G-CSF并EPO对MODS小鼠肝细胞凋亡影响

EFFECT OF GRANULOCYTE COLONY-STIMULATING FACTOR COMBINED WITH ERYTHROPOIETIN ON LIVER CELL APOPTOSIS IN MICE WITH MULTIPLE ORGAN DYSFUNCTION SYNDROME

目的探讨粒细胞集落刺激因子(G-CSF)并促红细胞生成素(EPO)对多器官功能障碍综合征(MODS)小鼠肝细胞凋亡的影响。方法取健康雄性C57BL/6小鼠125只,随机分为正常对照组(N组)、MODS组(M组)、MODS+EPO组(M+E组)、MODS+G-CSF组(M+G组)和MODS+EPO+G-CSF组(M+E+G组)5组。采用腹腔注射酵母多糖的方法制作MODS模型。建模后第11天提取小鼠的肝组织,采用Western-blot方法检测各组肝组织中Bax、Bcl-2、Caspase-3蛋白的表达水平;采用原位末端标记技术,观察细胞凋亡情况,并计算凋亡指数(AI)。结果 M+E组、M+G组、M+E+G组凋亡蛋白Bax、Caspase-3的表达明显低于M组,差异有显著性(F=120.66,q=2.96~12.83,P<0.05);M+E+G组Bax、Caspase-3蛋白的表达明显低于M+E组、M+G组,差异有显著性(q=7.39~8.87,P<0.05)。M+E组、M+G组、M+E+G组抗凋亡蛋白Bcl-2的表达明显高于M组,差异有显著性(F=113.32,q=5.38~19.92,P<0.05);M+E+G组Bcl-2蛋白的表达明显高于M+E组、M+G组,差异有显著性(q=12.65~14.55,P<0.05)。M+E组、M+G组、M+E+G组的AI明显低于M组,差异有显著性(F=73.31,q=9.84~20.86,P<0.05)。结论 G-CSF和EPO均能在一定程度上改善MODS小鼠的肝细胞凋亡,二者联用效果优于单独使用其中任一种药物。其抗凋亡机制可能是通过上调抗凋亡蛋白Bcl-2,以及下调凋亡蛋白Bax和Caspase-3的表达来实现的。

Objective To investigate the effect of granulocyte colony-stimulating factor（G-CSF）combined with erythropoietin（EPO）on liver cell apoptosis in mice with multiple organ dysfunction syndrome（MODS）. Methods A total of 125 healthy male C57 BL/6 mouse were randomly divided into normal control group（N group）,MODS group（M group）,MODS＋EPO group（M＋E group）,MODS＋G-CSF group（M＋G group）,and MODS＋EPO＋G-CSF group（M＋E＋G group）.Intraperitoneal injection of yeast polysaccharides was performed to establish a model of MODS.Liver tissue was collected on day 11 after the model was established.Western blot was used to measure the protein expression of Bax,Bcl-2,and Caspase-3 in liver tissue,and in situ end-labeling was used to observe cell apoptosis and calculate apoptotic index（AI）. Results The M＋E group,the M＋G group,and the M＋E＋G group had significantly lower protein expression of Bax and Caspase-3 than the M group（F=120.66,q=2.96-12.83,P〈0.05）;the M＋E＋G group had significantly lower protein expression of Bax and Caspase-3 than the M＋E group and the M＋G group（q=7.39-8.87,P〈0.05）.The M＋E group,the M＋G group,and the M＋E＋G group had significantly higher expression of Bcl-2 than the M group（F=113.32,q=5.38-19.92,P〈0.05）;the M＋E＋G group had significantly higher expression of Bcl-2 than the M＋E group and the M＋G group（q=12.65-14.55,P〈0.05）.The M＋E group,the M＋G group,and the M＋E＋G group had a significantly lower AI than the M group（F=73.31,q=9.84-20.86,P〈0.05）.Conclusion Both G-CSF and EPO can improve liver cell apoptosis in MODS mice,and a combination of G-CSF and EPO has a better effect than G-CSF or EPO alone.The anti-apoptosis mechanism of G-CSF and EPO may be realized by upregulating the expression of Bcl-2 and downregulating the expression of Bax and Caspase-3.