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线粒体在脓毒症淋巴细胞凋亡中的作用

Study on mechanisms of mitochondria in lymphocyte apoptosis of sepsis

摘要:

目的 研究线粒体在脓毒症淋巴细胞凋亡中的作用机制.方法 用雌性、6~8周、C57BL/6小鼠建立盲肠结扎穿孔(CLP)的脓毒症动物模型.将实验动物随机分为两组:对照组、CLP组.分离脾淋巴细胞,采用Annexin V-FITC/PI标记,流式细胞仪检测脾淋巴细胞的凋亡变化;罗丹明123标记,流式细胞仪检测线粒体膜电位的变化;分离脾淋巴细胞线粒体和胞质组分,Western blot法检测细胞色素C在线粒体、胞质中的含量变化.结果 CLP组脾淋巴细胞的凋亡率为(17.3±2.2)%,较对照组的(3.5±0.5)%明显增高(P<0.05);CLP组线粒体膜电位无降低的细胞占(76.2±1.6)%,较对照组的(99.6±0.4)%明显减少(P<0.05);CLP组脾淋巴细胞线粒体和胞质内的细胞色素C含量与对照组相比,差异均有统计学意义(P<0.05).结论 在脓毒症时,线粒体及其信号传导通路在淋巴细胞凋亡中发挥了重要作用.

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abstracts:

Objective To investigate cell apoptosis, mitochondrial membrane potential,cytochrome C and mechanisms of mitochondria in lymphocyte apoptosis of sepsis. Methods In the research, female C57BL/6 mice whose body weight ranged from 17 to 25 grams were utilized and assigned randomly to two groups: sham operated group ( Control ), cecal ligation and puncture group (CLP). The present study was undertaken by using the mice splenic lymphocyte to investigate cell apoptosis,mitochondrial membrane potential, cytochrome C. The apoptosis alteration was evaluated by Annexin VFITC/PI double staining with flow cytometry. The alteration of mitochondrial membrane potential was investigated by Rhodamine-123 staining of cells. Cytochrome C of mitochondria and cytosol was investigated by Western blot methods. Statistical analysis was performed using SPSS 11.5 for Windows software.Experiment data was indicated with mean ± standard. Results The splenic lymphocyte apoptosis was significantly accelerated in the CLP group when compared with that in control group ( 17.3% ± 2.2% vs.3.5% ± 0.5%, P < 0.05 ). The Rhodamine-123 fluorescent intensity in splenic lymphocyte apoptosis was reduced in CLP group ( 76. 2% ± 1.6% ). Comparison between sham group ( 99.6% ± 0.4% ) and CLP group had statistical significance ( P<0.05 ). Apoptosis could induce mitochondrial cytochrome C release into cytoplasm. In the CLP group, elevation of cytochrome C in cytosol was concurrently in accordance with decline in mitochondrial cytochrome C content. Conclusion These data suggest that mitochondrium and mitochondria signal pathway play an important role in lymphocyte apoptosis of sepsis.

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作者: 李莉 [1] 马涛 [2] 胡文全 [2] 韩璐 [3] 王中 [2]
第一作者: 李莉
期刊: 《中华外科杂志》2010年48卷16期 1243-1246页 MEDLINEISTICPKUCSCD
分类号: R6
DOI: 10.3760/cma.j.issn.0529-5815.2010.16.011
发布时间: 2010-09-10
基金项目:
天津市应用基础研究计划面上项目
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