风湿祛痛胶囊对大鼠胶原诱导性关节炎的影响
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摘要
目的:观察风湿祛痛胶囊对大鼠胶原诱导性关节炎(CIA)的干预作用。方法:SD大鼠随机分为正常组、模型组、风湿祛痛胶囊低、中、高剂量组(0. 25,0. 5,1 g·kg~(-1)·d~(-1)),对照药甲氨蝶呤组(0. 2 mg·kg~(-1)·d~(-1)),除正常组外,其他各组以Ⅱ型胶原和不完全弗氏佐剂免疫大鼠建立CIA模型,首次免疫后第1天给药组开始灌胃给药,每天1次,共给药19 d;首次免疫第8天开始观察大鼠关节红肿和畸形症状、评价关节炎临床积分和发病率,第19天取材后,炎症关节进行微计算机断层扫描技术检测和骨计量学分析,同时进行炎症关节组织病理学检查并从滑膜炎症、血管翳、软骨侵蚀和骨破坏等方面进行半定量评分,酶联免疫吸附测定法检测血清白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α),血管内皮生长因子(VEGF),基质金属蛋白酶-3(MMP-3)和核转录因子-κB受体活化因子配基(RANKL)含量。结果:风湿祛痛胶囊能剂量依赖地改善CIA大鼠炎症关节红、肿和畸形症状;与正常组比较,模型组大鼠炎症关节红、肿和畸形明显临床积分和发病率、关节滑膜炎症、血管翳、软骨侵蚀和骨破坏的病理评分显著升高(P <0. 01),炎症关节骨密度(BMD),骨体积分数(BV/TV),骨小梁厚度(Tb. Th),骨小梁数量(Tb. N)显著降低(P <0. 01),骨表面积体积比(BS/BV)和骨小梁分离度(Tb. Sp)显著升高(P <0. 01),血清IL-1β,TNF-α,VEGF,MMP3和RANKL的含量显著升高(P <0. 01);与模型组相比,各风湿祛痛胶囊组红肿、畸形症状明显改善,临床积分和发病率、关节滑膜炎症、血管翳、软骨侵蚀和骨破坏的病理评分均明显降低(P <0. 05,P <0. 01),BMD,BV/TV,Tb.Th,Tb. N明显升高(P <0. 05,P <0. 01),BS/BV和Tb. Sp明显降低(P <0. 05,P <0. 01),血清IL-1β,TNF-α,VEGF,MMP-3和RANKL的含量明显降低(P <0. 05,P <0. 01),且甲氨蝶呤组与高剂量组的作用相近。结论:风湿祛痛胶囊能有效缓解CIA的临床症状和病情,降低发病率,减轻关节的组织病理学和影像学严重度,同时抑制血清中的促炎细胞因子含量。
Objective: To observe the intervention effect of Fengshi Qutong capsule on collagen-induced arthritis( CIA) in rats. Method: SD rats were randomly divided into normal group,model group,low,medium and high-dose Fengshi Qutong capsule groups( 0. 25, 0. 5, 1 g ·kg~(-1)·d~(-1)), and methotrexate group( 0. 2 mg·kg~(-1)·d~(-1)). Except for normal group,the other groups were immunized with type Ⅱ collagen and incomplete Freund's adjuvant to establish a CIA model. On the 1 stday after the first immunization, the administration group was given intragastric administration,once a day,for 19 days; on the 8 thday after the first immunization,the symptoms of joint swelling and malformation of the rats were observed,and the clinical scores and incidence of arthritis were evaluated. On the 19 thday,micro-computed tomography and bone metrology were performed,and histopathological examination of inflammatory joints was performed, andsynovial inflammation,vasospasm,cartilage erosion and bone destruction by pathological severity scores,serum interleukin-1β( IL-1β),tumor necrosis factor-α( TNF-α), vascular endothelial growth factor( VEGF), matrix metalloproteinase-3( MMP-3) and receptor activator of nuclear factor kappa B ligand( RANKL) were detected by enzyme linked immunosorbent assay. Result: Fengshi Qutong capsule could improve the symptoms of inflammatory joint redness,swelling and deformity in CIA rats in a dose-dependent manner. Compared with normal group,clinical score and incidence,joint synovial inflammation,vasospasm,cartilage erosion and pathological score of bone destruction in joint group were significantly increased( P < 0. 01),bone mineral density( BMD),bone volume/total volume( BV/TV),trabecular thickness( Tb. Th),trabecularnumber( Tb. N) were significantly decreased( P < 0. 01),bone surface area/bone volume( BS/BV) and trabecular separation( Tb. Sp) were significantly elevated( P < 0. 01),and levels of IL-1β,TNF-α,VEGF,MMP-3 and RANKL in serum were increased( P < 0. 01); compared with model group,clinical scores and incidence,joint synovial inflammation,vasospasm,cartilage erosion and bone destruction of each drug groupwere significantly lower( P < 0. 05,P < 0. 01),BMD,BV/TV,Tb. Th,Tb. N were significantly elevated( P < 0. 05,P < 0. 01),BS/BV and Tb. Sp were significantly decreased( P < 0. 05,P < 0. 01),and levels of serum IL-1β,TNF-α,VEGF,MMP-3 and RANKL were significantly decreased( P < 0. 05,P < 0. 01). And the control drug methotrexate had similar effects to the high-dose group. Conclusion: Fengshi Qutong capsule can effectively alleviate the clinical symptoms and conditions of experimental rheumatoid arthritis in rats,reduce the incidence,and relieve the histopathology and imaging severity,while inhibiting the inflammatory cytokines.
Objective: To observe the intervention effect of Fengshi Qutong capsule on collagen-induced arthritis( CIA) in rats. Method: SD rats were randomly divided into normal group,model group,low,medium and high-dose Fengshi Qutong capsule groups( 0. 25, 0. 5, 1 g ·kg~(-1)·d~(-1)), and methotrexate group( 0. 2 mg·kg~(-1)·d~(-1)). Except for normal group,the other groups were immunized with type Ⅱ collagen and incomplete Freund's adjuvant to establish a CIA model. On the 1 stday after the first immunization, the administration group was given intragastric administration,once a day,for 19 days; on the 8 thday after the first immunization,the symptoms of joint swelling and malformation of the rats were observed,and the clinical scores and incidence of arthritis were evaluated. On the 19 thday,micro-computed tomography and bone metrology were performed,and histopathological examination of inflammatory joints was performed, andsynovial inflammation,vasospasm,cartilage erosion and bone destruction by pathological severity scores,serum interleukin-1β( IL-1β),tumor necrosis factor-α( TNF-α), vascular endothelial growth factor( VEGF), matrix metalloproteinase-3( MMP-3) and receptor activator of nuclear factor kappa B ligand( RANKL) were detected by enzyme linked immunosorbent assay. Result: Fengshi Qutong capsule could improve the symptoms of inflammatory joint redness,swelling and deformity in CIA rats in a dose-dependent manner. Compared with normal group,clinical score and incidence,joint synovial inflammation,vasospasm,cartilage erosion and pathological score of bone destruction in joint group were significantly increased( P < 0. 01),bone mineral density( BMD),bone volume/total volume( BV/TV),trabecular thickness( Tb. Th),trabecularnumber( Tb. N) were significantly decreased( P < 0. 01),bone surface area/bone volume( BS/BV) and trabecular separation( Tb. Sp) were significantly elevated( P < 0. 01),and levels of IL-1β,TNF-α,VEGF,MMP-3 and RANKL in serum were increased( P < 0. 01); compared with model group,clinical scores and incidence,joint synovial inflammation,vasospasm,cartilage erosion and bone destruction of each drug groupwere significantly lower( P < 0. 05,P < 0. 01),BMD,BV/TV,Tb. Th,Tb. N were significantly elevated( P < 0. 05,P < 0. 01),BS/BV and Tb. Sp were significantly decreased( P < 0. 05,P < 0. 01),and levels of serum IL-1β,TNF-α,VEGF,MMP-3 and RANKL were significantly decreased( P < 0. 05,P < 0. 01). And the control drug methotrexate had similar effects to the high-dose group. Conclusion: Fengshi Qutong capsule can effectively alleviate the clinical symptoms and conditions of experimental rheumatoid arthritis in rats,reduce the incidence,and relieve the histopathology and imaging severity,while inhibiting the inflammatory cytokines.
引文
[1]郭炜,刘春芳,林娜.类风湿性关节炎滑膜血管新生与中医药[J].中国实验方剂学杂志,2012,18(10):308-312.
[2]张春华.甲氨蝶呤与来氟米特治疗类风湿性关节炎的临床疗效和安全性研究[J].重庆医学,2015,44(4):511-513.
[3]姚晖.风湿祛痛胶囊结合穴位注射治疗类风湿性关节炎的临床研究[J].世界中医药,2016,11(1):52-54.
[4]孙璠璠,李萍,朱志杰,等.风湿祛痛胶囊对大鼠佐剂性关节炎的治疗作用及机制[J].世界中医药,2015,10(12):1918-1921,1925.
[5] Bevaart L,Vervoordeldonk M J,Tak P P. Evaluation of therapeutic targets in animal models of arthritis:how does it relate to rheumatoid arthritis?[J]. Arthritis Rheum,2010,62(8):2192-2205.
[6] LIU C,YANG Y,SUN D,et al. Total saponin from Anemone flaccida Fr. schmidt prevents bone destruction in experimental rheumatoid arthritis via inhibiting osteoclastogenesis[J]. Rejuvenation Res,2015,18(6):528-542.
[7] Remmers E F,Joe B,Griffiths M M,et al. Modulation of multiple experimental arthritis models by collageninducedarthritis quantitative trait loci isolated in congenic rat lines:different effects of non-major histocompatibility complex quantitative trait loci in males and females[J]. Arthritis Rheum,2002,46(8):2225-2234.
[8] ZHANG Y,BAI M,ZHANG B, et al. Uncovering pharmacological mechanisms of wu-tou decoction acting on rheumatoid arthritis through systems approaches:drug-target prediction, network analysis and experimental validation[J]. Sci Rep,2015,5:9463.
[9] Bendele A,Mcabee T,Sennello G,et al. Efficacy of sustained blood levels of interleukin-1 receptor antagonist in animal models of arthritis:comparison of efficacy in animal models with human clinical data[J].Arthritis Rheum,1999,42(3):498-506.
[10]李培培,解国雄,宋珊珊,等.大鼠佐剂性关节炎模型表现特征及评价指标[J].中国免疫学杂志,2012,28(5):453-457.
[11] Hegen M,Keith J C,Collins M,et al. Utility of animal models for identification of potential therapeutics for rheumatoid arthritis[J]. Ann Rheum Dis,2007,67(11):1505-1515.
[12]孙佳蕾,武平,陈白露,等.类风湿关节炎动物模型在中医研究中的应用[J].现代中西医结合杂志,2015,24(4):444-447.
[13]王志中,王勇,牟方祥,等.血小板、TNF-α及IL-1β与活动期类风湿关节炎的相关性研究[J].第三军医大学学报,2011,33(5):469-472.
[14]任海霞,肖诚,李梢,等.类风湿性关节炎中的血管生成相关因子研究进展[J].中国中医基础医学杂志,2007(4):315-318.
[15]赵志玲.基质金属蛋白酶及其抑制剂与类风湿性关节炎的研究进展[J].解放军药学学报,2005(1):54-56.
[16] Okada Y, Nagase H, Harris E D. Matrix metalloproteinases1,2,and 3 from rheumatoid synovial cells are sufficient to destroy joints[J]. J Rheumatol,1987:41-42.
[17] Wada T,Nakashima T,Hiroshi N,et al. Rankl-rank signaling in osteoclastogenesis and bone disease[J].Trends Mol Med,2006,12(1):17-25.
[2]张春华.甲氨蝶呤与来氟米特治疗类风湿性关节炎的临床疗效和安全性研究[J].重庆医学,2015,44(4):511-513.
[3]姚晖.风湿祛痛胶囊结合穴位注射治疗类风湿性关节炎的临床研究[J].世界中医药,2016,11(1):52-54.
[4]孙璠璠,李萍,朱志杰,等.风湿祛痛胶囊对大鼠佐剂性关节炎的治疗作用及机制[J].世界中医药,2015,10(12):1918-1921,1925.
[5] Bevaart L,Vervoordeldonk M J,Tak P P. Evaluation of therapeutic targets in animal models of arthritis:how does it relate to rheumatoid arthritis?[J]. Arthritis Rheum,2010,62(8):2192-2205.
[6] LIU C,YANG Y,SUN D,et al. Total saponin from Anemone flaccida Fr. schmidt prevents bone destruction in experimental rheumatoid arthritis via inhibiting osteoclastogenesis[J]. Rejuvenation Res,2015,18(6):528-542.
[7] Remmers E F,Joe B,Griffiths M M,et al. Modulation of multiple experimental arthritis models by collageninducedarthritis quantitative trait loci isolated in congenic rat lines:different effects of non-major histocompatibility complex quantitative trait loci in males and females[J]. Arthritis Rheum,2002,46(8):2225-2234.
[8] ZHANG Y,BAI M,ZHANG B, et al. Uncovering pharmacological mechanisms of wu-tou decoction acting on rheumatoid arthritis through systems approaches:drug-target prediction, network analysis and experimental validation[J]. Sci Rep,2015,5:9463.
[9] Bendele A,Mcabee T,Sennello G,et al. Efficacy of sustained blood levels of interleukin-1 receptor antagonist in animal models of arthritis:comparison of efficacy in animal models with human clinical data[J].Arthritis Rheum,1999,42(3):498-506.
[10]李培培,解国雄,宋珊珊,等.大鼠佐剂性关节炎模型表现特征及评价指标[J].中国免疫学杂志,2012,28(5):453-457.
[11] Hegen M,Keith J C,Collins M,et al. Utility of animal models for identification of potential therapeutics for rheumatoid arthritis[J]. Ann Rheum Dis,2007,67(11):1505-1515.
[12]孙佳蕾,武平,陈白露,等.类风湿关节炎动物模型在中医研究中的应用[J].现代中西医结合杂志,2015,24(4):444-447.
[13]王志中,王勇,牟方祥,等.血小板、TNF-α及IL-1β与活动期类风湿关节炎的相关性研究[J].第三军医大学学报,2011,33(5):469-472.
[14]任海霞,肖诚,李梢,等.类风湿性关节炎中的血管生成相关因子研究进展[J].中国中医基础医学杂志,2007(4):315-318.
[15]赵志玲.基质金属蛋白酶及其抑制剂与类风湿性关节炎的研究进展[J].解放军药学学报,2005(1):54-56.
[16] Okada Y, Nagase H, Harris E D. Matrix metalloproteinases1,2,and 3 from rheumatoid synovial cells are sufficient to destroy joints[J]. J Rheumatol,1987:41-42.
[17] Wada T,Nakashima T,Hiroshi N,et al. Rankl-rank signaling in osteoclastogenesis and bone disease[J].Trends Mol Med,2006,12(1):17-25.