全反式视黄酸预防性给药可减轻类风湿关节炎模型大鼠的炎症
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摘要
目的探索全反式视黄酸(ATRA)对Ⅱ型胶原(CⅡ)诱导的类风湿关节炎模型(CIA)大鼠关节炎形成期关节组织结构、血清中相关炎性细胞因子表达水平,以及软骨损伤相关蛋白表达水平的影响。方法将6~8周龄雌性Wistar大鼠随机分为空白对照组、溶剂对照组、ATRA各剂量组,给予溶剂对照组和ATRA各剂量组大鼠尾部皮内注射CⅡ和不完全弗氏佐剂诱导类风湿关节炎,空白对照组大鼠以相同方式给予等量生理盐水。自初次免疫第2天起,ATRA各剂量组大鼠腹腔注射不同剂量(0.05、0.5、5 mg/kg)的ATRA油剂,溶剂对照组腹腔注射等体积玉米油,空白对照组腹腔注射等体积生理盐水,每周3次,连续3周。观察ATRA对大鼠关节炎指数(AI)评分、膝踝关节组织病理形态学、血清中相关炎性细胞因子的表达水平及软骨损伤相关蛋白表达水平的影响。结果自初次免疫第15天起,ATRA各剂量组和溶剂对照组AI值均显著高于空白对照组(P<0.05),溶剂对照组AI值趋于平稳,ATRA各剂量组AI值呈上升趋势,且低于溶剂对照组(P<0.05);膝踝关节病理切片可见,ATRA各剂量组和溶剂对照组踝关节结构出现严重紊乱,但组间关节损伤的半定量评分差异无统计学意义(P>0.05);此外,与溶剂对照组相比,ATRA各剂量组白介素-17A(IL-17A)和肿瘤坏死因子-α(TNF-α)分泌减少(P<0.05),白介素-10(IL-10)分泌增多(P<0.05);膝关节中解整链蛋白金属蛋白酶(ADAMTS-4)和基质金属蛋白酶(MMP-3)表达下调(P<0.05),其余软骨损伤相关蛋白的表达差异无统计学意义(P>0.05)。结论在胶原诱导关节炎形成期,ATRA可抑制TNF-α、IL-17A等促炎因子的分泌,促进IL-10的分泌,从而减轻CIA大鼠关节炎形成期的炎症反应。说明ATRA在关节炎形成期可延缓疾病的进展,其作用机制可能与纠正Th1/Th2及Th17/Treg失衡有关。
Objective To investigate the effects of prophylactic administration of all- trans retinoic acid(ATRA) in relieving inflammation in a rat model of collagen-induced arthritis(CIA). Methods Female Wistar rats(6 to 8 weeks old) were randomly divided into normal control group, solvent control group, and prophylactic ATRA treatment(0.05, 0.5, and 5 mg/kg) groups.All the rats except for those in normal control group were subjected to subcutaneous injection of type II collagen and incomplete Freund adjuvant in the tails to induce CIA, followed by injection on the following day with saline, corn oil or different doses of ATRA 3 times a week. The arthritis index(AI) scores, histological scores, serum levels of TNF-α, IL-17 A, and IL-10, and expressions of proteases related with cartilage damage were evaluated. Results On the 15 th day after the primary immunization, the AI scores increased significantly in all but the normal control groups; the scores increased progressively in all the 3 ATRA groups but remained lower than that in the solvent control group, which was stable over time. The rats in the 3ATRA groups showed obvious pathologies in the knee and ankle joints, but the semi-quantitative scores of pathology damage showed no significance among them. Compared with those in solvent control group, the serum IL- 17 A and TNF- α levels decreased, serum IL-10 level increased, and the expressions of ADAMT-4 and MMP-3 proteins decreased significantly in the knees in the 3 ATRA groups. Conclusion ATRA can reduce the production of TNF-α and IL-17 A and increase the production of IL- 10 to alleviate the inflammation in rats with CIA. ATRA may delay the progression of RA by correcting the imbalance of Th1/Th2 and Th17/Treg.
Objective To investigate the effects of prophylactic administration of all- trans retinoic acid(ATRA) in relieving inflammation in a rat model of collagen-induced arthritis(CIA). Methods Female Wistar rats(6 to 8 weeks old) were randomly divided into normal control group, solvent control group, and prophylactic ATRA treatment(0.05, 0.5, and 5 mg/kg) groups.All the rats except for those in normal control group were subjected to subcutaneous injection of type II collagen and incomplete Freund adjuvant in the tails to induce CIA, followed by injection on the following day with saline, corn oil or different doses of ATRA 3 times a week. The arthritis index(AI) scores, histological scores, serum levels of TNF-α, IL-17 A, and IL-10, and expressions of proteases related with cartilage damage were evaluated. Results On the 15 th day after the primary immunization, the AI scores increased significantly in all but the normal control groups; the scores increased progressively in all the 3 ATRA groups but remained lower than that in the solvent control group, which was stable over time. The rats in the 3ATRA groups showed obvious pathologies in the knee and ankle joints, but the semi-quantitative scores of pathology damage showed no significance among them. Compared with those in solvent control group, the serum IL- 17 A and TNF- α levels decreased, serum IL-10 level increased, and the expressions of ADAMT-4 and MMP-3 proteins decreased significantly in the knees in the 3 ATRA groups. Conclusion ATRA can reduce the production of TNF-α and IL-17 A and increase the production of IL- 10 to alleviate the inflammation in rats with CIA. ATRA may delay the progression of RA by correcting the imbalance of Th1/Th2 and Th17/Treg.
引文
[1]曾小峰,谭爱春,谢小平,等.我国类风湿关节炎疾病负担和生存质量研究的系统评价[J].中国循证医学杂志,2013,13(3):300-7.
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[5]Zhou X,Hua X,Ding X,et al.Trichostatin differentially regulates Th1 and Th2 responses and alleviates rheumatoid arthritis in mice[J].J Clin Immunol,2011,31(3):395-405.
[6]Mucida D,Pino-Lagos K,Kim G,et al.Retinoic acid can directly promote TGF-beta-mediated Foxp3(+)Treg cell conversion of naive T cells[J].Immunity,2009,30(4):471-2.
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[8]Boissier MC.Cell and cytokine imbalances in rheumatoid synovitis[J].Joint Bone Spine,2011,78(3):230-4.
[9]Miyabe Y,Miyabe C,Nanki T.Could retinoids be a potential treatment for rheumatic diseases[J]?Rheumatol Int,2015,35(1):35-41.
[10]谢雪健,申雨燕,宋杰,等.全反式视黄酸对CIA大鼠血清炎性细胞因子及软骨损伤相关蛋白表达的影响[J].四川大学学报:医学版,2016,47(4):479-84.
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[12]Roth A,Mollenhauer J,Wagner A,et al.Intra-articular injections of high-molecular-weight hyaluronic acid have biphasic effects on joint inflammation and destruction in rat antigen-induced arthritis[J].Arthritis Res Ther,2005,7(3):R677-86.
[13]Holmdahl R,Andersson M,Goldschmidt TJ,et al.Type II Collagen Autoimmunity in Animals and Provocations Leading to Arthritis[J].Immunol Rev,1990,118:193-232.
[14]张敏娜.类风湿关节炎的实验动物模型及其评价[J].实验动物科学,2011,28(4):57-9.
[15]Zhang P,Han D,Tang T,et al.The destruction evaluation in different foot joints:new ideas in collagen-induced arthritis rat model[J].Rheumatol Int,2009,29(6):607-13.
[16]Feldmann M,Brennan FM,Maini RN.Rheumatoid arthritis[J].Cell,1996,85(3):307-10.
[17]Raychaudhuri B,Fisher CJ,Farver CF,et al.Interleukin 10(IL-10)-mediated inhibition of inflammatory cytokine production by human alveolar macrophages[J].Cytokine,2000,12(9):1348-55.
[18]Kimura A,Kishimoto T.IL-6:Regulator of Treg/Th17 balance[J].Eur J Immunol,2010,40(7):1830-5.
[19]Yoshimatsu H,Okazaki F,Ieiri I,et al.Mechaism of the 24 hour rhythm of tumor necrosis factor-alpha formed by onset of rheumatoid arthritis[J].Chronobiol Int,2014,31(4):564-71.
[20]Kwok SK,Park MK,Cho ML,et al.Retinoic acid attenuates rheumatoid inflammation in mice[J].J Immunol,2012,189(2):1062-71.
[21]Nozaki YJ,Yamagata T,Sugiyama M,et al.Anti-inflammatory effect of all-trans-retinoic acid in inflammatory arthritis[J].Clin Immunol,2006,119(3):272-9.
[22]苏金.DDR2调控MMP-13表达的分子机制及其与类风湿性关节炎关节软骨破坏关系的研究[D].第四军医大学,2008.
[23]Glasson SS,Askew R,Sheppard B,et al.Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of osteoarthritis[J].Nature,2005,434(7033):644-8.
[24]杨文芳,周惠琼.蛋白聚糖异常代谢与类风湿关节炎相关性研究进展[J].中华临床医师杂志:电子版,2013,7(10):4485-8.
[25]Yamamoto K,Murphy G,Troeberg L.Extracellular regulation of metalloproteinases[J].Matrix Biol,2015,44-46:255-63.
[2]Mc Innes IB,Schett G.The pathogenesis of rheumatoid arthritis[J].N Engl J Med,2011,365(23):2205-19.
[3]Trentham DE,Townes AS,Kang AH.Autoimmunity to type II collagen:an experimental model of arthritis[J].J Exp,1977,146(3):857-66.
[4]Falgarone G,Semerano L,Rulle S,et al.Targeting lymphocyte activation to treat rheumatoid arthritis[J].Joint Bone Spine,2009,76(4):327-32.
[5]Zhou X,Hua X,Ding X,et al.Trichostatin differentially regulates Th1 and Th2 responses and alleviates rheumatoid arthritis in mice[J].J Clin Immunol,2011,31(3):395-405.
[6]Mucida D,Pino-Lagos K,Kim G,et al.Retinoic acid can directly promote TGF-beta-mediated Foxp3(+)Treg cell conversion of naive T cells[J].Immunity,2009,30(4):471-2.
[7]Pino-Lagos K,Guo Y,Noelle RJ.Retinoic acid:a key player in immunity[J].Biofactors,2010,36(6):430-6.
[8]Boissier MC.Cell and cytokine imbalances in rheumatoid synovitis[J].Joint Bone Spine,2011,78(3):230-4.
[9]Miyabe Y,Miyabe C,Nanki T.Could retinoids be a potential treatment for rheumatic diseases[J]?Rheumatol Int,2015,35(1):35-41.
[10]谢雪健,申雨燕,宋杰,等.全反式视黄酸对CIA大鼠血清炎性细胞因子及软骨损伤相关蛋白表达的影响[J].四川大学学报:医学版,2016,47(4):479-84.
[11]王岩艳.II型胶原诱导的大鼠关节炎动物模型的研究[D].四川大学,2006.
[12]Roth A,Mollenhauer J,Wagner A,et al.Intra-articular injections of high-molecular-weight hyaluronic acid have biphasic effects on joint inflammation and destruction in rat antigen-induced arthritis[J].Arthritis Res Ther,2005,7(3):R677-86.
[13]Holmdahl R,Andersson M,Goldschmidt TJ,et al.Type II Collagen Autoimmunity in Animals and Provocations Leading to Arthritis[J].Immunol Rev,1990,118:193-232.
[14]张敏娜.类风湿关节炎的实验动物模型及其评价[J].实验动物科学,2011,28(4):57-9.
[15]Zhang P,Han D,Tang T,et al.The destruction evaluation in different foot joints:new ideas in collagen-induced arthritis rat model[J].Rheumatol Int,2009,29(6):607-13.
[16]Feldmann M,Brennan FM,Maini RN.Rheumatoid arthritis[J].Cell,1996,85(3):307-10.
[17]Raychaudhuri B,Fisher CJ,Farver CF,et al.Interleukin 10(IL-10)-mediated inhibition of inflammatory cytokine production by human alveolar macrophages[J].Cytokine,2000,12(9):1348-55.
[18]Kimura A,Kishimoto T.IL-6:Regulator of Treg/Th17 balance[J].Eur J Immunol,2010,40(7):1830-5.
[19]Yoshimatsu H,Okazaki F,Ieiri I,et al.Mechaism of the 24 hour rhythm of tumor necrosis factor-alpha formed by onset of rheumatoid arthritis[J].Chronobiol Int,2014,31(4):564-71.
[20]Kwok SK,Park MK,Cho ML,et al.Retinoic acid attenuates rheumatoid inflammation in mice[J].J Immunol,2012,189(2):1062-71.
[21]Nozaki YJ,Yamagata T,Sugiyama M,et al.Anti-inflammatory effect of all-trans-retinoic acid in inflammatory arthritis[J].Clin Immunol,2006,119(3):272-9.
[22]苏金.DDR2调控MMP-13表达的分子机制及其与类风湿性关节炎关节软骨破坏关系的研究[D].第四军医大学,2008.
[23]Glasson SS,Askew R,Sheppard B,et al.Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of osteoarthritis[J].Nature,2005,434(7033):644-8.
[24]杨文芳,周惠琼.蛋白聚糖异常代谢与类风湿关节炎相关性研究进展[J].中华临床医师杂志:电子版,2013,7(10):4485-8.
[25]Yamamoto K,Murphy G,Troeberg L.Extracellular regulation of metalloproteinases[J].Matrix Biol,2015,44-46:255-63.