CAJ | 학술논문

目的探讨强直性脊柱炎(AS)患者外周血CD4+T细胞白细胞介素(IL)-17蛋白水平和IL-17、核孤儿受体(RORγt)mRNA水平及意义.方法取AS患者和健康人外周血单个核细胞(PBMCs),免疫磁珠分选CD4+T细胞,加佛波脂(PMA)腐子霉素(Ion)作用,经固定/透膜处理进行细胞内染色,流式细胞术(FCM)检测CD4+T细胞IL-17蛋白水平;用或不用非特异性刺激剂[抗CD3单抗(A-CD3)、抗CD28单抗(A-CD28)]刺激CIM+T细胞12 h和24 h后,应用实时荧光定量反转录聚合酶链反应(RT-PCR)检测IL-17、RORγt mRNA水平.组间比较采用方差分析,相关性分析采用Pearson相关分析.结果 CD4+T细胞IL-17蛋白水平AS活动组(1.26±0.44)较稳定组(0.56±0.21)和对照组(0.45±0.13)显著增高(P<0.01),同时IL-17、RORγt mRNA水平活动组较稳定组和对照组显著增高(P<0.01). 用A-CD3/A-CD28刺激后,AS患者CD4+T细胞IL-17、RORγtmRNA水平显著高于未刺激组(P<0.05),12 h刺激组又高于24 h刺激组(P<0.01).结论 AS患者外周血CD4+T细胞IL-17、RORγt表达水平显著增高,IL-17、RORγt表达水平可能与AS病情活动相关.
목적 탐토강직성척주염(AS)환자외주혈CD4+T세포백세포개소(IL)-17단백수평화IL-17、핵고인수체(RORγt)mRNA수평급의의.방법 취AS환자화건강인외주혈단개핵세포(PBMCs),면역자주분선CD4+T세포,가불파지(PMA)부자매소(Ion)작용,경고정/투막처리진행세포내염색,류식세포술(FCM)검측CD4+T세포IL-17단백수평;용혹불용비특이성자격제[항CD3단항(A-CD3)、항CD28단항(A-CD28)]자격CIM+T세포12 h화24 h후,응용실시형광정량반전록취합매련반응(RT-PCR)검측IL-17、RORγt mRNA수평.조간비교채용방차분석,상관성분석채용Pearson상관분석.결과 CD4+T세포IL-17단백수평AS활동조(1.26±0.44)교은정조(0.56±0.21)화대조조(0.45±0.13)현저증고(P<0.01),동시IL-17、RORγt mRNA수평활동조교은정조화대조조현저증고(P<0.01). 용A-CD3/A-CD28자격후,AS환자CD4+T세포IL-17、RORγtmRNA수평현저고우미자격조(P<0.05),12 h자격조우고우24 h자격조(P<0.01).결론 AS환자외주혈CD4+T세포IL-17、RORγt표체수평현저증고,IL-17、RORγt표체수평가능여AS병정활동상관.
Objective To study the expression of IL-17, RORγt in CD4+T cells from patients with ankylosing spondylitis (AS). Methods The specimens of venous blood PBMC were collected from 28 patients with AS and 15 healthy subjects. Intracellular flow cytometry detection of IL-17 was established after isolation of human CD4+ T cells from PBMC. The expression level of IL-17, RORγt mRNA in CD4+ T cells was determined from 28 AS patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti -CD3/Anti -CD28 as stimulators or not. Analysis of variance and Pearson correlation were selected. Results The isolation of human CD4+ T cells from PBMC was effective and its purity reached 90%. The percentage of intracellular IL-17 in CD4+ T cells from AS pati-ents in the AS active group was higher than that of the AS stable group and healthy control group (P<0.01). The expression level of IL-17, RORγt mRNA in CD4+ T cells was significantly higher in patients with AS than in controls. After stimulated with anti-CD3/ anti-CD28 stimulation, the percentage of IL-17, RORγt mRNA was increased significantly (P<0.01). The percentage of IL-17, RORγt mRNA in the 12 h group was higher than that of the 24 h group, while both of them were higher than those without stimulation (P<0.05). Conclusion There is an abnormal expression of IL-17, RORγt in human CD4+ T cells from AS patients. Our results indicate that the abnormal expression of IL-17 might play a role in the development and progression of AS.