基于B、T淋巴细胞衰减因子及氧化应激探讨新风胶囊治疗强直性脊柱炎的作用机制

Exploration of the Mechanism of Xinfeng Capsule in the Treatment of Ankylosing Spondylitis Based on B and T Lymphocyte Attenuator and Oxidative Stress

目的探讨强直性脊柱炎(AS)患者B、T淋巴细胞衰减因子(BTLA)及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、总抗氧化能力(TAOC)、活性氧(ROS)、活性氮(RNS)、丙二醛(MDA)的变化及新风胶囊(XFC)对其影响。方法将120例AS患者按随机数字表法分为两组:XFC组(60例)和柳氮磺胺吡啶(SASP)组(60例)。两组均治疗3个月。60名健康体检者为健康对照组(NC组)。采用流式细胞术检测患者外周血BTLA表达频率及活化水平;采用酶联免疫分析法检测两组血清中氧化应激指标(SOD、CAT、TAOC、ROS、RNS、MDA)和肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、IL-4、IL-10含量;采用魏氏法检测炎性指标血沉(ESR);采用日立7060型全自动生化分析仪检测超敏C反应蛋白(Hs-CRP);采用VAS及ASAS 20、BASDAI 50评定疗效;并进行生活质量评分及BTLA表达频率的相关分析。结果(1)与SASP组比较,XFC组ASAS 20及BASDAI 50疗效更优,差异有统计学意义(P<0.01)。(2)与NC组比较,AS患者外周血BTLA表达显著降低(P<0.05);SOD、CAT、TAOC值显著降低(P<0.01,P<0.05);ROS、RNS、MDA值显著升高(P<0.01,P<0.05);TNF-α、IL-1β、ESR、Hs-CRP值显著升高(P<0.01),IL-4、IL-10值显著降低(P<0.01,P<0.05)。(3)与本组治疗前比较,治疗后两组外周血BTLA/CD19+B、BTLA/CD24+B、SOD、TAOC、IL-4、SF-36量表8个维度积分值均显著升高,ROS、MDA、TNF-α、ESR、Hs-CRP、VAS、BASDAI 50、BASFI、BAS-G均显著降低(P<0.01,P<0.05)。XFC组在升高BTLA/CD19+B、BTLA/CD24+B、SOD、TAOC、IL-10、BP、MH、VT、SF及降低ROS、IL-1β、MDA、TNF-α、ESR、Hs-CRP、VAS、BASDAI、BASFI、BAS-G方面优于SASP组(P<0.01,P<0.05)。(4)Pearson相关分析结果显示:外周血BTLA/CD19+B与SOD、CAT、TAOC、IL-4、IL-10、GH、RP、BP、SF呈正相关(r分别为0.431,0.325,0.318,0.316,0.348,0.314,0.358,0.318,0.326;P<0.05,P<0.01),与ROS、MDA、TNF-α、IL-1β、ESR、VAS、BASDAI呈负相关(r分别为-0.342;-0.368,-0.334,-0.354,-0.324,-0.372,-0.342,P<0.05,P<0.01)。BTLA/CD24+B与SOD、TAOC、IL-4、IL-10、GH、RP、BP、SF、RE、MH、VT亦呈正相关(r分别为0.358,0.352,0.372,0.436,0.435,0.326,0.352,0.345,0.326,0.343,0.332;P<0.05,P<0.01),与ROS、RNS、MDA、ESR、Hs-CRP、VAS、BASDAI、BASFI呈负相关(r分别为-0.447,-0.336,-0.405,-0.395,-0.358,-0.436,-0.338,-0.425;P<0.05,P<0.01)。结论 XFC能提高AS患者外周血BTLA表达,负性调节B细胞的激活与增殖,降低异常免疫反应和氧化应激损伤,从而有效减轻关节僵痛症状。

Objective To explore changes of B and T lymphocyte attenuator（ BTLA）,superoxide dismutase（ SOD）,catalase（ CAT）,total antioxidant capacity（ TAOC）,reactive oxygen species（ ROS）,reactive nitrogen species（ RNS）,malondialdehyde（ MDA） in ankylosing spondylitis（ AS） patients,and the effect of Xinfeng Capsule（ XFC） on them. Methods Totally 120 AS patients were assigned to two groups according to random digit table method,the XFC group（ 3 XFC pills each time,3 times a day） and the SASP group（ 4 SASP tablets each time,twice a day）,60 in each group. All patients were treated for 3 months. Another 60 healthy subjects were recruited as a healthy control group. The expression frequency and activation levels of BTLA were detected using flow cytometry. Serum oxidative stress indices（ such as SOD and CAT,TAOC,ROS,RNS,MDA） and contents of cytokines [tumor necrosis factor α（ TNF-α）,IL-1 β,IL-4,and IL-10] were detected using enzyme-linked immunoassay（ ELISA）. Erythrocyte sedimentation rate（ ESR） was detected using Westergren method. High-sensitivity C-reactive protein（ HsCRP） was detected using HITACHI 7060 type automatic biochemical analyzer. Clinical efficacies of ASAS 20 and BASDAI50 were assessed using VAS. Correlation analysis between scoring for quality of life and BTLA expression frequency was performed. Results（ 1） Clinical efficacies of ASAS 20 and BASDAI50 were significantly better in the XFC group than in the SASP group（P 〈 0. 01）.（ 2） Compared with the healthy control group,BTLA expressions in the peripheral blood of AS patients decreased significantly（ P 〈 0. 05）; SOD,CAT,and TAOC values significantly decreased（P 〈 0. 01,P 〈 0. 05）; ROS,RNS,and MDA values significantly increased（ P 〈 0. 01,P 〈 0. 05）;TNF-α,IL-1 β,ESR,and Hs-CRP values significantly increased（P 〈 0. 01）; IL-4 and IL-10 values decreased significantly（P 〈 0. 01,P 〈 0. 05）.（ 3） Compared with pre-treatment in the same group,BTLA / CD19＋B,BTLA /CD24＋B,SOD,TAOC,IL-4,SF-36 [physical functioning（ PF）,social functioning（ SF）,role limitation due to physical problems（ RP）,role limitation due to emotional problems（ RE）,body pain（ BP）,mental health（ MH）,vitality（ VT）,general health（ GH） ] were significantly elevated; ROS,MDA,TNF-α,ESR,Hs-CRP,VAS,BASDAI and BASFI,BAS-G were significantly lower in the peripheral blood of the two groups after treatment（P 〈 0. 01,P〈 0. 05）. Better effect was shown in the XFC group in elevating BTLA / CD19＋B,BTLA / CD24＋B,SOD,TAOC,IL-10,BP,MH,VT,and SF; and lowering ROS,IL-1 β,MDA,TNF-α,ESR,Hs-CRP,VAS,BASDAI,BASFI,and BAS-G（P 〈 0. 01,P 〈 0. 05）.（ 4） Pearson correlation analysis showed,BTLA / CD19＋B expression of the peripheral blood was positively correlated with SOD,CAT,TAOC,IL-4,IL-10,GH,RP,BP,and SF（ r = 0. 431,0. 325,0. 318,0. 316,0. 348,0. 314,0. 358,0. 318,0. 326,respectively,P 〈 0. 05,P 〈 0. 01）,while it was negative correlated with ROS,MDA,TNF-α,IL-1 β,ESR,VAS,and BASDAI（ r =- 0. 342,- 0. 368,-0. 334,- 0. 354,- 0. 324,- 0. 372,- 0. 342,respectively,P 〈 0. 05,P 〈 0. 01）. BTLA / CD24＋B expression of the peripheral blood was positively correlated with SOD,TAOC,IL-4,IL-10,GH,RP,BP,SF,RE,MH,VT（r= 0. 358,0. 352,0. 372,0. 436,0. 435,0. 326,0. 352,0. 345,0. 326,0. 343,0. 332,respectively,P〈 0. 05,P 〈 0. 01）,while it was negative correlated with ROS,RNS,MDA,ESR,Hs-CRP,VAS,BASDAI,and BASFI（r =- 0. 447,- 0. 336,- 0. 405,- 0. 395,- 0. 358,- 0. 436,- 0. 338,- 0. 425,respectively,P 〈0. 05,P 〈 0. 01）. Conclusion XFC could improve BTLA expression in the peripheral blood of AS patients,negatively regulate activation and proliferation of B cells,and reduce abnormal immune responses and oxidative stress injury,thereby effectively alleviating joint stiffness and pain.