脂多糖对猪肺泡巨噬细胞分泌IL-1β及TNF-α的影响

研究了脂多糖的浓度、刺激时间对猪肺泡巨噬细胞分泌IL-1β、TNF-α的影响。分别用不同浓度的脂多糖和不同的刺激时间处理猪肺泡巨噬细胞,收集细胞,提取RNA,用实时荧光定量PCR方法检测IL-1β、TNF-αmRNA表达水平;同时收集培养上清,用ELISA检测试剂盒检测IL-1β、TNF-α蛋白表达水平。结果表明,脂多糖浓度(1~1000ng/mL)对IL-1β、TNF-α影响显著(P<0.05);刺激时间(1~24h)对IL-1β、TNF-α影响显著(P<0.05),且蛋白的表达水平迟于mRNA的表达水平。脂多糖诱导猪肺泡巨噬细胞分泌IL-1β、TNF-α具有剂量、时间依赖性。

In vitro Effects of Lipopolysaccharide on the Expression of IL-1 and TNF- in Porcine Alveolar Macrophage

The effects of various concentrations of Lipopolysaccharide(LPS) and different induced times on the expression of IL-1β and TNF-α in porcine alveolar macrophage(PAM) were investigated in this paper.PAM was maintained with different concentrations of LPS and the cell cultures were sampled at different time points.Then,the levels of mRNA expression of IL-1β and TNF-α were detected by real-time fluorescent quantitative PCR,and the levels of corresponding protein expression were simultaneously determined by commercial ELISA kit.The results showed that both of the concentration of LPS(11000ng/mL) and inducing time(1-24 h)significantly affected the expressions of IL-1β and TNF-α(P<0.05).Moreover,the expression of protein was latter than that of mRNA expression.These data indicate that LPS affects the expressions of IL-1β and TNF-α in PAM in a dose-and time-dependent manner.