芍甘附子汤加味对CIA寒证大鼠Egr2/Egr3及其信号通路表达的影响
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摘要
目的:研究芍甘附子汤加味对胶原诱导性关节炎(CIA)寒证大鼠Egr2/Egr3及其信号通路表达的影响,并探讨Egr2/Egr3在类风湿关节炎(RA)发病中的作用。方法:110只雌性Wistar大鼠被随机分为7组:正常组、模型组、对照组[甲氨蝶呤(MTX)组、雷公藤组]和芍甘附子汤加味组(大、中、小剂量组)。正常组常规饲养,其余各组在复制CIA寒证大鼠模型后分别MTX、雷公藤多苷片及芍甘附子汤加味(大、中、小剂量)灌胃,干预4周,每周评价各组大鼠炎症指数。4周后测定ELISA法检测大鼠外周血IL-6、TGF-β表达;RT-q PCR检测大鼠膝关节滑膜Egr2/Egr3 m RNA、SOCS1/SOCS3 m RNA、STAT1/STAT3 m RNA表达。实验结束后测定大鼠脏器指数。结果:各组大鼠AI评分均呈下降趋势,但各组间差异无统计学意义。HE染色提示各给药组及模型组存在不同程度的滑膜增生、炎性细胞浸润以及骨侵蚀等,各给药组较模型组均有所减轻,模型组和各给药组脾脏指数显著高于正常组(P<0.05),大、中剂量组胸腺指数较模型组显著下降(P<0.05)。大鼠滑膜组织模型组和各治疗组Egr2/Egr3 m RNA、SOCS1/SOCS3 m RNA相对表达量较正常组升高,其中芍甘附子汤中剂量组Egr2/Egr3 m RNA表现尤为突出(P<0.05)。CIA造模后大鼠外周血TGF-β、IL-6水平均显著高于正常组水平,各药物干预组治疗4周后较模型组均有下降尤以TGF-β更为明显。结论:Egr2/Egr3参与了RA发病过程的调控环节,对于疾病的早期诊断及早期干预有提示作用。
Objective: To study the effect of Shaogan Fuzi Decoction on the expression of Egr2/Egr3 and its signal pathway in collagen-induced arthritis(CIA) rats and to explore the role of Egr2/Egr3 in the pathogenesis of rheumatoid arthritis. Methods: 110 female Wistar rats were randomly divided into 7 groups: normal group, model group, control group(methotrexate(MTX) group, Tripterygium wilfordii group) and Shaogan Fuzi Decoction group( high, medium, low dose). The rats in the control group were treated with MTX, Tripterygium wilfordii tablets. Shaogan Fuzi Decoction group were treated with high, medium and small dose Shaogan Fuzi Decoction, and the rats in each group were treated rats for 4 weeks. Inflammation index of rats in each group was evaluated weekly. After 4 weeks, the expression of IL-6 and TGF-beta in peripheral blood of rats. were detected by ELISA, the expression of Egr2/Egr3 m RNA, SOCS1/SOCS3 m RNA and STAT1/STAT3 m RNA in rat synovium were detected by RT-q PCR. At the end of the experiment, the rat organ index was measured. Results: The scores of AI were decreased in each group, but there was no significant difference between the two groups. HE staining indicated that there were different levels of synovial hyperplasia, inflammatory cell infiltration and bone erosion both in the treated group and the model group, but compared to model group, the inflammation of each treated group were relieved. The spleen index of model group and each treated group was significantly higher than that of normal group, while there was no significant difference among each treated group and model group. The expression of Egr2/Egr3 m RNA and SOCS1/SOCS3 m RNA in the synovial tissue of model group and treated group were higher than those in the normal group. The expression of Egr2/Egr3 m RNA in the middle dose group was significantly higher than that in the control group(P<0.05). The levels of TGF-β and IL-6 in the peripheral blood of rats in model group were significantly higher than those of the normal group, and the levels of TGF-β and IL-6 in the treated group were significantly decreased after 4 weeks of treatment. Conclusion: Egr2/Egr3 is involved in the regulation of RA and plays an important role in the early diagnosis and early intervention of disease.
Objective: To study the effect of Shaogan Fuzi Decoction on the expression of Egr2/Egr3 and its signal pathway in collagen-induced arthritis(CIA) rats and to explore the role of Egr2/Egr3 in the pathogenesis of rheumatoid arthritis. Methods: 110 female Wistar rats were randomly divided into 7 groups: normal group, model group, control group(methotrexate(MTX) group, Tripterygium wilfordii group) and Shaogan Fuzi Decoction group( high, medium, low dose). The rats in the control group were treated with MTX, Tripterygium wilfordii tablets. Shaogan Fuzi Decoction group were treated with high, medium and small dose Shaogan Fuzi Decoction, and the rats in each group were treated rats for 4 weeks. Inflammation index of rats in each group was evaluated weekly. After 4 weeks, the expression of IL-6 and TGF-beta in peripheral blood of rats. were detected by ELISA, the expression of Egr2/Egr3 m RNA, SOCS1/SOCS3 m RNA and STAT1/STAT3 m RNA in rat synovium were detected by RT-q PCR. At the end of the experiment, the rat organ index was measured. Results: The scores of AI were decreased in each group, but there was no significant difference between the two groups. HE staining indicated that there were different levels of synovial hyperplasia, inflammatory cell infiltration and bone erosion both in the treated group and the model group, but compared to model group, the inflammation of each treated group were relieved. The spleen index of model group and each treated group was significantly higher than that of normal group, while there was no significant difference among each treated group and model group. The expression of Egr2/Egr3 m RNA and SOCS1/SOCS3 m RNA in the synovial tissue of model group and treated group were higher than those in the normal group. The expression of Egr2/Egr3 m RNA in the middle dose group was significantly higher than that in the control group(P<0.05). The levels of TGF-β and IL-6 in the peripheral blood of rats in model group were significantly higher than those of the normal group, and the levels of TGF-β and IL-6 in the treated group were significantly decreased after 4 weeks of treatment. Conclusion: Egr2/Egr3 is involved in the regulation of RA and plays an important role in the early diagnosis and early intervention of disease.
引文
[1](美)菲尔斯坦著,栗占国译.凯利风湿病学.9版.北京:北京大学医学出版社,2015:1133
[2]赵新秀,寇永锋.类风湿性关节炎中医证型分布规律的文献研究.中华中医药学刊,2008,26(9):1943-1944
[3]刘小平,袁芳,朱跃兰.温经通络、补脾益肾法治疗虚寒型类风湿关节炎探讨.北京中医药,2015,34(11):881-883
[4]靖卫霞,朱跃兰.芍甘附子汤加味对CIA大鼠关节滑膜组织JAK/STAT通路相关基因表达的影响.中医学报,2016,31(4):545-549
[5]袁芳,朱跃兰.芍甘附子汤加味对胶原诱导性关节炎大鼠滑膜血管生成抑制因子干预作用的机制研究.北京中医药,2014,33(7):551-554
[6]柳洋,朱跃兰,侯秀娟,等.芍甘附子汤加味对胶原诱导性关节炎大鼠滑膜血管新生相关因子表达的影响.环球中医药,2015,8(6):692-696
[7]靖卫霞,侯秀娟,孙文燕,等.温经通络法对CIA大鼠关节滑膜组织VEGF及其受体基因表达的影响.北京中医药,2014,33(3):211-214
[8]袁芳,侯秀娟,刘小平,等.芍甘附子汤加味对Ⅱ型胶原诱导性关节炎大鼠OPG-RANKL表达的影响.中华中医药杂志,2015,30(5):1661-1665
[9]马俊福,侯秀娟,刘小平,等.类风湿关节炎寒证动物模型的建立与评价.中华中医药杂志,2016,31(5):1967-1970
[10]刘栩,贾园,安媛,等.类风湿关节炎患者就医及治疗现状分析.中华风湿病学杂志,2008,9(12):637-640
[11]1opilko P,Schneider-Maunoury S,Levi G,et al.Krox-20 controls myelination in the peripheral nervous system.Nature,1994,371:796-799
[12]Tizong Miao,Meera Raymond,Punamdip Bhullar,et al.Early Growth Response Gene-2 Controls IL-17 Expression and Th17Differentiation.The Journal of Immunity,2013,190:58-65
[13]Tamiya T,Kashiwagi I,Takahashi R,et al.Suppressors of cytokine signaling(SOCS)proteins and JAK/STATpathways:regulation of T-cell inflammation by SOCS1 and SOCS3.Arterioscler.Thromb Vasc Biol,2011,31:980-985
[14]Akira S,Jakeda K.Toll-like receptor signaling.Nat Rev Immunol,2004,4(7):499-511
[15]罗福祥,卢军,张海英,等.新疆阿魏乙酸乙酯部位体内外对CT-26.WT细胞的影响.中成药,2017,39(2):238-243
[16]唐恬,赵婷,赵晖,等.附子及其炮制品对大鼠心脏毒性的影响.中国中医急症,2016,25(12):2286-2289,2315
[2]赵新秀,寇永锋.类风湿性关节炎中医证型分布规律的文献研究.中华中医药学刊,2008,26(9):1943-1944
[3]刘小平,袁芳,朱跃兰.温经通络、补脾益肾法治疗虚寒型类风湿关节炎探讨.北京中医药,2015,34(11):881-883
[4]靖卫霞,朱跃兰.芍甘附子汤加味对CIA大鼠关节滑膜组织JAK/STAT通路相关基因表达的影响.中医学报,2016,31(4):545-549
[5]袁芳,朱跃兰.芍甘附子汤加味对胶原诱导性关节炎大鼠滑膜血管生成抑制因子干预作用的机制研究.北京中医药,2014,33(7):551-554
[6]柳洋,朱跃兰,侯秀娟,等.芍甘附子汤加味对胶原诱导性关节炎大鼠滑膜血管新生相关因子表达的影响.环球中医药,2015,8(6):692-696
[7]靖卫霞,侯秀娟,孙文燕,等.温经通络法对CIA大鼠关节滑膜组织VEGF及其受体基因表达的影响.北京中医药,2014,33(3):211-214
[8]袁芳,侯秀娟,刘小平,等.芍甘附子汤加味对Ⅱ型胶原诱导性关节炎大鼠OPG-RANKL表达的影响.中华中医药杂志,2015,30(5):1661-1665
[9]马俊福,侯秀娟,刘小平,等.类风湿关节炎寒证动物模型的建立与评价.中华中医药杂志,2016,31(5):1967-1970
[10]刘栩,贾园,安媛,等.类风湿关节炎患者就医及治疗现状分析.中华风湿病学杂志,2008,9(12):637-640
[11]1opilko P,Schneider-Maunoury S,Levi G,et al.Krox-20 controls myelination in the peripheral nervous system.Nature,1994,371:796-799
[12]Tizong Miao,Meera Raymond,Punamdip Bhullar,et al.Early Growth Response Gene-2 Controls IL-17 Expression and Th17Differentiation.The Journal of Immunity,2013,190:58-65
[13]Tamiya T,Kashiwagi I,Takahashi R,et al.Suppressors of cytokine signaling(SOCS)proteins and JAK/STATpathways:regulation of T-cell inflammation by SOCS1 and SOCS3.Arterioscler.Thromb Vasc Biol,2011,31:980-985
[14]Akira S,Jakeda K.Toll-like receptor signaling.Nat Rev Immunol,2004,4(7):499-511
[15]罗福祥,卢军,张海英,等.新疆阿魏乙酸乙酯部位体内外对CT-26.WT细胞的影响.中成药,2017,39(2):238-243
[16]唐恬,赵婷,赵晖,等.附子及其炮制品对大鼠心脏毒性的影响.中国中医急症,2016,25(12):2286-2289,2315