补肾抗风湿方药对CIA大鼠骨组织RANKL/RANK/OPG系统的影响
|
摘要
目的:观察补肾抗风湿方药寒痹康汤对CIA大鼠骨组织RANKL/RANK/OPG系统的影响,揭示其抗类风湿关节炎骨质侵蚀的作用机制。方法:采用弗氏不完全佐剂乳化的牛Ⅱ型胶原蛋白诱导CIA大鼠模型,将60只Wistar大鼠随机分为正常对照组,模型对照组,甲氨蝶呤组,寒痹康汤高、中、低剂量组,以甲氨蝶呤组为对照,观察不同剂量寒痹康汤对CIA大鼠骨组织RANKL、RANK和OPG表达水平的影响。结果:与正常对照组比较,模型对照组大鼠骨组织的RANKL m RNA、RANK m RNA、RANKL蛋白、RANK蛋白的表达升高,OPG m RNA、OPG蛋白的表达下降(P<0.01);与模型对照组比较,寒痹康汤各剂量组和甲氨蝶呤组的RANKL m RNA、RANK m RNA、RANKL蛋白、RANK蛋白的表达均有不同程度的下降,OPG m RNA、OPG蛋白的表达均有不同程度的升高(P<0.01或P<0.05),其中以寒痹康汤高剂量组最明显;寒痹康汤高剂量组与甲氨蝶呤组比较,差异无统计学意义(P>0.05)。结论:寒痹康汤能抑制CIA大鼠的关节炎严重度,提高骨组织中OPG蛋白和基因表达,抑制RANKL和RANK的蛋白和基因表达,提示其具有抗类风湿关节炎骨质侵蚀的作用,机制可能与调控OPG/RANKL/RANK破骨细胞分化调节信号传导系统有关。
Objective:To observe the effect of invigorating kidney and anti rheumatism prescription Hanbi Kang Tang(寒痹康汤) on the RANKL/RANK/OPG bone tissue system of CIA rats to reveal its mechanism of anti- Rheumatoid Arthritis bone erosion.Methods:Bovine type II collagen emulsified with Freund's incomplete adjuvant was used to induce CIA rat models.Sixty Wistar rats were randomly divided into a normal control group,a model control group,a methotrexate group,and Hanbi Kang high,medium and low dose groups.The methotrexate group was used as an object of reference to observe the influence of difference doses of Hanbi Kang Tang on the expression of rat bone tissue RANKL,RANK and OPG.Results:Compared with the normal control group,the expressions of RANKL m RNA,RANK m RNA,RANKL,and RANK proteins in rats of the model control group increased,while the expressions of OPG m RNA and OPG proteins decreased(P < 0.01).Compared with the model group,the expressions of RANKL m RNA,RANK m RNA,RANKL,and RANK proteins in rats of the Hanbi Kang groups and the methotrexate group decreased in different degree,while the expression of OPG m RNA and OPG proteins decreased(P < 0.01 or P < 0.05),among which the Hanbi Kang high dose group was the most obvious,with no significant difference compared with the methotrexate group(P > 0.05).Hanbi Kang could inhibit the progression of arthritis in CIA rats,improving the expressions of OPG protein and gene in bone tissue,and inhibiting the expressions of RANKL and RANK proteins and gene.Conclusion:Hanbi Kang Tang has the effect of anti-RA bone erosion,whose mechanism may be related to the regulation of OPG/RANKL/RANK osteoclast differentiation regulating signal transduction system.
Objective:To observe the effect of invigorating kidney and anti rheumatism prescription Hanbi Kang Tang(寒痹康汤) on the RANKL/RANK/OPG bone tissue system of CIA rats to reveal its mechanism of anti- Rheumatoid Arthritis bone erosion.Methods:Bovine type II collagen emulsified with Freund's incomplete adjuvant was used to induce CIA rat models.Sixty Wistar rats were randomly divided into a normal control group,a model control group,a methotrexate group,and Hanbi Kang high,medium and low dose groups.The methotrexate group was used as an object of reference to observe the influence of difference doses of Hanbi Kang Tang on the expression of rat bone tissue RANKL,RANK and OPG.Results:Compared with the normal control group,the expressions of RANKL m RNA,RANK m RNA,RANKL,and RANK proteins in rats of the model control group increased,while the expressions of OPG m RNA and OPG proteins decreased(P < 0.01).Compared with the model group,the expressions of RANKL m RNA,RANK m RNA,RANKL,and RANK proteins in rats of the Hanbi Kang groups and the methotrexate group decreased in different degree,while the expression of OPG m RNA and OPG proteins decreased(P < 0.01 or P < 0.05),among which the Hanbi Kang high dose group was the most obvious,with no significant difference compared with the methotrexate group(P > 0.05).Hanbi Kang could inhibit the progression of arthritis in CIA rats,improving the expressions of OPG protein and gene in bone tissue,and inhibiting the expressions of RANKL and RANK proteins and gene.Conclusion:Hanbi Kang Tang has the effect of anti-RA bone erosion,whose mechanism may be related to the regulation of OPG/RANKL/RANK osteoclast differentiation regulating signal transduction system.
引文
[1]邓安梅,仲人前,陈孙孝,等.关节炎中B7:CD28/CTLA-4共刺激途径的实验研究[J].中国免疫学杂志,2000,16(8):412-414.
[2]徐淑云,卞如濂,陈修.药理学实验方法学[M].3版.北京:人民卫生出版社,2002:1861.
[3]Abu-Amer Y,Abbas S,Hirayama T.TNF receptor typeI regulates RANK ligand espression by stromal cellsand modulates osteoclastogenesis[J].J Cell Biochem,2004,93(5):980-989.
[4]Saidenberg-Kermanac'h N,Conado A,Lemeiter D.TNF-αantibodies and osteoprotegerin decrease systemic bone loss associated with inflammation throughdistinct mechanisms in collagen-induced arthritis[J].Bone,2004,35(5):1200-1207.
[5]Page G,Miossec P.RANK and RANKL expression asmarkers of dendritic cell-T cel1 interactions in pairedsamples of rheumatoid synovium and lymph nodes[J].Arthritis Rheum,2005,52(8):2307-2312.
[6]Jones DH,Kong YY,Penninger JM.Role of RANKLand RANK in bone loss and arthritis[J].Ann RheumDis,2002,61 Suppl2:32-39.
[7]Romas E,Gillespie MT,Martin TJ.Involvement of receptoractivator of NFkappa B ligand and tumor necrosisfactor-αin bone destruction in rheumatoid arthritis[J].Bone,2002,30(2):340-346.
[8]Coetzee M,Kruger MC.Osteoprotegerin-receptor activator of nuclear factor-kappa B ligand ratio:a newapproach to osteoporosis treatment?[J].South MedJ,2004,97(5):506-511.
[9]Pettit AR,Walsh NC,Manning C,et al.RANKL proteinisexpressed at the pannus-bone interface at sites of articular bone erosion in rheumatoid arthritis[J].Rheumatology(Oxford),2006,45(9):1068-1076.
[10]庞学丰,吴燕红,蒙宇华.补益肝肾、祛湿蠲痹法治疗寒湿型类风湿关节炎的临床研究[J].中国中医骨伤科杂志,2006,14(1):40-42.
[11]庞学丰,刘欢,吴燕红,等.补肾抗风湿中药联合西药治疗类风湿关节炎继发骨质疏松临床研究[J].世界中西医结合杂志,2015,10(1):47-49.
[12]庞学丰,任志宏,程世和,等.寒痹康汤对实验性关节炎大鼠血清IL-1β及IL-4的影响[J].陕西中医,2009,30(7):919-920.
[13]庞学丰,王乾,吴燕红,等.寒痹康汤对胶原诱导性关节炎大鼠滑膜细胞NF-κB表达的影响[J].风湿病与关节炎,2013,2(7):32-34.
[14]庞学丰,舒建龙,李玉玲,等.寒痹康汤对胶原诱导性关节炎大鼠血清VEGF的影响[J].陕西中医,2015,36(5):616-618.
[2]徐淑云,卞如濂,陈修.药理学实验方法学[M].3版.北京:人民卫生出版社,2002:1861.
[3]Abu-Amer Y,Abbas S,Hirayama T.TNF receptor typeI regulates RANK ligand espression by stromal cellsand modulates osteoclastogenesis[J].J Cell Biochem,2004,93(5):980-989.
[4]Saidenberg-Kermanac'h N,Conado A,Lemeiter D.TNF-αantibodies and osteoprotegerin decrease systemic bone loss associated with inflammation throughdistinct mechanisms in collagen-induced arthritis[J].Bone,2004,35(5):1200-1207.
[5]Page G,Miossec P.RANK and RANKL expression asmarkers of dendritic cell-T cel1 interactions in pairedsamples of rheumatoid synovium and lymph nodes[J].Arthritis Rheum,2005,52(8):2307-2312.
[6]Jones DH,Kong YY,Penninger JM.Role of RANKLand RANK in bone loss and arthritis[J].Ann RheumDis,2002,61 Suppl2:32-39.
[7]Romas E,Gillespie MT,Martin TJ.Involvement of receptoractivator of NFkappa B ligand and tumor necrosisfactor-αin bone destruction in rheumatoid arthritis[J].Bone,2002,30(2):340-346.
[8]Coetzee M,Kruger MC.Osteoprotegerin-receptor activator of nuclear factor-kappa B ligand ratio:a newapproach to osteoporosis treatment?[J].South MedJ,2004,97(5):506-511.
[9]Pettit AR,Walsh NC,Manning C,et al.RANKL proteinisexpressed at the pannus-bone interface at sites of articular bone erosion in rheumatoid arthritis[J].Rheumatology(Oxford),2006,45(9):1068-1076.
[10]庞学丰,吴燕红,蒙宇华.补益肝肾、祛湿蠲痹法治疗寒湿型类风湿关节炎的临床研究[J].中国中医骨伤科杂志,2006,14(1):40-42.
[11]庞学丰,刘欢,吴燕红,等.补肾抗风湿中药联合西药治疗类风湿关节炎继发骨质疏松临床研究[J].世界中西医结合杂志,2015,10(1):47-49.
[12]庞学丰,任志宏,程世和,等.寒痹康汤对实验性关节炎大鼠血清IL-1β及IL-4的影响[J].陕西中医,2009,30(7):919-920.
[13]庞学丰,王乾,吴燕红,等.寒痹康汤对胶原诱导性关节炎大鼠滑膜细胞NF-κB表达的影响[J].风湿病与关节炎,2013,2(7):32-34.
[14]庞学丰,舒建龙,李玉玲,等.寒痹康汤对胶原诱导性关节炎大鼠血清VEGF的影响[J].陕西中医,2015,36(5):616-618.