摘要
目的在静息状态下,以大鼠腹腔吞噬细胞[Ca2+]i和fMLP、PMA、LTB4诱导下的吞噬细胞[Ca2+]i为指标,推测吞噬细胞对fMLP、PMA、LTB4的反应性。方法通过密度梯度离心法分离大鼠腹腔吞噬细胞,将分离得到的吞噬细胞用Fura-2/AM(终浓度为5μmol/L)负载1h,采用细胞内离子测定仪测定[Ca2+]i的动态变化。结果静息状态下,巨噬细胞内[Ca2+]i高于中性粒细胞;在不同浓度fMLP、PMA、LTB4诱导下,[Ca2+]i变化都具有浓度依赖性:经fMLP诱导后,[Ca2+]i呈一过性增高,但持续时间较长,其[Ca2+]i缓慢下降,具有时间和浓度依赖性;经PMA诱导后,[Ca2+]i增高幅度最大,持续时间最长,观察45 min后也无下降趋势;经LTB4诱导后,[Ca2+]i呈一过性增高,持续时间较短,约1 min后恢复到静息状态浓度;在高浓度fMLP诱导后,约10 min后恢复到静息状态浓度。结论 fMLP、PMA、LTB4诱导引起大鼠腹腔吞噬细胞[Ca2+]i发生依赖性增高,持续时间各不相同。
objective To observe the influence of fMLP, PMA and LTB4 on [ Ca2^＋ ] i of peritoneal phagocytes and neutrophils cells under the resting state in rats. Methods The rat peritoneal phagocytes and neutrophils were separated by density gradient centrifugation, then immersed with Fura- 2/AM solution （final concentration： 5 i.Lmol/L）by l h and determined [ Ca2~ ]i by dual -wavelength fluorescence spectrophotometer. Results When the cells were in the resting state, [ Ca2^＋ ] i in phagocytes is higher than that in neutrophils. After the peritoneal phago- cytes and neutrophils were treated by different concentrations of fMLP, PMA, and LTB4, the change of [ Ca2^＋ ] i was presented concentration - dependence. The increase of [ Ca2 ＋ ~ i result by fMLP induction was also transient and concentration - dependent, but the degression was very slow, it tooklO min decreased to the level of resting state.The increase of [ Ca2~ ]i result by PMA induction was the highest and the longest, the obvious degression after 45 min was not observed. The increase of [ Ca2^＋ ] i result by LTB4 induction was transient, [ Ca2^＋ ] i decreased to the level of resting state in 1 min. Conclusion The [ Ca2^＋ ] i inside peritoneal phagocytes and neutrophils in rats induced by fMLP, PMA and LTB4, are concentration - dependent increased and the persistence time shows certain differ- ence.
出处
《青海医学院学报》
CAS
2013年第2期91-96,共6页
Journal of Qinghai Medical College
基金
教育部科学技术研究重点项目(210234)
