肠道部分缺血再灌注损伤诱发外周血淋巴细胞凋亡

目的：观察肠道部分缺血再灌注(I/R)损伤过程中外周血淋巴细胞凋亡及其在植物血凝素(PHA)刺激下增殖能力的变化。方法：大耳白兔55只,分为肠部分I/R组、假手术对照组、正常对照组,用自行设计的血流阻断器阻断SMA血流50%,维持4h,制作肠道部分缺血再灌注损伤动物模型。采用流式细胞术(AnnexinV/PI双染法)测定外周血淋巴细胞凋亡变化,采用免疫组织化学方法观察外周血淋巴细胞caspase-3蛋白的表达,采用MTT比色法测定外周血淋巴细胞增殖能力。结果：肠道缺血期外周血淋巴细胞凋亡率明显升高,肠道再灌注期后外周血淋巴细胞凋亡比例骤然减少；外周血淋巴细胞caspase-3表达与凋亡改变呈相同趋势；肠部分I/R损伤后外周血淋巴细胞增殖能力明显降低。结论：肠道部分I/R损伤后,外周血T-淋巴细胞增殖功能明显受抑,外周血淋巴细胞凋亡变化与细胞内caspase-3的表达有关。

The apoptosis of peripheral blood lymphocytes induced by partially ischemia and reperfusion injury of intestine

Objective:To investigate the changes of apoptosis and the proliferative ability to PHA of peripheral blood lymphocytes induced by partially ischemia and reperfusion injury of intestine.Methods:Fifty five white rabbits were ran- domized into 3 groups:sham operation,ischemia and reperfusion(I/R),and normal control groups.The SMA were par- tially blocked with self designed instrument,producing 50% decrease in SMA blood flow,lasting for 4 hours.The chan- ges in the amount of apoptotic lymphocytes in peripheral blood were measured from 0 h to 3 days with flow cytometry (AnnexinV/PI stained),the changes of caspase-3 in peripheral blood lymphocytes were observed with histochemistry, the proliferative ability to PHA of peripheral blood lymphocytes was assayed with MTT colorimetry.Results:In I/R groups,the ratio of apoptotic lymphocytes was increased dramatically during intestinal ischemia stage,while the ratio of apoptotic lymphocytes was decreased sharply after reperfusion stage,the expression of caspase-3 showed the same trend as the apoptosis,and the proliferative ability to PHA of peripheral blood lymphocytes was damaged after partially ische- mia and reperfusion injury of intestine.Conclusion:The changes of apoptosis of peripheral blood lymphocytes is positively related to the expression of caspase-3;The proliferative ability of peripheral blood T lymphocytes was impaired after par- tially ischemia and reperfusion injury of intestine.