【摘要】：目的 通過檢測(cè)急性免疫性血小板減少性紫癜(immune thrombocytopenic purpura ITP)患兒輔助性T(Th)淋巴細(xì)胞相關(guān)細(xì)胞因子γ-干擾素(IFN-γ)、白介素4(IL-4)、B細(xì)胞特異單克隆鼠白血病病毒整合位點(diǎn)基因(Bmi-1)及白介素17(IL-17)mRNA表達(dá)的變化,分析探討急性ITP患兒Th細(xì)胞相關(guān)細(xì)胞因子功能變化及其相關(guān)性,以期進(jìn)一步明確Th細(xì)胞相關(guān)細(xì)胞因子在急性ITP患兒發(fā)病中的作用。 方法 本實(shí)驗(yàn)設(shè)為實(shí)驗(yàn)組與對(duì)照組。實(shí)驗(yàn)組36例急性ITP患兒均源于新鄉(xiāng)醫(yī)學(xué)院第一附屬醫(yī)院兒科,2012年1月-2012年9月期間住院和門診患兒,均符合急性ITP診斷標(biāo)準(zhǔn)；對(duì)照組為我院同期小兒外科擇期手術(shù)患兒26例,采集血標(biāo)本前2周內(nèi)均無(wú)感染史；兩組采集血標(biāo)本前4周內(nèi)均無(wú)輸血、糖皮質(zhì)激素使用及疫苗接種史,均為河南地區(qū)漢族兒童,性別、年齡均無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)；均獲得其家屬知情同意。 采用流式細(xì)胞儀(fluorescence-activated cell sorting,FACS)檢測(cè)實(shí)驗(yàn)組與對(duì)照組兒童外周血T細(xì)胞亞群比例；采用RT-PCR方法檢測(cè)外周血淋巴細(xì)胞中TFN-γ、 Bmi-1、IL-4、IL-17Th相關(guān)的細(xì)胞因子mRNA的表達(dá)；并采用t檢驗(yàn)和直線相關(guān)分析進(jìn)行分析比較。 結(jié)果 1、實(shí)驗(yàn)組CD3+細(xì)胞百分比為60.54±3.46,CD4+細(xì)胞百分比為30.07±0.79,CD8+細(xì)胞百分比為25.23±2.57,CD4+/CD8+細(xì)胞比值為1.12±0.19,對(duì)照組CD3+細(xì)胞百分比為63.73±4.03,CD4+細(xì)胞百分比為35.41±4.76,CD8+細(xì)胞百分比為21.62±2.53,CD4+/CD8+細(xì)胞比值為1.64±0.28；實(shí)驗(yàn)組與對(duì)照組比較,CD3+細(xì)胞低于對(duì)照組(t=3.36,P0.001),CD4+細(xì)胞明顯低于對(duì)照組(t=8.30,P0.001),CD8+細(xì)胞明顯高于對(duì)照組(t=5.49,P0.001),CD4+/CD8+明顯低于對(duì)照組(t=8.72,P0.001)。 2、實(shí)驗(yàn)組外周血淋巴細(xì)胞中IFN-ymRNA為3.84±0.43, IL-4mRNA為1.44±0.39, Bmi-1mRNA為2.63±0.54,IL-17mRNA為4.69±0.58;對(duì)照組外周血淋巴細(xì)胞中IFN-ymRNA為2.88±0.57,IL-4mRNA為1.87±0.34,Bmi-1mRNA為3.91±0.92,IL-17mRNA為3.79±0.82；；實(shí)驗(yàn)組與對(duì)照組比較,IFN-γ明顯高于對(duì)照組(t=7.56,P0.001),IL-4明顯低于對(duì)照組(t=4.52,P0.001)；Bmi-1明顯低于對(duì)照組(t=6.88,P0.001),IL-17明顯高于對(duì)照組(t=5.07,P0.01)。 3、實(shí)驗(yàn)組外周血淋巴細(xì)胞中IFN-γ mRNA表達(dá)與[L-4mRNA表達(dá)呈負(fù)相關(guān)(r=-0.667,P0.001),實(shí)驗(yàn)組外周血淋巴細(xì)胞中IL-4mRNA表達(dá)與Bmi-1基因mRNA表達(dá)呈正相關(guān)(r=0.776,P0.001)；實(shí)驗(yàn)組外周血淋巴細(xì)胞中IFN-γ mRNA表達(dá)與IL-17mRNA表達(dá)呈正相關(guān)(r=0.712,P0.001)；實(shí)驗(yàn)組外周血淋巴細(xì)胞中IFN-γ mRNA表達(dá)與Bmi-1mRNA表達(dá)無(wú)相關(guān)性(r=-0.206, P0.05)；實(shí)驗(yàn)組外周血淋巴細(xì)胞中IL-17mRNA表達(dá)與Bmi-1mRNA表達(dá)無(wú)相關(guān)性(r=-0.214,P0.05)；實(shí)驗(yàn)組外周血淋巴細(xì)胞中IL-17mRNA表達(dá)與IL-4mRNA表達(dá)無(wú)相關(guān)性(r=-0.220,P0.05)。 結(jié)論 1急性ITP患兒存在細(xì)胞免疫紊亂,Th/Ts細(xì)胞失衡,且Th細(xì)胞功能紊亂,Th1/Th2比例失調(diào),可能是Th1發(fā)揮優(yōu)勢(shì)； 2Bmi-1水平降低可能參與急性ITP的發(fā)病; 3Th17可能通過促進(jìn)IL-17的分泌在急性ITP的發(fā)病中起作用。
[Abstract]:Objective to detect the cytokines interferon 緯 (IFN- 緯) 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- 緯 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 鹵3.46, the percentage of CD4 cells was 30.07 鹵0.79 and the percentage of CD8 cells was 25.23 鹵2.57%. The ratio of CD4 / CD8 cells was 1.12 鹵0.19. In the control group, the percentage of CD3 cells was 63.73 鹵4.03, the percentage of CD4 cells was 35.41 鹵4.76, the percentage of CD8 cells was 21.62 鹵2.53, the ratio of CD4 / CD8 cells was 1.64 鹵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 鹵0.43, 1.44 鹵0.39 and 2.63 鹵0.54 respectively, which were 4.69 鹵0.58; In the control group, the level of IFN-ymRNA in peripheral blood lymphocytes was 2.88 鹵0.57, IL-4 mRNA was 1.87 鹵0.34, Bmi-1 mRNA was 3.91 鹵0.92 and IL-17 mRNA was 3.79 鹵0.82in peripheral blood lymphocytes. Compared with the control group, the IFN- 緯 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- 緯 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- 緯 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- 緯 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.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R725.5

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