风湿宁胶囊对类风湿关节炎大鼠Th17/Treg平衡和局部炎性因子的影响
|
摘要
目的:通过观察风湿宁胶囊(FSN)对类风湿关节炎大鼠外周血Th17/Treg平衡和对关节腔滑液中炎性细胞因子的影响,进一步探讨其治疗类风湿关节炎(RA)的作用机制。方法:制备II型胶原诱导性关节炎(CIA)大鼠模型,采用流式细胞学检测方法观察风湿宁胶囊对CIA模型大鼠外周血辅助性T细胞亚群中Th17(IL-17+CD4+)细胞水平和调节性T细胞(CD4+CD25+Treg细胞)平衡状态的影响,并采用流式多重液相芯片技术检测CIA模型大鼠关节腔滑液中炎性细胞因子(TNF-α、IL-6、IL-17A、IL-4、IL-10)的含量,同时与正常大鼠、CIA模型大鼠和雷公藤多苷片(TPT)治疗大鼠比较。结果:与模型组比较,FSN中、高剂量组大鼠Th17细胞比率明显降低(P<0.05),各治疗组调节性T细胞比率有上调趋势,但差异不显著(P>0.05),各治疗组对Th17细胞和Treg细胞的影响与TPT组比较,均无显著性差异(P>0.05);与模型组比较,FSN中、高剂量可明显降低大鼠关节滑液炎症因子TNF-α、IL-6、IL-17A水平(P<0.05),FSN各组对关节滑液IL-4、IL-10水平未见明显的上调作用(P>0.05),与TPT组比较,各治疗组对局部细胞因子IL-6、TNF-α、IL-17A、IL-4、IL-10的影响均无显著性差异(P>0.05)。结论:FSN能够明显降低外周Th17细胞比率,减少关节滑液炎症因子IL-6、TNF-α、IL-17A的分泌,但对外周Treg细胞比率和关节滑液中抗炎相关因子IL-4、IL-10的分泌无明显影响。风湿宁胶囊调节Th17细胞、改善机体免疫失衡状态、抑制滑膜细胞炎症介质过度分泌等作用可能是其治疗RA的重要机制。
This study was aimed to observe the effects of Feng-Shi-Ning(FSN) capsule on the balance of IL-17+CD4+T(Th17) / Foxp3+CD4+CD25+regulatory T(Treg) cells in peripheral blood and the inflammatory media in synovial fluid of rats with rheumatoid arthritis(RA), in order to further study the mechanism of FSN capsule in RA treatment. The collagen-induced arthritis Ⅱ(CIA) was used in the RA rat model establishment. The balance of Th17 and Treg cells in peripheral blood was analyzed by the flow cytometry method for effects of FSN capsule treatment. The levels of TNF-α, IL-6, IL-17 A, IL-4 and IL-10 in synovial fluid were measured by cytometric bead array(CBA) method. The effects of the normal rats, CIA model rats and the rats treated by tripterygium wilfordii polyglycoside tablet(TPT) were compared. The results showed that compared with the model group, the percentage of Th17 cells in both the FSN capsule group and high-dose CIA group were obviously decreased(P < 0.05). The percentage of Treg cells had the tendency of increasing in all treatment groups with no significant difference(P > 0.05). There was no significant difference on Th17 and Treg cells among each treatment group and TPT group(P > 0.05). Compared with the model group, the high-dose and middle dose FSN can obviously reduce the levels of TNF-α, IL-6 and IL-17 A in synovial fluid of rats in each FSN group(P < 0.05). There was no obvious upregulating effect on IL-4 and IL-10 in synovial fluid of each FSN group(P > 0.05). Compared with the TPT group, there were no significant differences on IL-6, TNF-α, IL-17 A, IL-4 and IL-10 in each treatment group(P > 0.05). It was concluded that FSN can obviously reduce the percentage of Th17, decrease the secretion of IL-6, TNF-α and IL-17 A in synovial fluid. However, it had no obvious effects on the percentage of Treg cells, or the secretion of IL-4 and IL-10 in synovial fluid. The mechanism of FSN capsule in RA treatment may be through regulating Th17 cells, adjusting body immune imbalance, and inhibiting the excessive secretion of inflammatory media in synovial fluid.
This study was aimed to observe the effects of Feng-Shi-Ning(FSN) capsule on the balance of IL-17+CD4+T(Th17) / Foxp3+CD4+CD25+regulatory T(Treg) cells in peripheral blood and the inflammatory media in synovial fluid of rats with rheumatoid arthritis(RA), in order to further study the mechanism of FSN capsule in RA treatment. The collagen-induced arthritis Ⅱ(CIA) was used in the RA rat model establishment. The balance of Th17 and Treg cells in peripheral blood was analyzed by the flow cytometry method for effects of FSN capsule treatment. The levels of TNF-α, IL-6, IL-17 A, IL-4 and IL-10 in synovial fluid were measured by cytometric bead array(CBA) method. The effects of the normal rats, CIA model rats and the rats treated by tripterygium wilfordii polyglycoside tablet(TPT) were compared. The results showed that compared with the model group, the percentage of Th17 cells in both the FSN capsule group and high-dose CIA group were obviously decreased(P < 0.05). The percentage of Treg cells had the tendency of increasing in all treatment groups with no significant difference(P > 0.05). There was no significant difference on Th17 and Treg cells among each treatment group and TPT group(P > 0.05). Compared with the model group, the high-dose and middle dose FSN can obviously reduce the levels of TNF-α, IL-6 and IL-17 A in synovial fluid of rats in each FSN group(P < 0.05). There was no obvious upregulating effect on IL-4 and IL-10 in synovial fluid of each FSN group(P > 0.05). Compared with the TPT group, there were no significant differences on IL-6, TNF-α, IL-17 A, IL-4 and IL-10 in each treatment group(P > 0.05). It was concluded that FSN can obviously reduce the percentage of Th17, decrease the secretion of IL-6, TNF-α and IL-17 A in synovial fluid. However, it had no obvious effects on the percentage of Treg cells, or the secretion of IL-4 and IL-10 in synovial fluid. The mechanism of FSN capsule in RA treatment may be through regulating Th17 cells, adjusting body immune imbalance, and inhibiting the excessive secretion of inflammatory media in synovial fluid.
引文
1周萍,陈欧娜,苏咏梅,等.温经散寒通络汤联合针灸治疗类风湿性关节炎寒湿痹的临床研究.世界科学技术-中医药现代化,2014,16(4):784-789.
2 唐志宇,梁江.昆山合剂对类风湿关节炎合并贫血患者滑膜成纤维细胞增殖及My D88、IL-6表达的研究.世界科学技术-中医药现代化,2014,16(3):582-586.
3 陈俊伟,张少然,闫成兰,等.Th17/调节性T细胞在类风湿关节炎中的作用及其意义.中华风湿病学杂志,2012,16(4):229-232.
4 杨金娜,刘晓光,李覃,等.Th17/Treg平衡在类风湿关节炎中作用的研究进展.中国药理学通报,2013,29(8):1045-1048.
5 徐叔云,卞如濂,陈修.药理实验方法学.北京:人民卫生出版社,2006:1426-1428.
6 王天,赵义,刘栩,等.Th17细胞/调节性T细胞偏倚在类风湿关节炎中的意义.中华风湿病学杂志,2009,13(7):439-442.
7 段一娜,王明娟,杨佳琪,等.穿山龙水溶性总皂苷对RA细胞模型及其下游因子的影响.世界科学技术-中医药现代化,2014,16(6):1396-1400.
8 黄颖.中医对类风湿性关节炎病因病机的认识.辽宁中医药大学学报,2007,9(4):36-37.
9 史丽璞,和雅,郇稳,等.联合用药对类风湿关节炎促炎症细胞因子与急性相反应物的影响及疗效分析.实用医院临床杂志,2013,10(3):78-79.
10 李旭光,张静,李世梅.类风湿关节炎患者血清IL-6、TNF-α水平及其意义.中国现代实用医学杂志,2006,5(9):50-51.
11 梁秀云.类风湿关节炎血清IL-6、TNF-α、CRP的水平变化及意义.海南医学,20l2,23(4):85-87.
12 谭刘欣,奚永志.CD4+CD25+Treg细胞在类风湿关节炎中的研究进展.中华医学杂志,2008,88(27):1937-1939.
13 王庆保,刘健,万磊.类风湿关节炎患者外周血CD4+CD25+CD127-调节T细胞的检测及临床意义.安徽医学,2010,31(2):131-134.
14 牛倩,黄卓春,蔡蓓,等.类风湿性关节炎患者外周血Th17/Treg细胞比率失衡的研究.细胞与分子免疫学杂志,2010,26(3):267-269,272.
15 史浩,朱平.Th17细胞及相关细胞因子在类风湿关节炎中的检测及临床意义.中国现代医生,2013,51(11):101-102,105.
16 Miossec P.Interleukin-17 in fashion,at last:ten years after its description,its cellular source has been identified.Arthritis Rheum,2007,56(7):2111-2115.
17 姚寿林,徐建华.Th17细胞/调节性T细胞平衡与类风湿关节炎.中华风湿病学杂志,2010,14(12):835-837.
18 郑东海,李晶,吴玲,等.来氟米特对类风湿关节炎患者Th17细胞免疫调节的影响.江苏大学学报(医学版),2012,22(1):24-26,30.
19 张雯丽,李慧,闫伟伟.类风湿关节炎患者血清IL-17、TNF-α检测的意义.解放军医药杂志,2011,23(5):l6-18.
20 姚寿林,徐建华,连莉,等.类风湿关节炎患者外周血Th17细胞/调节性T细胞变化及其临床意义.中华风湿病学杂志,2011,15(5):309-313.
21 马艳苗,李艳彦,王永辉,等.风湿宁胶囊对活性氧介导的T细胞活化和滑膜凋亡的影响.中国中西医结合杂志,2013,33(11):1552-1556.
2 唐志宇,梁江.昆山合剂对类风湿关节炎合并贫血患者滑膜成纤维细胞增殖及My D88、IL-6表达的研究.世界科学技术-中医药现代化,2014,16(3):582-586.
3 陈俊伟,张少然,闫成兰,等.Th17/调节性T细胞在类风湿关节炎中的作用及其意义.中华风湿病学杂志,2012,16(4):229-232.
4 杨金娜,刘晓光,李覃,等.Th17/Treg平衡在类风湿关节炎中作用的研究进展.中国药理学通报,2013,29(8):1045-1048.
5 徐叔云,卞如濂,陈修.药理实验方法学.北京:人民卫生出版社,2006:1426-1428.
6 王天,赵义,刘栩,等.Th17细胞/调节性T细胞偏倚在类风湿关节炎中的意义.中华风湿病学杂志,2009,13(7):439-442.
7 段一娜,王明娟,杨佳琪,等.穿山龙水溶性总皂苷对RA细胞模型及其下游因子的影响.世界科学技术-中医药现代化,2014,16(6):1396-1400.
8 黄颖.中医对类风湿性关节炎病因病机的认识.辽宁中医药大学学报,2007,9(4):36-37.
9 史丽璞,和雅,郇稳,等.联合用药对类风湿关节炎促炎症细胞因子与急性相反应物的影响及疗效分析.实用医院临床杂志,2013,10(3):78-79.
10 李旭光,张静,李世梅.类风湿关节炎患者血清IL-6、TNF-α水平及其意义.中国现代实用医学杂志,2006,5(9):50-51.
11 梁秀云.类风湿关节炎血清IL-6、TNF-α、CRP的水平变化及意义.海南医学,20l2,23(4):85-87.
12 谭刘欣,奚永志.CD4+CD25+Treg细胞在类风湿关节炎中的研究进展.中华医学杂志,2008,88(27):1937-1939.
13 王庆保,刘健,万磊.类风湿关节炎患者外周血CD4+CD25+CD127-调节T细胞的检测及临床意义.安徽医学,2010,31(2):131-134.
14 牛倩,黄卓春,蔡蓓,等.类风湿性关节炎患者外周血Th17/Treg细胞比率失衡的研究.细胞与分子免疫学杂志,2010,26(3):267-269,272.
15 史浩,朱平.Th17细胞及相关细胞因子在类风湿关节炎中的检测及临床意义.中国现代医生,2013,51(11):101-102,105.
16 Miossec P.Interleukin-17 in fashion,at last:ten years after its description,its cellular source has been identified.Arthritis Rheum,2007,56(7):2111-2115.
17 姚寿林,徐建华.Th17细胞/调节性T细胞平衡与类风湿关节炎.中华风湿病学杂志,2010,14(12):835-837.
18 郑东海,李晶,吴玲,等.来氟米特对类风湿关节炎患者Th17细胞免疫调节的影响.江苏大学学报(医学版),2012,22(1):24-26,30.
19 张雯丽,李慧,闫伟伟.类风湿关节炎患者血清IL-17、TNF-α检测的意义.解放军医药杂志,2011,23(5):l6-18.
20 姚寿林,徐建华,连莉,等.类风湿关节炎患者外周血Th17细胞/调节性T细胞变化及其临床意义.中华风湿病学杂志,2011,15(5):309-313.
21 马艳苗,李艳彦,王永辉,等.风湿宁胶囊对活性氧介导的T细胞活化和滑膜凋亡的影响.中国中西医结合杂志,2013,33(11):1552-1556.