on BMEC were induced by LPS obviously,while no significant increase was detected in TLR2 mRNA expression.The results show that TLR4,CD14,MD-2 play an important role in the recognization procedure of LPS.Innate immune responses of BMEC could be stimulated by LPS.