Hepcidin基因干扰对脓毒症小鼠淋巴细胞凋亡的影响研究

发布时间：2018-05-06 19:22

本文选题：hepcidin + 脓毒血症　；参考：《浙江大学》2014年硕士论文

【摘要】：目的：探讨hepcidin基因干扰对脓毒症小鼠淋巴细胞凋亡的影响,为脓毒症的发病机制和防治提供一定理论参考。方法： BALB/C雄性6-8周健康小鼠10只,随机均分为实验组(hepcidin基因干扰组,n=5)和对照组(RNA干扰腺病毒组,n=5)。实验组小鼠给予尾静脉高动力注射hepcidin特异性siRNA (small interference RNA)腺病毒,对照组小鼠给予尾静脉高动力注射对照的RNA干扰腺病毒,两组小鼠均自由饮食和活动。13天后,两组小鼠采用盲肠结扎穿孔(cecal ligation and puncture, CLP)法复制脓毒症模型。CLP模型制备24小时后处死小鼠,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；TUNEL法检测脾脏组织细胞的凋亡量；Annexin-V染色流式检测外周血和脾脏T淋巴细胞的凋亡百分比；Western blot法检测脾脏组织凋亡相关蛋白caspase-3和caspase-9的活化情况。结果：实验组小鼠脾脏CD4+.T细胞百分比明显低于对照组(17.34%vs.29.08%,P0.05)；实验组小鼠脾脏CD8--T细胞百分比明显低于对照组(8.37%vs.14.57%,P0.05); TUNEL结果显示实验组小鼠脾脏组织细胞凋亡量比对照组小鼠多；实验组小鼠脾脏T淋巴细胞凋亡率较对照组明显增多(79.3%vs.59.6%,P0.05),实验组小鼠外周血T淋巴细胞凋亡率较对照组明显增加(93.0%vs.68.9%,P0.05)；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(11.97vs.1.54, P0.05;2.22vs.1.09, P0.05)。结论： Hepcidin基因干扰引起脓毒症小鼠淋巴细胞凋亡增加,hepcidin可能在脓毒症中对淋巴细胞的凋亡起一定的抑制作用。
[Abstract]:Objective: To investigate the effect of hepcidin gene interference on lymphocyte apoptosis in septic mice, and to provide a theoretical reference for the pathogenesis, prevention and treatment of sepsis. Methods: Ten healthy BALB/C male mice were randomly divided into two groups: the experimental group was treated with hepcidin gene interference and the control group with adenovirus. The mice in the experimental group were injected with hepcidin specific siRNA small interference adenovirus, and the control mice were given RNA interference adenovirus. The mice in both groups were fed free diet and activity for 13 days. The sepsis model was made by cecal ligation and puncture (CLP) method in the two groups. The mice were killed 24 hours later, and the whole blood and spleen samples were collected. Flow cytometry was used to determine the percentage of CD4 and CD8 in spleen T lymphocyte subtypes and Tunel method was used to detect apoptosis of splenic tissue cells. Annexin-V staining flow cytometry was used to detect the percentage of apoptosis in peripheral blood and spleen T lymphocytes. Western blot method was used to detect the percentage of apoptosis in peripheral blood and spleen T lymphocytes. Activation of apoptosis-related proteins caspase-3 and caspase-9 in spleen tissue. Results: The percentage of CD4. T cells in the spleen of the experimental group was significantly lower than that of the control group (17. 34 vs 29. 08), the percentage of the spleen CD8--T cells in the experimental group was significantly lower than that in the control group (8. 37 vs 14. 57). The results of TUNEL showed that the apoptosis of spleen tissue cells in the experimental group was more than that in the control group. Compared with the control group, the apoptosis rate of spleen T lymphocytes in the experimental group was significantly higher than that in the control group. The apoptosis rate of T lymphocytes in the peripheral blood of the experimental group was significantly higher than that of the control group, and the apoptosis-related protein caspase-3 and caspase-9 activated water in the spleen of the experimental group were significantly higher than that of the control group. Compared with the control group, P0.05 + 2.22vs.1.09, P0.05 + 2.22vs.1.09. Conclusion: Apoptosis of lymphocytes in septic mice induced by Hepcidin gene interference may be inhibited by hepcidin in sepsis.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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