摘要
目的探讨p38丝裂素活化蛋白激酶（MAPK）信号转导通路在脂多糖（LPS）作用下血管平滑肌细胞（VSMc）分泌白细胞介素-6（IL-6）中的调节作用。方法将体外培养的大鼠胸主动脉VSMC分为LPS刺激组、p38MAPK抑制剂SB203580干预组、SB203580对照组和溶液对照组。LPS组以终浓度100μg／L的LPS与VSMc共同孵育；干预组VsMC以p38MAPK抑制剂SB20358010μmol／L预处理2h，再加入终浓度100pg／L的LPS共同孵育；对照组仅以SB20358010μmol／L预处理2h；溶液组仅加入去血清培养液培养。各组于培养0、3、6、12、24h后采用实时聚合酶链反应（real—timePCR）和酶联免疫吸附法（EuSA）分别检测细胞IL-6mRNA和上清液中IL-6蛋白表达。结果LPS刺激3h，VSMC中IL-6mRNA和蛋白表达即出现明显增高[mRNA（21．3±3．2）×104，蛋白（296．2±19．6）ng／L]，12h达高峰[mRNA（131．4±11．2）×104，蛋白（897．7±34．0）ng／L]，24h有所降低[mRNA（15．3±4．7）×104，蛋白（194．3±24．0）ng／L]，但仍显著高于溶液组（mRNA（9．4土1．9）×104，蛋白（29．4土4．4）ng／L，均P〈0．053。干预组3、6、12h可明显抑制LPS诱导VSMC中IL-6的分泌[mRNA（15．4±3．6）×104、（43．2土6．6）×104、（56．2±5．5）×104，蛋白（180．3土23．6）、（432．2土56．8）、（546．2士57．9）ng／L，均P〈0．053。结论LPS诱导VsMc可明显增加IL-6的mRNA和蛋白表达，p38MAPK抑制剂SB203580可显著抑制IL-6转录和蛋白合成，表明p38MAPK信号转导通路可能通过直接或间接作用参与了IL-6的分泌调节作用。
Objective To investigate the regulation mechanism of p38 mitogen-activated protein kinase （p38MAPK） in interleukin-6 （IL-6） expression of vascular smooth muscle cell （VSMC） induced by lipopolysaccharide （LPS）. Methods Rat VSMCs were divided into LPS group, SB203580＋LPS group, SB203580 group and control group. LPS group was treated with 100 tlg/L LPS for 0, 3, 6, 12, 24 hours, SB203580＋LPS group was first treated with 10μmol/L SB203580 for 2 hours and then exposed to 100 μg/L LPS for 0, 3, 6, 12, 24 hours, SB203580 group was pretreated with 10 μmol/L SB203580 for 2 hours. The level of IL-6 mRNA was determined by real-time polymerase chain reaction （PCR） and IL-6 secretion in the culture medium was measured by enzyme linked immunosorbent assay （ELISA） at different time points. Results The expression of IL-6 mRNA and the release of IL-6 were increased significantly in VSMC as early as 3 hours after being treated with LPS CmRNA： （21.3± 3.2） ）〈 104, protein.. （296.25± 19.6） ng/L], peaked in 12 hours [mRNA： （131.4 ±11.2） × 104, protein： （897.7 ± 34.0） ng/L], and the elevation persisted up to 24 hours after treatment [mRNA： （15.3 5= 4.7） × 104, protein： （194. 3 ± 24.0） ng/L] compared with control group [mRNA： （9.4±1.9））〈 104, protein.. （29.4±4. 4） ng/L, all P〈0. 053. On the other hand, the expression of IL-6 was significantly suppressed by p38MAPK inhibitor SB203580 at 3, 6, 12 hours [mRNA ： （15.4 ± 3.6） ）〈 104, （43.2 ± 6.6） ）〈 104, （56.2 ± 5.5） × 104, protein ： （180.3 ×23.6）, （432.2×56.8）, （546.2×57.9） ng/L, all P〈0. 053. Conclusion The release of IL-6 and the expression of IL-6 mRNA was increased significantly in LPS-challenged VSMC; however, the induction of IL-6 was significantly suppressed by p38MAPK inhibitor, p38MAPK may play an important role in the release of IL-6 induced by LPS.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2010年第5期291-294,共4页
Chinese Critical Care Medicine
基金
山东省医药卫生科技发展计划项目（2009HZ055）
作者简介
通信作者：王春亭，Email：wangchunting7051@126．com
