The Study Of The Effect Of Hepcidin Knock-Down On Lymphocyte Apoptosis In Septic Mice

Objective:To investigate the effect of hepcidin on lymphocyte apoptosis in septic mice, and provide a theoretical reference for the pathogenesis and treatment of sepsis.Methods:Ten BALB/C male mice aged with6-8weeks were randomly divided into the experimental group (the group of hepcidin-knockdown, n=5) and the control group (the group of siRNA adenovirus, n=5). The experimental group was administrated with hepcidin specific siRNA adenovirus through tail vein by high-pressure injection. The control mice were given with control siRNA adenovirus using the same method. Mice were free for diet and activity. The sepsis model was induced by CLP both in the experimental and control groups after13days. The mice were killed after24hours, and the blood and spleen were obtained. The percentage of CD4+and CD8+T lymphocytes subgroup in the spleen were determined using flow cytometry analysis. The apoptosis of spleen cells was detected by means of TUNEL. The percentage of T lymphocyte apoptosis in the spleen and peripheral blood were detected through Annexin-V flow cytometry analysis. The activity of caspase-3and caspase-9in splenic tissue was detected using Western blot analysis.Result: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 (CD4+:17.34%vs.29.08%, P<0.05; CD8+:8.37%vs.14.57%, P<0.05). Apoptosis in the spleen tissue of the experimental mice was more obvious than that in the control group by means of TUNEL. T lymphocyte apoptosis in the spleen and peripheral blood of the experimental group increased significantly compared with those in the control group (spleen:79.3%vs.59.6%, P<0.05; peripheral blood:93.0%vs.68.9%, P<0.05). The activated levels of caspase-3and caspase-9in the spleen of the experimental group increased significantly compared with those in the control group (caspase-3:11.97vs.1.54, P<0.05; caspase-9:2.22vs.1.09, P<0.05).Conclusion:Knock-down of hepcidin induced lymphocyte apoptosis in septic mice, hepcidin may play an inhibitory role on lymphocyte apoptosis in sepsis,...