摘要
目的探讨氯化镧(LaCl3)对脂多糖(Lipopolysaccharide,LPS)诱导巨噬细胞钙信号通路激活及肿瘤坏死因子α(TNF-α)释放的影响。方法将巨噬细胞RAW 264.7随机分成四组:LPS组(培养液中含1μg/ml LPS),LaCl3+LPS组(以含2.5μmol/ml LaCl3的培养基孵育细胞一定时间后,再予以1μg/ml LPS刺激),LaCl3组(培养液中含2.5μmol/ml LaCl3)及对照组(培养液中不含LaCl3和LPS)。于不同时间点分别收集各组细胞及其培养上清待测。以Fluo-3/Am加载细胞,经上述不同分组处理一定时间后,流式细胞术观察细胞内Ca2+浓度([Ca2+]i)的变化;采用酶联免疫吸附试验(ELISA)检测上清中TNF-α水平。结果流式细胞术测定结果显示:LPS组细胞内[Ca2+]i为(336.67±26.16nmol/L),与对照组比较差异显著(P<0.01);LaCl3+LPS组(162.67±26.41nmol/L)显著低于LPS组(P<0.01);LaCl3组(29.84±6.42nmol/L)与对照组相比略低但无统计学意义(P>0.05)。LPS组细胞培养上清TNF-α含量(152.19±15.68pg/ml)与对照组相比,显著升高(P<0.01);而LaCl3+LPS组细胞培养上清中TNF-α含量为43.95±6.55 pg/ml,与LPS组相比显著下降(P<0.01)且与对照组相比无差异;LaCl3组细胞培养上清中TNF-α含量(27.84±3.73pg/ml)与LPS组相比显著下调(P<0.01),并且与对照组相比亦显著下调(P<0.05)。结论一定浓度的LaCl3可阻断LPS诱导巨噬细胞钙信号通路激活及抑制TNF-α的释放。
Objective To investigate the effects of Lanthanum chloride（LaCl3） on lipopolysaccharide（LPS）-induced activation of calcium signaling and release of tumor necrosis factor-α（TNF-α）.Methods RAW 264.7 cells were randomly divided into four groups： LPS group,LaCl3＋LPS group,LaCl3 group and control group.Intra-cellular Ca^2＋（[Ca^2＋]i） was detected using Fluo-3/acetoxymethyl ester by flow cytometry（FCM）.TNF-α level in culture supernatants was measured by ELISA.Results FCM assay showed that [Ca^2＋]i in the cells of LPS group was 336.67±26.16 nmol/L,significantly higher than that of control group（P〈0.01）;In LaCl3＋LPS group,[Ca^2＋]i＋ was 162.67±26.41nmol/L,which was significantly lower than that of LPS group（P〈0.01）;In LaCl3 group,[Ca^2＋]i（29.84±6.42nmol/L） was slightly lower than that of the control group with no statistical significance.TNF-α level in the cell culture supernatant of LPS group was 152.19±15.68pg/ml,which was significantly higher than that of control group（P0.01）;TNF-α level in LaCl3＋LPS group was 43.95±6.55pg/ml,which was significantly lower than that of LPS group（P〈0.01）,but there was no difference between control group and LaCl3＋LPS group.TNF-α level in the cell culture supernatant of LaCl3 group was 27.84±3.73pg/ml,which was significantly lower than that of LPS group（P〈0.01） and control group（P〈0.05）.Conclusion LPS induced calcium signaling activation can be blocked by a certain concentration of LaCl3.
出处
《实验与检验医学》
CAS
2010年第4期341-343,共3页
Experimental and Laboratory Medicine
基金
国家自然科学基金资助项目(30960405)
江西省自然科学基金(2007GZY1132)
江西省卫生厅科技计划(20071041)
关键词
氯化镧
脂多糖
钙离子
肿瘤坏死因子Α
Lanthanum chloride（LaCl3）
Lipopolysaccharide（LPS）
Ca2＋
tumor necrosis factor-α（TNF-α）
