辅助性T细胞相关细胞因子在儿童急性免疫性血小板减少性紫癜中的变化及意义
发布时间：2018-10-26 16:19
【摘要】：目的 通过检测急性免疫性血小板减少性紫癜(immune thrombocytopenic purpura ITP)患儿辅助性T(Th)淋巴细胞相关细胞因子γ-干扰素(IFN-γ)、白介素4(IL-4)、B细胞特异单克隆鼠白血病病毒整合位点基因(Bmi-1)及白介素17(IL-17)mRNA表达的变化,分析探讨急性ITP患儿Th细胞相关细胞因子功能变化及其相关性,以期进一步明确Th细胞相关细胞因子在急性ITP患儿发病中的作用。 方法 本实验设为实验组与对照组。实验组36例急性ITP患儿均源于新乡医学院第一附属医院儿科,2012年1月-2012年9月期间住院和门诊患儿,均符合急性ITP诊断标准；对照组为我院同期小儿外科择期手术患儿26例,采集血标本前2周内均无感染史；两组采集血标本前4周内均无输血、糖皮质激素使用及疫苗接种史,均为河南地区汉族儿童,性别、年龄均无统计学差异(P0.05)；均获得其家属知情同意。 采用流式细胞仪(fluorescence-activated cell sorting,FACS)检测实验组与对照组儿童外周血T细胞亚群比例；采用RT-PCR方法检测外周血淋巴细胞中TFN-γ、 Bmi-1、IL-4、IL-17Th相关的细胞因子mRNA的表达；并采用t检验和直线相关分析进行分析比较。 结果 1、实验组CD3+细胞百分比为60.54±3.46,CD4+细胞百分比为30.07±0.79,CD8+细胞百分比为25.23±2.57,CD4+/CD8+细胞比值为1.12±0.19,对照组CD3+细胞百分比为63.73±4.03,CD4+细胞百分比为35.41±4.76,CD8+细胞百分比为21.62±2.53,CD4+/CD8+细胞比值为1.64±0.28；实验组与对照组比较,CD3+细胞低于对照组(t=3.36,P0.001),CD4+细胞明显低于对照组(t=8.30,P0.001),CD8+细胞明显高于对照组(t=5.49,P0.001),CD4+/CD8+明显低于对照组(t=8.72,P0.001)。 2、实验组外周血淋巴细胞中IFN-ymRNA为3.84±0.43, IL-4mRNA为1.44±0.39, Bmi-1mRNA为2.63±0.54,IL-17mRNA为4.69±0.58;对照组外周血淋巴细胞中IFN-ymRNA为2.88±0.57,IL-4mRNA为1.87±0.34,Bmi-1mRNA为3.91±0.92,IL-17mRNA为3.79±0.82；；实验组与对照组比较,IFN-γ明显高于对照组(t=7.56,P0.001),IL-4明显低于对照组(t=4.52,P0.001)；Bmi-1明显低于对照组(t=6.88,P0.001),IL-17明显高于对照组(t=5.07,P0.01)。 3、实验组外周血淋巴细胞中IFN-γ mRNA表达与[L-4mRNA表达呈负相关(r=-0.667,P0.001),实验组外周血淋巴细胞中IL-4mRNA表达与Bmi-1基因mRNA表达呈正相关(r=0.776,P0.001)；实验组外周血淋巴细胞中IFN-γ mRNA表达与IL-17mRNA表达呈正相关(r=0.712,P0.001)；实验组外周血淋巴细胞中IFN-γ mRNA表达与Bmi-1mRNA表达无相关性(r=-0.206, P0.05)；实验组外周血淋巴细胞中IL-17mRNA表达与Bmi-1mRNA表达无相关性(r=-0.214,P0.05)；实验组外周血淋巴细胞中IL-17mRNA表达与IL-4mRNA表达无相关性(r=-0.220,P0.05)。 结论 1急性ITP患儿存在细胞免疫紊乱,Th/Ts细胞失衡,且Th细胞功能紊乱,Th1/Th2比例失调,可能是Th1发挥优势； 2Bmi-1水平降低可能参与急性ITP的发病; 3Th17可能通过促进IL-17的分泌在急性ITP的发病中起作用。
[Abstract]:Objective to detect the cytokines interferon 纬 (IFN- 纬) and interleukin-4 (IL-4) of helper T (Th) lymphocytes in children with acute immune thrombocytopenic purpura (immune thrombocytopenic purpura ITP). The changes of expression of Bmi-1 and IL-17 mRNA in B cell-specific monoclonal mouse leukemia virus (MMLV) were analyzed to investigate the changes of cytokine function of Th cells in children with acute ITP. To further clarify the role of Th cell-related cytokines in the pathogenesis of acute ITP children. Methods the experiment was divided into experimental group and control group. The 36 children with acute ITP in the experimental group were all from paediatrics of the first affiliated Hospital of Xinxiang Medical College. From January 2012 to September 2012, all the hospitalized and outpatient children met the diagnostic criteria of acute ITP. The control group consisted of 26 children undergoing elective operation in pediatric surgery in the same period. There was no history of infection within 2 weeks before blood samples were collected. No blood transfusion, glucocorticoid use and vaccination history were observed in the two groups in the first 4 weeks. There was no significant difference in sex and age between the two groups (P0.05); the informed consent of their families was obtained. The percentage of T lymphocyte subsets in peripheral blood of children in experimental group and control group was detected by flow cytometry (fluorescence-activated cell sorting,FACS). The expression of TFN- 纬 and Bmi-1,IL-4,IL-17Th related cytokines mRNA in peripheral blood lymphocytes was detected by RT-PCR method and compared with t test and linear correlation analysis. Results 1 the percentage of CD3 cells in the experimental group was 60.54 卤3.46, the percentage of CD4 cells was 30.07 卤0.79 and the percentage of CD8 cells was 25.23 卤2.57%. The ratio of CD4 / CD8 cells was 1.12 卤0.19. In the control group, the percentage of CD3 cells was 63.73 卤4.03, the percentage of CD4 cells was 35.41 卤4.76, the percentage of CD8 cells was 21.62 卤2.53, the ratio of CD4 / CD8 cells was 1.64 卤0.28; Compared with the control group, the CD3 cells, CD4 cells and CD8 cells in the experimental group were significantly lower than those in the control group (t = 3.36, P 0.001), and in the control group, the CD8 cells were significantly higher than those in the control group (t = 5.49, P 0.001). The ratio of CD4 / CD8 was significantly lower than that of the control group (t = 8.72, P 0.001). 2. The IFN-ymRNA, IL-4mRNA and Bmi-1mRNA in peripheral blood lymphocytes of the experimental group were 3.84 卤0.43, 1.44 卤0.39 and 2.63 卤0.54 respectively, which were 4.69 卤0.58; In the control group, the level of IFN-ymRNA in peripheral blood lymphocytes was 2.88 卤0.57, IL-4 mRNA was 1.87 卤0.34, Bmi-1 mRNA was 3.91 卤0.92 and IL-17 mRNA was 3.79 卤0.82in peripheral blood lymphocytes. Compared with the control group, the IFN- 纬 of the experimental group was significantly higher than that of the control group (t = 7.56, P 0.001), and the IL-4 was significantly lower than that of the control group (t = 4.52, P 0.001). Bmi-1 was significantly lower than that in the control group (t = 6.88, P 0.001), and IL-17 was significantly higher than that in the control group (t = 5.07, P 0.01). 3. The expression of IFN- 纬 mRNA was negatively correlated with the expression of [L-4mRNA] in peripheral blood lymphocytes of experimental group (r-0.667p0.001), and the expression of IL-4mRNA in peripheral blood lymphocytes of experimental group was positively correlated with the expression of Bmi-1 gene mRNA (r-0.776). (P0.001); There was a positive correlation between the expression of IFN- 纬 mRNA and IL-17mRNA in peripheral blood lymphocytes of the experimental group (r = 0.712, P 0.001), but no correlation between the expression of IFN- 纬 mRNA and the expression of Bmi-1mRNA in the peripheral blood lymphocytes of the experimental group (r = 0.206, P 0.05). There was no correlation between IL-17mRNA expression and Bmi-1mRNA expression in peripheral blood lymphocytes of experimental group (r-0.2145P0.05), but there was no correlation between IL-17mRNA expression and IL-4mRNA expression in peripheral blood lymphocytes of experimental group (r-0.220p0.05). Conclusion (1) Children with acute ITP have cellular immune disorder, Th/Ts cell imbalance, Th cell dysfunction and Th1/Th2 imbalance, which may be the advantage of Th1. The decrease of 2Bmi-1 level may be involved in the pathogenesis of acute ITP and 3Th17 may play a role in the pathogenesis of acute ITP by promoting the secretion of IL-17.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.5
[Abstract]:Objective to detect the cytokines interferon 纬 (IFN- 纬) and interleukin-4 (IL-4) of helper T (Th) lymphocytes in children with acute immune thrombocytopenic purpura (immune thrombocytopenic purpura ITP). The changes of expression of Bmi-1 and IL-17 mRNA in B cell-specific monoclonal mouse leukemia virus (MMLV) were analyzed to investigate the changes of cytokine function of Th cells in children with acute ITP. To further clarify the role of Th cell-related cytokines in the pathogenesis of acute ITP children. Methods the experiment was divided into experimental group and control group. The 36 children with acute ITP in the experimental group were all from paediatrics of the first affiliated Hospital of Xinxiang Medical College. From January 2012 to September 2012, all the hospitalized and outpatient children met the diagnostic criteria of acute ITP. The control group consisted of 26 children undergoing elective operation in pediatric surgery in the same period. There was no history of infection within 2 weeks before blood samples were collected. No blood transfusion, glucocorticoid use and vaccination history were observed in the two groups in the first 4 weeks. There was no significant difference in sex and age between the two groups (P0.05); the informed consent of their families was obtained. The percentage of T lymphocyte subsets in peripheral blood of children in experimental group and control group was detected by flow cytometry (fluorescence-activated cell sorting,FACS). The expression of TFN- 纬 and Bmi-1,IL-4,IL-17Th related cytokines mRNA in peripheral blood lymphocytes was detected by RT-PCR method and compared with t test and linear correlation analysis. Results 1 the percentage of CD3 cells in the experimental group was 60.54 卤3.46, the percentage of CD4 cells was 30.07 卤0.79 and the percentage of CD8 cells was 25.23 卤2.57%. The ratio of CD4 / CD8 cells was 1.12 卤0.19. In the control group, the percentage of CD3 cells was 63.73 卤4.03, the percentage of CD4 cells was 35.41 卤4.76, the percentage of CD8 cells was 21.62 卤2.53, the ratio of CD4 / CD8 cells was 1.64 卤0.28; Compared with the control group, the CD3 cells, CD4 cells and CD8 cells in the experimental group were significantly lower than those in the control group (t = 3.36, P 0.001), and in the control group, the CD8 cells were significantly higher than those in the control group (t = 5.49, P 0.001). The ratio of CD4 / CD8 was significantly lower than that of the control group (t = 8.72, P 0.001). 2. The IFN-ymRNA, IL-4mRNA and Bmi-1mRNA in peripheral blood lymphocytes of the experimental group were 3.84 卤0.43, 1.44 卤0.39 and 2.63 卤0.54 respectively, which were 4.69 卤0.58; In the control group, the level of IFN-ymRNA in peripheral blood lymphocytes was 2.88 卤0.57, IL-4 mRNA was 1.87 卤0.34, Bmi-1 mRNA was 3.91 卤0.92 and IL-17 mRNA was 3.79 卤0.82in peripheral blood lymphocytes. Compared with the control group, the IFN- 纬 of the experimental group was significantly higher than that of the control group (t = 7.56, P 0.001), and the IL-4 was significantly lower than that of the control group (t = 4.52, P 0.001). Bmi-1 was significantly lower than that in the control group (t = 6.88, P 0.001), and IL-17 was significantly higher than that in the control group (t = 5.07, P 0.01). 3. The expression of IFN- 纬 mRNA was negatively correlated with the expression of [L-4mRNA] in peripheral blood lymphocytes of experimental group (r-0.667p0.001), and the expression of IL-4mRNA in peripheral blood lymphocytes of experimental group was positively correlated with the expression of Bmi-1 gene mRNA (r-0.776). (P0.001); There was a positive correlation between the expression of IFN- 纬 mRNA and IL-17mRNA in peripheral blood lymphocytes of the experimental group (r = 0.712, P 0.001), but no correlation between the expression of IFN- 纬 mRNA and the expression of Bmi-1mRNA in the peripheral blood lymphocytes of the experimental group (r = 0.206, P 0.05). There was no correlation between IL-17mRNA expression and Bmi-1mRNA expression in peripheral blood lymphocytes of experimental group (r-0.2145P0.05), but there was no correlation between IL-17mRNA expression and IL-4mRNA expression in peripheral blood lymphocytes of experimental group (r-0.220p0.05). Conclusion (1) Children with acute ITP have cellular immune disorder, Th/Ts cell imbalance, Th cell dysfunction and Th1/Th2 imbalance, which may be the advantage of Th1. The decrease of 2Bmi-1 level may be involved in the pathogenesis of acute ITP and 3Th17 may play a role in the pathogenesis of acute ITP by promoting the secretion of IL-17.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.5
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