基于Th17/Treg/Th1免疫失衡探讨长蛇灸干预强直性脊柱炎的疗效及机制研究
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摘要
目的:探讨长蛇灸干预强直性脊柱炎(AS)的疗效及作用机制。方法:60例AS患者随机分为观察组和对照组,每组30例。观察组给予长蛇灸疗法,一组穴以督脉穴大椎至腰俞为主,另一组穴以两侧夹脊穴为主,两组交替使用,每日灸1次,连续治疗7 d为一疗程,疗程间间隔3 d,共治疗4个疗程;对照组给予西药常规治疗,即柳氮磺胺吡啶配合非甾体抗炎药,连续服药7 d为一疗程,治疗4个疗程。采用酶联免疫吸附实验(ELISA)法检测血浆白细胞介素-6(IL-6)、白细胞介素-17(IL-17)、白细胞介素-23(IL-23)及肿瘤坏死因子α(TNF-α)水平;采用实时定量聚合酶链反应(RT-PCR)检测辅助性17细胞(Th17)、调节性T细胞(Treg)、T辅助细胞1型(Th1)的特异性转录因子Fox P3、T-bet的m RNA表达水平;采用流式细胞术检测CD_4~+CD_(25)~+Fox P3细胞、CD_4~+Th1细胞、CD_4~+Th17细胞的比率及炎性反应指标C反应蛋白(CRP)、血沉(ESR)的变化,并比较两组疗效。结果:治疗后观察组总有效率为93.3%(28/30),优于对照组的86.7%(26/30,P<0.05);观察组治疗后CRP、ESR、IL-6、IL-17、IL-23、TNF-α水平及CD_4~+Th17细胞比率较治疗前显著下降(均P<0.05),Fox P3、T-bet的m RNA表达及CD_4~+CD_(25)~+Treg、CD_4~+Th1细胞比率较治疗前显著增加(均P<0.05),且变化程度均较对照组显著(均P<0.05);对照组CRP、ESR、IL-6、IL-17、IL-23、TNF-α水平及CD_4~+Th17细胞比率有所下降,Fox P3、T-bet的mRNA表达及CD_4~+CD_(25)~+Treg、CD_4~+Th1细胞比率有所增加,但治疗前后差异无统计学意义(均P>0.05)。结论:长蛇灸能有效改善AS患者临床症状,调节患者Th17/Treg/Th1免疫失衡,其作用的靶点可能是通过调节AS免疫紊乱及紊乱所致的炎性应答而发挥双向良性调节作用。
Objective To explore the effects and mechanisms of the long-snake moxibustion on ankylosing spondylitis(AS) based on Th17/Treg/Th1 immune imbalance. Methods A total of 60 AS patients were randomized into an observation group and a control group, 30 cases in each one. In the observation group, the long-snake moxibustion therapy was used on the acupoints of the governor vessel from Dazhui(GV 14) to Yaoshu(GV 2) as well as the bilateral Jiaji(EX-B 2) alternatively. The moxibustion was given once a day, for 7 days continuously as one course. There were 3 days at the interval between the courses and 4 courses were required. In the control group, the routine western medication was provided, the salazosulfapyridine combined with non-steroidal anti-inflammatory drugs were used, for 7 days continuous as one course. A total of 4 courses of medication were required. The enzyme linked immunosorbent assay(ELISA) was adopted to determine the levels of interleukin-6(IL-6), interleukin-17(IL-17), interleukin-23(IL-23) and tumor necrosis factor-α(TNF-α). The real-time quantification polymerase chain reaction(RT-PCR) was used to determine the m RNA expressions of the specific transcription factors, Fox P3 and T-bet of the helper 17 cells(Th17), regulatory T cells(Treg) and T helper 1 cells(Th1). The flow cytometry was applied to determine the rates ofCD_4~+CD_(25)~+Treg, CD_4~+Th1 and CD_4~+Th17, as well as the changes of the inflammatory reaction index, C-reactive protein(CRP), and erythrocyte sedimentation rate(ESR). The therapeutic effects were compared between the two groups. Results After treatment, the total effective rate was 93.3%(28/30) in the observation group, which was better than 86.7%(26/30) in the control group(P<0.05). After treatment, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of CD_4~+Th17 were reduced significantly as compared with those before treatment in the observation group(all P<0.05). The mRNA expressions of FoxP3 and T-bet and the rates of CD_4~+CD_(25)~+Treg and CD_4~+Th1 were increased as compared with those before treatment(all P<0.05). The change degree in the observation group was significant as compared with the control group(all P<0.05). In the control group, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of CD_4~+Th17 were reduced, and the mRNA expressions of FoxP3 and T-bet and the rates of CD_4~+CD_(25)~+Treg and CD_4~+Th1 were increased after treatment. But the changes were not significant as compared with those before treatment(all P>0.05). Conclusion The long-snake moxibustion effectively relieves the clinical symptoms in AS patients and regulates the Th17/Treg/Th1 immune imbalance. Its effect target is probably related to the modulation of the AS immune derangement and the inflammatory responses induced by immune derangement so as to achieve the dual-positive regulatory effect.
Objective To explore the effects and mechanisms of the long-snake moxibustion on ankylosing spondylitis(AS) based on Th17/Treg/Th1 immune imbalance. Methods A total of 60 AS patients were randomized into an observation group and a control group, 30 cases in each one. In the observation group, the long-snake moxibustion therapy was used on the acupoints of the governor vessel from Dazhui(GV 14) to Yaoshu(GV 2) as well as the bilateral Jiaji(EX-B 2) alternatively. The moxibustion was given once a day, for 7 days continuously as one course. There were 3 days at the interval between the courses and 4 courses were required. In the control group, the routine western medication was provided, the salazosulfapyridine combined with non-steroidal anti-inflammatory drugs were used, for 7 days continuous as one course. A total of 4 courses of medication were required. The enzyme linked immunosorbent assay(ELISA) was adopted to determine the levels of interleukin-6(IL-6), interleukin-17(IL-17), interleukin-23(IL-23) and tumor necrosis factor-α(TNF-α). The real-time quantification polymerase chain reaction(RT-PCR) was used to determine the m RNA expressions of the specific transcription factors, Fox P3 and T-bet of the helper 17 cells(Th17), regulatory T cells(Treg) and T helper 1 cells(Th1). The flow cytometry was applied to determine the rates ofCD_4~+CD_(25)~+Treg, CD_4~+Th1 and CD_4~+Th17, as well as the changes of the inflammatory reaction index, C-reactive protein(CRP), and erythrocyte sedimentation rate(ESR). The therapeutic effects were compared between the two groups. Results After treatment, the total effective rate was 93.3%(28/30) in the observation group, which was better than 86.7%(26/30) in the control group(P<0.05). After treatment, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of CD_4~+Th17 were reduced significantly as compared with those before treatment in the observation group(all P<0.05). The mRNA expressions of FoxP3 and T-bet and the rates of CD_4~+CD_(25)~+Treg and CD_4~+Th1 were increased as compared with those before treatment(all P<0.05). The change degree in the observation group was significant as compared with the control group(all P<0.05). In the control group, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of CD_4~+Th17 were reduced, and the mRNA expressions of FoxP3 and T-bet and the rates of CD_4~+CD_(25)~+Treg and CD_4~+Th1 were increased after treatment. But the changes were not significant as compared with those before treatment(all P>0.05). Conclusion The long-snake moxibustion effectively relieves the clinical symptoms in AS patients and regulates the Th17/Treg/Th1 immune imbalance. Its effect target is probably related to the modulation of the AS immune derangement and the inflammatory responses induced by immune derangement so as to achieve the dual-positive regulatory effect.
引文
[1]王桂叶,孙振昌,张明智.强直性脊柱炎与HLA-B27相关性的分析[J].中原医刊,2006,33(5):11.
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[9]Jandus C,Bioley G,Rivals JP,et al.Increased numbers of circulating polyfunctional Thl7 memory cells in patients with seronegative spondylarthritides[J].Arthritis Rheum,2008,58(8):2307-2317.
[10]Wang X,Lin Z,Wei Q,et al.Expression of IL-23 and IL-17 and effect of IL-23 on l L-17 production in ankylosing spondylitis[J].Rheumatol Int,2009,29(11):1343-1347.
[11]Brown MA.Genetics and the pathogenesis of ankylosing spondylitis[J].Curr Opin Rheumatol,2009,21(4):318-323.
[12]Shen H,Goodall JC,Hill Gaston JS.Frequency and phenotype of peripheral blood Thl7 cells in ankylosing spondylitis and rheumatoid arthritis[J].Arthritis Rheum,2009,60(6):1647-1656.
[13]Zheng Y,Danilenko DM,Valdez P,et al.Interleukin-22,a T(H)17 cytokine,mediates IL-23-induced derma I inflammation and acanthosis[J].Nature,2007,445(7128):648-651.
[14]Tato CM,O’shea JJ.What does it mean to be just 17?[J].Nature,2006,441(11):166-168.
[15]Mangan PR,Harrington LE,Quinn DB,et al.Transfoming growth factor-beta induces development of the Th17 lineage[J].Nature,2006,44l(7090):231-234.
[16]Komiyama YS.Nakae T,Matsuki A,et al.IL-17 plays an important role in the development of experimental autoimmune encephalomyelitis[J].J Immunol,2006,177(1):566-573.
[17]唐照亮,宋小鸽,章复清,等.艾灸抗炎免疫作用机制的实验研究[J].安徽中医学院学报,2003,22(2):31-35.
[18]陈淑增,白剑鹏,谢永华,等.强直性脊柱炎患者外周血Thl7、Treg、Thl免疫失衡及其与疾病活动相关性研究[J].中国免疫学志,2013,29(8):834-847.
[2]Braun J,Bollow M,Remlinger G,et al.Prevalence of spondylarthropathies in HLA-B27 positive and negative blood donors[J].Arthritis Rheum,1998,41(1):58-67.
[3]Zhang N.A resume of epidemiological surveys of several main rheumatic diseases in China[J].Chin Med J(Engl),1998,lll(3):195-196.
[4]Park H,Li Z,Yang XO,et al.A distinct Lineage of CD4 T cells regulate tissue inflammation by producing interleukin 17[J].Nat Immunol,2005,6(11):1133-1141.
[5]左政,姜云武.长蛇灸配合运动干预治疗强直性脊柱炎的临床观察[J].云南中医中药杂志,2012,33(2):15-16.
[6]van der Linden S,Valkenburg HA,Cats A.Evaluation of diagnostic criteria for ankylosing spondylitis:a proposal for modification of the New York criteria[J].Arthritis Rheum,1984,27(4):361-368.
[7]张奉春,黄烽.风湿病学新进展[M].北京:中华医学电子音像出版社,2005:45.
[8]国家中医药管理局.中医病证诊断疗效标准[M].南京:南京大学出版社,1994:30.
[9]Jandus C,Bioley G,Rivals JP,et al.Increased numbers of circulating polyfunctional Thl7 memory cells in patients with seronegative spondylarthritides[J].Arthritis Rheum,2008,58(8):2307-2317.
[10]Wang X,Lin Z,Wei Q,et al.Expression of IL-23 and IL-17 and effect of IL-23 on l L-17 production in ankylosing spondylitis[J].Rheumatol Int,2009,29(11):1343-1347.
[11]Brown MA.Genetics and the pathogenesis of ankylosing spondylitis[J].Curr Opin Rheumatol,2009,21(4):318-323.
[12]Shen H,Goodall JC,Hill Gaston JS.Frequency and phenotype of peripheral blood Thl7 cells in ankylosing spondylitis and rheumatoid arthritis[J].Arthritis Rheum,2009,60(6):1647-1656.
[13]Zheng Y,Danilenko DM,Valdez P,et al.Interleukin-22,a T(H)17 cytokine,mediates IL-23-induced derma I inflammation and acanthosis[J].Nature,2007,445(7128):648-651.
[14]Tato CM,O’shea JJ.What does it mean to be just 17?[J].Nature,2006,441(11):166-168.
[15]Mangan PR,Harrington LE,Quinn DB,et al.Transfoming growth factor-beta induces development of the Th17 lineage[J].Nature,2006,44l(7090):231-234.
[16]Komiyama YS.Nakae T,Matsuki A,et al.IL-17 plays an important role in the development of experimental autoimmune encephalomyelitis[J].J Immunol,2006,177(1):566-573.
[17]唐照亮,宋小鸽,章复清,等.艾灸抗炎免疫作用机制的实验研究[J].安徽中医学院学报,2003,22(2):31-35.
[18]陈淑增,白剑鹏,谢永华,等.强直性脊柱炎患者外周血Thl7、Treg、Thl免疫失衡及其与疾病活动相关性研究[J].中国免疫学志,2013,29(8):834-847.
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