CAJ | 학술논문

目的：探讨 CD4+ T 淋巴细胞亚群及相应转录因子和上游转录调节因子在过敏性紫癜（HSP）病理机制中的调控作用。方法采用流式细胞术（FCM）、酶联免疫吸附法（ ELISA）、荧光定量聚合酶链反应（RT-PCR）测定41例 HSP 急性期患儿（急性期组）、35例 HSP 临床缓解期患儿（缓解组）和30例健康儿童（健康对照组）外周血单个核细胞（PBMC）中辅助性 T 淋巴细胞（Th）1、Th2、调节性 T 淋巴细胞（Treg）、Th17细胞百分比，血浆干扰素-γ（IFN-γ）、白细胞介素（IL）-4、转化生长因子-β1（TGF-β1）、IL-17水平和 PBMC 中 T 淋巴细胞 T-box（ T-bet）、GATA 结合蛋白-3（ GATA3）、插状头/翅膀状螺旋转录因子-3（ Foxp3）和孤核受体γ（ROR-γt）mRNA 表达。结果1. HSP 急性期组、缓解组患儿 PBMC 中 Th1、Treg 细胞百分比，相应血浆 IFN-γ、TGF-β1水平，T-bet、Foxp3 mRNA 表达均低于健康对照组（P 均﹤0.05），Th2、Th17细胞百分比，血浆 IL-4、IL-17水平和 GATA3、ROR-γt mRNA 表达均高于健康对照组（P 均﹤0.05），Th1/ Th2和 Treg/ Th17失衡；2. HSP 缓解组患儿 PBMC 中 Th1、Th2、Th17、Treg 和 Treg/ Th17与急性期组比较差异均无统计学意义（ P 均﹥0.05）， Th1/ Th2、TGF-β1及相应 T-bet mRNA、Foxp3 mRNA 高于急性期组（P 均﹤0.05），IL-4、IL-17及相应 GATA3、ROR-γt mRNA 均低于急性期组（P 均﹤0.05），Th1/ Th2和 Treg/ Th17仍存在失衡；3. Th1、Th2、Treg、Th17百分数分别与 IFN-γ、IL-4、TGF-β1、IL-17，T-bet、GATA3、Foxp3和 ROR-γt mRNA 表达均呈正相关（ P 均﹤0.05）。结论 HSP 存在 Th1/ Th2、Treg/ Th17免疫失衡，Treg 诱导的免疫耐受被打破，并贯穿整个病理过程。
목적：탐토 CD4+ T 림파세포아군급상응전록인자화상유전록조절인자재과민성자전（HSP）병리궤제중적조공작용。방법채용류식세포술（FCM）、매련면역흡부법（ ELISA）、형광정량취합매련반응（RT-PCR）측정41례 HSP 급성기환인（급성기조）、35례 HSP 림상완해기환인（완해조）화30례건강인동（건강대조조）외주혈단개핵세포（PBMC）중보조성 T 림파세포（Th）1、Th2、조절성 T 림파세포（Treg）、Th17세포백분비，혈장간우소-γ（IFN-γ）、백세포개소（IL）-4、전화생장인자-β1（TGF-β1）、IL-17수평화 PBMC 중 T 림파세포 T-box（ T-bet）、GATA 결합단백-3（ GATA3）、삽상두/시방상라선전록인자-3（ Foxp3）화고핵수체γ（ROR-γt）mRNA 표체。결과1. HSP 급성기조、완해조환인 PBMC 중 Th1、Treg 세포백분비，상응혈장 IFN-γ、TGF-β1수평，T-bet、Foxp3 mRNA 표체균저우건강대조조（P 균﹤0.05），Th2、Th17세포백분비，혈장 IL-4、IL-17수평화 GATA3、ROR-γt mRNA 표체균고우건강대조조（P 균﹤0.05），Th1/ Th2화 Treg/ Th17실형；2. HSP 완해조환인 PBMC 중 Th1、Th2、Th17、Treg 화 Treg/ Th17여급성기조비교차이균무통계학의의（ P 균﹥0.05）， Th1/ Th2、TGF-β1급상응 T-bet mRNA、Foxp3 mRNA 고우급성기조（P 균﹤0.05），IL-4、IL-17급상응 GATA3、ROR-γt mRNA 균저우급성기조（P 균﹤0.05），Th1/ Th2화 Treg/ Th17잉존재실형；3. Th1、Th2、Treg、Th17백분수분별여 IFN-γ、IL-4、TGF-β1、IL-17，T-bet、GATA3、Foxp3화 ROR-γt mRNA 표체균정정상관（ P 균﹤0.05）。결론 HSP 존재 Th1/ Th2、Treg/ Th17면역실형，Treg 유도적면역내수피타파，병관천정개병리과정。
Objective To explore the alternations of CD4 + T cell subsets and their cytokines and transcription factors in children with Henoch - Schonlein purpura(HSP). Methods Forty - one children were enrolled in this stud-y,including 41 in acute stage of HSP,35 cases of HSP in clinical remission stage and 30 healthy children as healthy control group. The percentages of helper T lymphocytes(Th)1,Th2,regulatory T cells(Treg)and Th17 subsets were determined by flow cytometry(FCM). The concentrations of plasma interferon - γ( INF - γ),interleukin( IL) - 4, transforming growth factor - β1(TGF - β1 )and IL - 17 were examined by ways of enzyme - linked immunosorbent assay(ELISA). The expressions of T - bet,GATA3,Foxp3 and ROR - γt mRNA in peripheral blood mononuclear cells were examined by means of reverse transcription - polymerase chain reaction(RT - PCR). Results (1)During the a-cute and recovery stage of HSP,compared with the control group,the percentages of Th2 and Th17 cell subsets,the le-vels of plasma IL - 4 and IL - 17,and the expressions of GATA3 and ROR - γt mRNA were significantly higher(all P ﹤ 0. 05). The percentages of Th1 and Treg cell subsets,the levels of plasma INF - γ and TGF - β1 and the expres-sions of T - bet and Foxp3 mRNA were lower(all P ﹤ 0. 05). The imbalances of Th1 / Th2 and Treg/ Th17 appeared during the acute stage of HSP.(2)During the recovery stage of HSP,the percentages of Th1,Th2,Th17 and Treg,and the ratio of Treg / Th17 were of no difference compared with those in acute stage( all P ﹥ 0. 05). The ratio of Th1 / Th2,and the expressions of T - bet and Foxp3 mRNA were increased(all P ﹤ 0. 05),while the levels of plasma IL - 4 and IL - 17,the expressions of GATA3 and ROR - γt mRNA were decreased(all P ﹤ 0. 05)compared with those in acute stage. In the recovery stage of HSP,the imbalances of Th1 / Th2 and Treg/ Th17 were still obvious .(3)There was a positive correlation in every 2 cytokines of Th1,INF - γ and T - bet. And the same correlation existed in Th2, IL - 4 and GATA3,in Treg,TGF - β1 and Foxp3,and in Th17,IL - 17 and ROR - γt(P ﹤ 0. 05). Conclusions The imbalance of Th1 / Th2 and Treg/ Th17 is critical in pathological mechanism of HSP. The disturbance of immune tole-rance induced by Treg cells is important in HSP.