强直性脊柱炎患者单个核细胞TLR2、TLR4与淋巴细胞凋亡的研究

Study of TLR2, TLR4 and Apoptosis of Lymphocytes in Patients with Ankylosing Spondylitis

摘要: 目的：通过检测强直性脊柱炎(ankylosing spondylitis, AS)患者外周血单个核细胞(单核细胞、淋巴细胞)表面TLR2 (Toll-like receptors2, TLR2)、TLR4 (Toll-like receptors4, TLR4)的表达及淋巴细胞凋亡的数量，探讨TLR2、TLR4及淋巴细胞的凋亡与强直性脊柱炎发生发展的关系。方法：应用流式细胞术检测67例AS患者(非活动性组39例，活动性组28例)和30例健康体检者的外周血标本。通过应用FITC-TLR2、PE-TLR4两种试剂检测外周血单个核细胞表面TLR2、TLR4的表达水平,以及流式Annexin-v FITC/PI法检测外周血淋巴细胞的凋亡量。各实验检测均设定相应的同型或阴性对照。结果：1) AS患者外周血单个核细胞表面TLR2在疾病组、活动性组和非活动性组分别为：23.16%、23.07%和23.22%，均高于健康对照组12.53%，差异有统计学意义(P < 0.01)；AS非活动性组与活动性组比较差异无统计学意义(P > 0.05)。2) AS患者外周血单个核细胞表面TLR4在疾病组、活动性组和非活动性组分别为：24.43%、24.51%和24.38%，均高于健康对照组12.55%，差异有统计学意义(P < 0.01)；AS非活动性组与活动性组比较差异无统计学意义(P > 0.05)。3) AS患者外周血淋巴细胞凋亡在疾病组、活动性组、非活动性组分别为： 84.00%、79.64%和87.14%，均低于健康对照组98.26%，差异有统计学意义(P < 0.01)；AS活动性组低于非活动性组，差异有统计学意义(P < 0.01)。4) AS患者外周血单个核细胞表面TLR2、TLR4与AS活动性(BASDI)无相关性(r = 0.043及0.112，P > 0.05)；AS患者外周血淋巴细胞凋亡与活动性呈中度负相关(r = −0.568, P < 0.05)。结论：1) AS患者外周血单个核细胞表面TLR2、TLR4出现异常增高。2) AS患者外周血淋巴细胞凋亡量减少。

Abstract: Objective: By detecting the expression of TLR2 (TLR2) and TLR4 (TLR4) on the surface of peripheral blood mononuclear cells (mononuclear cells and lymphocytes) and the number of lymphocyte apoptosis in patients with ankylosing spondylitis (AS), the relationship between the apoptosis of TLR2, TLR4 and lymphocyte and the occurrence and development of ankylosing spondylitis was discussed. Methods: Peripheral blood samples of 67 AS patients (39 in the inactive group and 28 in the active group) and 30 healthy subjects were detected by flow cytometry. Expression levels of TLR2 and TLR4 on the surface of peripheral blood mononuclear cells were detected by fitc-tlr2 and pe-tlr4 reagents, and apoptosis of peripheral blood lymphocytes was measured by flow annexin-v FITC/PI method. Corresponding homotypic or negative controls were set for each test. Results: 1) TLR2 on the surface of peripheral blood mononuclear cells in patients with AS was 23.16% in the disease group, 23.07% in the active group and 23.22% in the inactive group, respectively, higher than 12.53% in the healthy control group, and the difference was statistically significant (P < 0.01). There was no statistically significant difference between the inactive AS group and the active AS group (P > 0.05). 2) TLR4 on the surface of peripheral blood mononuclear cells in patients with AS was 24.43%, 24.51% and 24.38% respectively in the disease group, the active group and the inactive group, all higher than 12.55% in the healthy control group, and the difference was statistically significant (P < 0.01). There was no statistically significant difference between the inactive AS group and the active AS group (P > 0.05). 3) The apoptosis of peripheral blood lymphocytes in patients with AS was 84.00% in the disease group, 79.64% in the active group and 87.14% in the inactive group, respectively, which was lower than that in the healthy control group (98.26%), and the difference was statistically significant (P < 0.01). AS activity group was lower than the inactive group, and the difference was statistically significant (P < 0.01). 4) TLR2 and TLR4 on the surface of mononuclear cells in peripheral blood of AS patients were not correlated with AS activity (BASDI) (r = 0.043 and 0.112, P > 0.05). Peripheral blood lymphocyte apoptosis was negatively correlated with activity in patients with AS (r = −0.568, P < 0.05). Conclusion: 1) Abnormal increase of TLR2 and TLR4 were appeared on the surface of mononuclear cells in peripheral blood of AS patients. 2) Apoptosis of lymphocytes in peripheral blood of AS patients decreased.