铁调素基因干扰对脓毒症小鼠淋巴细胞凋亡的影响

目的：探讨铁调素基因干扰对脓毒症小鼠淋巴细胞凋亡的影响。方法选择6-8周BALB/C 雄性健康小鼠10只，随机均分为实验组（铁调素基因干扰组，n=5）和对照组（对照RNA干扰腺病毒组，n=5）。采用尾静脉高动力注射的方法分别给予两组小鼠铁调素特异性shRNA腺病毒和对照RNA干扰腺病毒。13 d后，两组小鼠采用盲肠结扎穿孔（CLP）法复制脓毒症模型。CLP模型制备24 h后处死小鼠，留取全血和脾脏标本。采用流式细胞术测定脾脏T淋巴细胞亚型CD4＋和CD8＋的数量百分比、外周血和脾脏T淋巴细胞的凋亡百分比；采用Western blot法检测脾脏组织凋亡相关蛋白caspase-3和caspase-9的活化水平。结果实验组CLP模型24 h后脾脏CD4＋T细胞、CD8＋-T细胞百分比明显低于对照组（t分别=3.28、3.21， P均＜0.05）；实验组小鼠脾脏和外周血T淋巴细胞凋亡率较对照组明显增高（t分别=4.76、5.12， P均＜0.05）；实验组小鼠脾脏组织凋亡相关蛋白caspase-3和caspase-9活化水平较对照组明显增强（t分别=9.86、6.16， P均＜0.05）。结论铁调素基因干扰引起脓毒症小鼠淋巴细胞凋亡增加，铁调素可能在脓毒症的免疫调控中发挥重要作用。

Lymphocyte apoptosis during sepsis in Hepcidin gene-knockdown mice

Objective To investigate the effect of disruption of Hepcidin gene on lymphocyte apoptosis in septic mice. Methods Ten BALB/C male mice aged with 6-8 weeks were randomly divided into experimental group （the group of Hepcidin shRNA adenovirus, n=5） and control group （the group of control shRNA adenovirus, n=5）. The mice were ad-ministrated with Hepcidin shRNA adenovirus and control shRNA adenovirus by using high-pressure tail vein injection re-spectively. Sepsis was induced by CLP both in the experimental and control groups after 13 days. Twenty-four hours after performance of CLP, the mice were killed, and the blood and spleen samples were obtained. The percentages of CD4＋and CD8＋T lymphocytes in the spleen, as well as the percentages of T lymphocyte apoptosis in the spleen and peripheral blood were determined using flow cytometry analysis. Furthermore, the activity of pro-apoptotic proteins caspase-3 and caspase-9 in the splenic tissue was detected using Western blot analysis. Results The percentages of CD4＋and CD8＋T lymphocytes in the spleen significantly decreased in the experimental group after CLP compared to those in the control group （t=3.28, 3.21,P〈0.05）. T lymphocyte apoptosis in the spleen and peripheral blood of the experimental group increased significantly compared with those in the control group（t=4.76, 5.12, P〈0.05）. The activated levels of caspase-3 and caspase-9 in the spleen of the experimental group increased significantly compared with those in the control group （t=9.86, 6.16,P〈0.05）. Conclusion Knock-down of Hepcidin induced lymphocyte apoptosis in septic mice. Hepcidin may play an important role in immune modulation during sepsis.