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目的观察强直性脊柱炎(AS)患者外周血白介素(IL)-17A、IL-23水平及Th17/Treg比例,探讨骨髓间充质干细胞(BMSCs)与Th17/Treg体外相互作用,为研究AS发生机制提供理论依据。方法研究对象为48例活动期AS患者[实验组,男43例、女5例,年龄(26.2±5.2)岁,均为HLA-B27~+]和49例健康志愿者[对照组,男44例、女5例,年龄(25.9±4.8)岁,HLA-B27~+43例、HLA-B27~-6例]。ELISA检测外周血清中IL-17A及IL-23水平,流式细胞术检测外周血单个核细胞(PBMCs)中的CCR4~+CCR6~+Th细胞(Th17)及Treg细胞数量。分离健康志愿者的PBMCs,将PBMCs分别与AS患者及健康志愿者的BMSCs体外共培养72 h,收集PBMCs行流式细胞术检测,评价BMSCs与Th17/Treg体外的相互作用。结果实验组与对照组血清IL-17A及IL-23的表达水平相近(P>0.05);实验组Th17细胞明显高于对照组(P<0.05),而Treg细胞明显低于对照组(P<0.05)。与健康志愿者BMSCs共培养72 h后的PBMCs中Th17较培养前轻度下降,Treg轻度上升,差异无统计学意义(P>0.05);而与AS患者BMSCs共培养72 h的PBMCs中Th17较培养前及对照组均明显上升,Treg均明显下降,差异均有统计学意义(P<0.05)。结论 AS患者PBMCs中Th17/Treg细胞亚群失衡。免疫抑制能力明显下降的BMSCs极可能通过诱导Th17/Treg失衡而在AS免疫发生机制中发挥重要作用。
Objective To observe the levels of interleukin (IL) -17A and IL-23 in peripheral blood and the ratio of Th17 / Treg in patients with ankylosing spondylitis (AS), and to explore the in vitro interaction between bone marrow mesenchymal stem cells (BMSCs) and Th17 / Mechanism to provide a theoretical basis. METHODS: Forty-eight patients with active AS (experimental group, 43 males and 5 females, aged 26.2 ± 5.2 years, all HLA-B27 +) and 49 healthy volunteers (control group, male 44 5 cases of female, age (25.9 ± 4.8) years, HLA-B27 ~ +43 cases, HLA-B27 ~ -6 cases]. The levels of IL-17A and IL-23 in peripheral blood were detected by ELISA. The numbers of CCR4 + CCR6 + Th cells (Th17) and Treg cells in peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry. PBMCs were isolated from healthy volunteers. PBMCs were co-cultured with AS and healthy volunteers respectively for 72 h in vitro. PBMCs were collected and examined by flow cytometry to evaluate the interaction between BMSCs and Th17 / Treg in vitro. Results The levels of IL-17A and IL-23 in serum of experimental group and control group were similar (P> 0.05). Th17 cells in experimental group were significantly higher than those in control group (P <0.05), while Treg cells were significantly lower in control group (P < 0.05). Compared with the healthy volunteers BMSCs co-cultured for 72 h, the levels of Th17 in PBMCs slightly decreased and Tregs slightly increased (P> 0.05). However, the levels of Th17 in PBMCs co-cultured with BMSCs of AS patients for 72 h Compared with the pre-culture group and the control group, the Tregs were significantly increased, the differences were statistically significant (P <0.05). Conclusion The imbalance of Th17 / Treg subsets in peripheral blood mononuclear cells from AS patients. BMSCs with significantly decreased immunosuppressive capacity play an important role in the pathogenesis of AS probably by inducing imbalance of Th17 / Treg.