摘要
目的了解在烧伤血清刺激下大鼠心肌细胞内是否存在磷脂酰肌醇3-激酶（Pl3K）／蛋白激酶B（Akt）与p38丝裂原活化蛋白激酶（p38MAPK，以下简称p38）信号通路的交叉对话，探讨此二通路在烧伤后心肌细胞损伤中的作用。方法建立烧伤血清刺激下的大鼠心肌细胞模型。（1）检测体积分数10％的烧伤血清刺激不同时间后，心肌细胞内磷酸化p38（p-p38）和磷酸化Akt（p-Akt）的表达水平。（2）检测在不同体积分数（5％、10％、20％）的烧伤血清或体积分数10％烧伤血清＋胰岛素（1×10^-6、1×10^-7、1×10^-8mol／L）作用下，心肌细胞内p-p38和p-Akt的表达水平，并测定细胞培养上清液中肌酸激酶（CK）的含量。（3）采用p38MAPK通路抑制剂SB203580、P13K／Akt通路抑制剂LY294002进行阻断实验，测定心肌细胞内p-p38和p-Akt的表达水平及细胞培养上清液中CK的含量。结果（1）体积分数10％烧伤血清作用1、3、6、12、24h时，心肌细胞p-p38水平分别为4．0±0．8、3．6±0．8、5．1±1.6、2．4±0．5、3．0±0．6，较作用0h时（加入血清后即刻）的水平（1．0）明显增高（P〈0．01）；而p-Akt表达水平分别为0．15±0．07、0．64±0．10、0．26±0．08、0．38±0．11、0．59±0．13，较作用0h时水平（1．00）明显降低（P〈0．01）。（2）不同浓度烧伤血清或烧伤血清＋胰岛索作用下，p-p38和p-Akt的表达水平呈相反变化趋势；心肌细胞CK的释放量随烧伤血清浓度升高而增高，胰岛素对此有明显的抑制作用（P〈0．05或P〈0．01）。（3）LY294002能够升高烧伤血清导致的低p—p38水平，抵消胰岛素的保护作用（P〈0．01）；SB203580能使烧伤血清所致的低p-Akt水平得以回升（P〈0．01），抑制烧伤血清引起的CK释放。结论烧伤血清作用下的心肌细胞存在Pl3K／Akt和p38信号通路的交叉对话，并可能对心肌细胞产生调控作�
Objective To investigate the possibility of crosstalk between phosphatidylinositol 3-kinase （Pl 3-K）/Akt pathway and p38 mitogen-activated protein kinase （p38MAPK） pathway in cardiomyocyte with challenge of burn serum,and to explore their influence on cardiomyocyte injury after burn. Methods The model of murine cardiomyocyte with stimulation of burn serum was established. （ 1 ） The level of Akt and p38 phosphorlation in cardiomyocyte were examined with stimulation of 10% burn serum before stimulation and 1,3,6,12,24 hour after stimulation. （2） The levels of Akt and p38 phosphorylation in cardiomyocyte were determined with stimulation of burn serum （ at concentration of 5% , 10% ,20% ） or 10% burn serum plus insulin（ at concentration of 1 × 10^-8,1 × 10^- 7,1 × 10^-6mol/L）. The content of creatine kinase （CK） in supcrnate was also detected. （3）Addition to the inhibitor of p38 MAPK pathway（SB203580） and PI3K/ Akt pathway （ LY294002 ） ,the level of p38 MAPK , PI3 K/Akt and the content of CK in supernate were determined. Results （1） The level of p-p38 in cardiomyocyte was 4. 0 ± 0.8,3.6 ±0.8,5.1 ±1.6,2.4±0.5,3.0 ±0.6 at 1,3,6,12,24 hour（s） after stimulation of burn serum,which was obviously higher than that immediate after stimulation （ 1.0, P 〈 0.01 ）. The level of p-Akt was 0.15 ± 0.07,0.64± 0.10,0.26 ± 0.08,0.38 ± 0.11,0.59 ± 0.13 ,which was obviously lower than that before stimulation （ 1.00, P 〈 0. 01 ）. （2） With stimulation of different concentration of burn serum or burn serum plus insulin, the level of p-Akt and p- p38 changed in the opposite directions comparatively. The content of CK increased along with increase of burn serum concentration ,but decreased obviously with treatment of insulin （ P 〈 0.05 or 0. 01 ）. （ 3 ）Low level of p38 induced by burn serum was increased after treatment of LY294002, which neutralized the protection of insulin（ P 〈 0. 01 ）. Low level of p-Akt induced by burn serum increa
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2008年第4期263-267,共5页
Chinese Journal of Burns
基金
国家自然科学基金（30772250）
关键词
烧伤
肌细胞
心脏
P38丝裂原活化蛋白激酶类
蛋白激酶B
烧伤血清
Burns
Myocytes, cardiac
B
Burn serum MAPK pathway in cardiomyocytes with challenge of burn p38 Mitogen-aetivated protein kinases
Protein kinase
