新风胶囊通过调节氧化应激介导NF-κB信号通路改善佐剂性关节炎大鼠肺功能
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摘要
目的:观察新风胶囊对佐剂性关节炎(AA)大鼠血清细胞因子、氧化应激指标、肺组织中蛋白激酶C(PKC)、核转录因子kappa B(NF-κB)的影响,探讨其防治类风湿关节炎的可能机制。方法:40只雄性SD大鼠随机分为正常组、模型组、新风胶囊组、来氟米特组,每组10只。除正常组外,其余3组均在大鼠右后足趾皮内注射Freund完全佐剂以致炎,诱导AA大鼠模型。致炎后第19天给药,连续30d,1d/次,正常组与模型组用0.9%氯化钠溶液灌胃,新风胶囊组与来氟米特组给予相应剂量的药物。观察各组大鼠足趾肿胀度(E)、关节炎指数(AI)、肺功能,ELISA检测大鼠血清IL-6、IL-17、IL-10、IL-35、NO、iNOS、HIF-1、NADPH的表达,免疫组织化学染色和Western blot法检测大鼠肺组织PKC、NF-κB蛋白表达的水平,荧光定量PCR检测PKC、NF-κB mRNA的表达。结果:与模型组比较,新风胶囊组大鼠SD、AI、肺功能参数、IL-10、IL-35、NADPH、HIF-1表达显著升高(P<0.01,P<0.05),IL-6、IL-17、NO、iNOS表达显著下降(P<0.05,P<0.01),肺组织PKC、NF-κB mRNA和蛋白的表达显著降低(P<0.05,P<0.01)。结论:新风胶囊可能通过调节氧化应激介导的NF-κB通路,改善肺功能。
Objective: To observe the effect of Xinfeng Capsule(XFC) on serum cytokine, oxidative stress index, protein kinase C(PKC) and nuclear transcription factor kappa B(NF-κB) in lung tissue in the rat model of adjuvant-induced arthritis(AA). Methods: Male SD rats were randomly divided into normal control(NC) group, model control(MC) group, XFC group, and leflunomide group, with 10 rats in each group. Complete Freund's adjuvant(CFA, 0.1 mL) was injected into the right foot plantar skin of all rats except for the NC group. The normal and model rats were orally administered the same volume of saline solution,XFC group and the leflunomide group were given the corresponding doses of the drug, once a day from the 19 th day to the 49 th day after AA induction. The degree of toe swelling(E), arthritis index(AI) and pulmonary function were observed. The expression of IL-6, IL-17, IL-10, IL-35, NO, iNOS, HIF-1, NADPH in rat serum were detected by ELISA. The expression of PKC, NF-κB in rat lung tissue were detected by immunohistochemistry and Western blot. Meanwhile, PKC, NF-κB mRNA expression were determined by reverse transcriptase PCR. Results: Compared with the MC group, SD, AI, the parameters of pulmonary function,IL-10, IL-35, NADPH, HIF-1 expression were significantly increased(P<0.01, P<0.05), IL-6, IL-17, NO, iNOS expression were significantly decreased in the XFC group(P<0.05, P<0.01), while PKC, NF-κB mRNA and protein expression were significantly decreased(P<0.05, P<0.01). Conclusion: XFC may improve the pulmonary function by regulating oxidative stress mediated NF-κB signaling pathway.
Objective: To observe the effect of Xinfeng Capsule(XFC) on serum cytokine, oxidative stress index, protein kinase C(PKC) and nuclear transcription factor kappa B(NF-κB) in lung tissue in the rat model of adjuvant-induced arthritis(AA). Methods: Male SD rats were randomly divided into normal control(NC) group, model control(MC) group, XFC group, and leflunomide group, with 10 rats in each group. Complete Freund's adjuvant(CFA, 0.1 mL) was injected into the right foot plantar skin of all rats except for the NC group. The normal and model rats were orally administered the same volume of saline solution,XFC group and the leflunomide group were given the corresponding doses of the drug, once a day from the 19 th day to the 49 th day after AA induction. The degree of toe swelling(E), arthritis index(AI) and pulmonary function were observed. The expression of IL-6, IL-17, IL-10, IL-35, NO, iNOS, HIF-1, NADPH in rat serum were detected by ELISA. The expression of PKC, NF-κB in rat lung tissue were detected by immunohistochemistry and Western blot. Meanwhile, PKC, NF-κB mRNA expression were determined by reverse transcriptase PCR. Results: Compared with the MC group, SD, AI, the parameters of pulmonary function,IL-10, IL-35, NADPH, HIF-1 expression were significantly increased(P<0.01, P<0.05), IL-6, IL-17, NO, iNOS expression were significantly decreased in the XFC group(P<0.05, P<0.01), while PKC, NF-κB mRNA and protein expression were significantly decreased(P<0.05, P<0.01). Conclusion: XFC may improve the pulmonary function by regulating oxidative stress mediated NF-κB signaling pathway.
引文
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[18]孙玥,刘健,万磊,等.类风湿关节炎患者心肺功能变化与B、T淋巴细胞衰减因子及氧化应激的相关性分析.免疫学杂志,2015,31(3):234-239
[19]王晓玉,孟楣,许坡,等.新风胶囊配伍前后毒性成份雷公藤酯甲的量变研究.中国基层医药,2015,22(4):507-510
[20]Wang Y,Liu J,Wang Y,et al.Effect of Xinfeng capsule in the treatment of active rheumatoid arthritis:A randomized controlled trial.J Tradit Chin Med,2015,25(6):226-231
[21]Wan L,Liu J,Huang C B,et al.Xinfeng capsule for the treatment of rheumatoid arthritis patients with decrease pulmonary fuction-a randomized controlled trial.China Integr Med,2016,22(3):168-176
[22]章平衡,万磊,刘健.基于TGF-β1/Smads和ERK通路cross-talk研究新风胶囊改善佐剂性关节炎大鼠肺功能的机制.中华中医药杂志,2017,32(5):2253-2259
[23]章平衡,刘健,谈冰,等.新风胶囊通过降低NF-κB通路活性改善类风湿关节炎患者血瘀状态.中华中医药杂志,2016,31(11):4684-4689
[2]Kurimoto R,Shono K,Onoda M,et al.MYC/BCL2 double-hit lymphoma in a patient with rheumatoid arthritis associated with methotrexate treatment.Intern Med,2016,5(16):2271-2275
[3]孙玥,刘健,万磊,等.新风胶囊通过调节Keapl-Nrf2/ARE信号通路改善佐剂性关节炎大鼠肺功能研究.中华中医药杂志,2016,31(5):1971-1978
[4]汪君民,赵阶祥.运动热应激对大鼠淋巴细胞凋亡氧化应激机制的影响.辽宁医学院学报,2011,32(2):105-109
[5]Sunanda Kundu,P arashar Ghosh,Suhana Datta,et al.Oxidative stress as a potential biomarker for determining disease activity in patients with Rheumatoid Arthritis.Free Radical Research,2012,46(12):1482-1489
[6]WANG Y,WANG G Z,RANINOVITCH P S,et al.Macrophage mitochondriol oxidative stress promotes atherosclerosis and NF-κB-mediatedinflammationinmacrophage.Circ Res,2014,114(3):421-433
[7]Rahman I,Biswas S K,Jimener LA,at al.Glutathione,stress responses,and redox signaling in lung inflammation.Antioxidant Redox Signal,2005,7:42-59
[8]孙玥,刘健,万磊.新风胶囊对类风湿关节炎患者肺功能的影响.中国中西医结合杂志,2016,36(7):814-820
[9]孙玥,刘健,万磊,等.新风胶囊改善类风湿性关节炎患者心功能的机制.细胞与分子免疫学杂志,2015,31(1):93-96,99
[10]李仪奎.中药药理实验方法学.上海:上海科学技术出版社,1991:305
[11]姬洁莹,李晋奇,郭阿霞.类风湿关节炎动物模型复制与评价.山西中医学院学报,2013,14(3):73-76
[12]徐叔云,卞如濂,陈修.药理实验方法学.北京:人民卫生出版社,1994:719-720
[13]张钧田.现代药理实验方法.北京:北京医科大学,中国协和医科大学联合出版社,1998:1383
[14]Bawadekar M,Andrea M D,Cigno L L,et al.The extracellular IFLL6protein propagaes inflammation in endothelial cells Via p38 MAPK and NF-κB activation.J Interforten Cytokine Res,2015,35:1-13
[15]Choi Y J,Uehara Y,Park J Y,et al.MHY884,a newly synthesized throsinase inhibitor,suppressed UVB-induced activated of NF-κB signaling pathway through the downregulation of oxdative stess.Bioorg Med Chem Lett,2014,24(5):1344-1348
[16]Ma Z,Wang B,Wang M,et al.TL1A increased IL-6 production on fibroblast-like synoviocyles by preferentially activating TNF receptor 2 in rheumatoid arthritis.Cytokine,2016,83:92-98
[17]Wei L,Sun Y,Kong X F,et al.The effects of dopamine receptor 2expression on B cells on bone metabolim and TNF-αlevels in rheumatoid arthritis.BMC Musculoskelet Disord,2016,17:352
[18]孙玥,刘健,万磊,等.类风湿关节炎患者心肺功能变化与B、T淋巴细胞衰减因子及氧化应激的相关性分析.免疫学杂志,2015,31(3):234-239
[19]王晓玉,孟楣,许坡,等.新风胶囊配伍前后毒性成份雷公藤酯甲的量变研究.中国基层医药,2015,22(4):507-510
[20]Wang Y,Liu J,Wang Y,et al.Effect of Xinfeng capsule in the treatment of active rheumatoid arthritis:A randomized controlled trial.J Tradit Chin Med,2015,25(6):226-231
[21]Wan L,Liu J,Huang C B,et al.Xinfeng capsule for the treatment of rheumatoid arthritis patients with decrease pulmonary fuction-a randomized controlled trial.China Integr Med,2016,22(3):168-176
[22]章平衡,万磊,刘健.基于TGF-β1/Smads和ERK通路cross-talk研究新风胶囊改善佐剂性关节炎大鼠肺功能的机制.中华中医药杂志,2017,32(5):2253-2259
[23]章平衡,刘健,谈冰,等.新风胶囊通过降低NF-κB通路活性改善类风湿关节炎患者血瘀状态.中华中医药杂志,2016,31(11):4684-4689