牛膝总皂苷干预兔膝骨关节炎滑膜液来源细胞因子的表达
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摘要
背景:既往研究表明,滑膜液中细胞因子是引起膝关节疼痛的重要因素。牛膝能够促进膝关节软骨细胞修复,降低膝关节炎症水平,减轻膝关节疼痛。牛膝能否调控膝关节炎滑膜液内细胞因子水平,目前尚不清楚。目的:探讨牛膝总皂苷对实验兔膝骨关节炎滑膜液来源细胞因子谱的影响。方法:8只新西兰兔随机分为实验组和对照组,每组4只,均采用石膏外固定的方法构建右膝关节炎模型,造模后第2天开始给予牛膝总皂苷及蒸馏水灌服,灌服2周后测定关节活动度,抽取膝关节滑膜液,采用Luminex法测定炎性细胞因子和趋化因子水平,采用ELISA法检测基质金属蛋白酶1、基质金属蛋白酶2、基质金属蛋白酶3水平。实验经广州中医药大学实验动物伦理委员会批准(批准号:2018-31)。结果与结论:(1)治疗后,实验组膝关节活动度明显大于对照组(P <0.05);(2)与对照组相比,实验组治疗后炎性因子白细胞介素1α、白细胞介素1β、白细胞介素10、肿瘤坏死因子α,趋化因子CCL3、CXCL8、CXCL9,基质金属蛋白酶1、基质金属蛋白酶2、基质金属蛋白酶3的表达水平均降低,差异有显著性意义(P <0.05);(3)结果表明,牛膝总皂苷能够缓解骨关节炎兔膝关节活动障碍,降低膝关节滑膜液中细胞因子水平,从而起到减轻炎症和保护软骨的作用。
BACKGROUND: Previous studies have shown that synovial fluid cytokines are important factors responsible for knee pain. Achyranthes bidentata can promote the repair of knee chondrocytes, relieve inflammation of the knee joint, and ease knee pain. Whether Achyranthes bidentata can regulate the level of cytokines in the synovial fluid of knee osteoarthritis is still unclear. OBJECTIVE: To investigate the effect of total saponins of Achyranthes bidentata on cytokine profiles of synovial fluid in experimental rabbits with knee osteoarthritis.METHODS: Eight New Zealand rabbits were randomized into experimental and control groups(n=4/group). A rabbit model of knee osteoarthritis was established. At 2 days after modeling, the total saponins of Achyranthes bidentata and distilled water were administered in the experimental and control group, respectively. Range of motion of the knee joint was measured after 2 weeks of administration. The synovial fluid of the knee joint was extracted from each rabbit. The levels of inflammatory cytokines and chemokines were measured by Luminex. The levels of matrix metalloproteinases 1, 2, 3 mRNA were measured by ELISA. The study protocol was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine(approval No. 2018-31). RESULTS AND CONCLUSION: The range of motion of the knee joint in the experimental group was significantly higher than that in the control group(P < 0.05). Compared with the control group, the expression levels of inflammatory factors interleukin 1α, interleukin 1β, interleukin 10, tumor necrosis factor α, chemokines CCL3, CXCL8, CXCL9, matrix metalloproteinases 1, 2, 3 decreased significantly after treatment(P < 0.05). These findings indicate that Achyranthes bidentata total saponins can alleviate knee dysfunction in knee osteoarthritis rabbits, and reduce the level of cytokines in the synovial fluid of the knee joint, thus reducing inflammation and protecting the cartilage.
BACKGROUND: Previous studies have shown that synovial fluid cytokines are important factors responsible for knee pain. Achyranthes bidentata can promote the repair of knee chondrocytes, relieve inflammation of the knee joint, and ease knee pain. Whether Achyranthes bidentata can regulate the level of cytokines in the synovial fluid of knee osteoarthritis is still unclear. OBJECTIVE: To investigate the effect of total saponins of Achyranthes bidentata on cytokine profiles of synovial fluid in experimental rabbits with knee osteoarthritis.METHODS: Eight New Zealand rabbits were randomized into experimental and control groups(n=4/group). A rabbit model of knee osteoarthritis was established. At 2 days after modeling, the total saponins of Achyranthes bidentata and distilled water were administered in the experimental and control group, respectively. Range of motion of the knee joint was measured after 2 weeks of administration. The synovial fluid of the knee joint was extracted from each rabbit. The levels of inflammatory cytokines and chemokines were measured by Luminex. The levels of matrix metalloproteinases 1, 2, 3 mRNA were measured by ELISA. The study protocol was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine(approval No. 2018-31). RESULTS AND CONCLUSION: The range of motion of the knee joint in the experimental group was significantly higher than that in the control group(P < 0.05). Compared with the control group, the expression levels of inflammatory factors interleukin 1α, interleukin 1β, interleukin 10, tumor necrosis factor α, chemokines CCL3, CXCL8, CXCL9, matrix metalloproteinases 1, 2, 3 decreased significantly after treatment(P < 0.05). These findings indicate that Achyranthes bidentata total saponins can alleviate knee dysfunction in knee osteoarthritis rabbits, and reduce the level of cytokines in the synovial fluid of the knee joint, thus reducing inflammation and protecting the cartilage.
引文
[1]Berenbaum F.Osteoarthritis as an inflammatory disease(osteoarthritis is not osteoarthrosis!).Osteoarthritis Cartilage.2013;21(1):16-21.
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[3]Greene MA,Loeser RF.Aging-related inflammation in osteoarthritis.Osteoarthritis Cartilage.2015;23(11):1966-1971.
[4]Mobasheri A,Bay-Jensen AC,van Spil WE,et al.Osteoarthritis Year in Review 2016:biomarkers(biochemical markers).Osteoarthritis Cartilage.2017;25(2):199-208.
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[8]陈达,廖州伟,马笃军,等.牛膝醇提物对兔骨关节炎模型的疗效比较[J].中国医药导报,2016,13(25):21-24.
[9]彭力平,马笃军,裴军宇.牛膝醇提物对急性软骨损伤兔软骨的影响[J].中医杂志,2013,54(17):1504-1506.
[10]曾俊华,马笃军,彭力平,等.实验兔膝骨关节炎模型的建立及鉴定[J].中国临床研究,2016,29(5):679-682.
[11]孙文才,白玉江,李玉成,等.骨关节炎患者血清及关节滑液中细胞因子的表达水平研究[J].中国全科医学,2012,15(27):3185-3186.
[12]史新立,胡堃,孟祥提.关节软骨修复与相关细胞因子的作用[J].中国组织工程研究与临床康复,2011,15(11):2047-2050.
[13]Vergunst CE,van de Sande MG,Lebre MC,et al.The role of chemokines in rheumatoid arthritis and osteoarthritis.Scand JRheumatol.2005;34(6):415-425.
[14]Wojdasiewicz P,Poniatowski?A,Szukiewicz D.The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of osteoarthritis.Mediators Inflamm.2014;2014:561459.
[15]Hedbom E,H?uselmann HJ.Molecular aspects of pathogenesis in osteoarthritis:the role of inflammation.Cell Mol Life Sci.2002;59(1):45-53.
[16]Schulze-Tanzil G,Zreiqat H,Sabat R,et al.Interleukin-10 and articular cartilage:experimental therapeutical approaches in cartilage disorders.Curr Gene Ther.2009;9(4):306-315.
[17]贺占坤,沈杰威.MMP-2、MMP-3、MMP-9和TIMP-1评价膝关节骨性关节炎的临床研究[J].重庆医学,2013,42(32):3872-3874.
[18]郭达,张斌山,刘金文.骨关节炎的古代方药规律研究[J].南方医科大学学报,2010,30(9):2194-2195.
[19]姚共和,刘向前,李建斌,等.中医期刊治疗膝关节骨关节炎方剂用药特点分析[J].湖南中医学院学报,2005,25(6):54-56.
[20]沈舒,王琼,李友宾.牛膝的化学成分和药理作用研究进展[J].海峡药学,2011,23(11):1-6.
[21]高晓燕,王大为,李发美,等.牛膝提取物对成骨样细胞增殖的作用[J].沈阳药科大学学报,2000,17(3):210-213.
[22]Lu P,Weaver VM,Werb Z.The extracellular matrix:a dynamic niche in cancer progression.J Cell Biol.2012;196(4):395-406.
[23]Konttinen YT,Ainola M,Valleala H,et al.Analysis of 16different matrix metalloproteinases(MMP-1 to MMP-20)in the synovial membrane:different profiles in trauma and rheumatoid arthritis.Ann Rheum Dis.1999;58(11):691-697.
[24]Adams SB,Setton LA,Bell RD,et al.Inflammatory Cytokines and Matrix Metalloproteinases in the Synovial Fluid After Intra-articular Ankle Fracture.Foot Ankle Int.2015;36(11):1264-1271.
[25]Jouglin M,Robert C,Valette JP,et al.Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses:correlation with articular cartilage alterations.Vet Res.2000;31(5):507-515.
[26]Ishizuka S,Sakai T,Hiraiwa H,et al.Hypoxia-inducible factor-2αinduces expression of type X collagen and matrix metalloproteinases 13 in osteoarthritic meniscal cells.Inflamm Res.2016;65(6):439-448.
[27]Yu WG,Shen Y,Wu JZ,et al.Madecassoside impedes invasion of rheumatoid fibroblast-like synoviocyte from adjuvant arthritis rats via inhibition of NF-κB-mediated matrix metalloproteinase-13 expression.Chin J Nat Med.2018;16(5):330-338.
[28]van Nieuwenhuijze AE,van de Loo FA,Walgreen B,et al.Complementary action of granulocyte macrophage colony-stimulating factor and interleukin-17A induces interleukin-23,receptor activator of nuclear factor-κB ligand,and matrix metalloproteinases and drives bone and cartilage pathology in experimental arthritis:rationale for combination therapy in rheumatoid arthritis.Arthritis Res Ther.2015;17:163.
[29]Taraboletti G,D'Ascenzo S,Borsotti P,et al.Shedding of the matrix metalloproteinases MMP-2,MMP-9,and MT1-MMP as membrane vesicle-associated components by endothelial cells.Am J Pathol.2002;160(2):673-680.
[30]Millward-Sadler SJ,Wright MO,Davies LW,et al.Mechanotransduction via integrins and interleukin-4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal,but not osteoarthritic,human articular chondrocytes.Arthritis Rheum.2000;43(9):2091-2099.
[31]Mengshol JA,Vincenti MP,Coon CI,et al.Interleukin-1induction of collagenase 3(matrix metalloproteinase 13)gene expression in chondrocytes requires p38,c-Jun N-terminal kinase,and nuclear factor kappaB:differential regulation of collagenase 1 and collagenase 3.Arthritis Rheum.2000;43(4):801-811.
[32]Vincenti MP,Brinckerhoff CE.Transcriptional regulation of collagenase(MMP-1,MMP-13)genes in arthritis:integration of complex signaling pathways for the recruitment of gene-specific transcription factors.Arthritis Res.2002;4(3):157-164.
[33]Hanemaaijer R,Sorsa T,Konttinen YT,et al.Matrix metalloproteinase-8 is expressed in rheumatoid synovial fibroblasts and endothelial cells.Regulation by tumor necrosis factor-alpha and doxycycline.J Biol Chem.1997;272(50):31504-31509.
[34]de Lange-Brokaar BJ,Ioan-Facsinay A,van Osch GJ,et al.Synovial inflammation,immune cells and their cytokines in osteoarthritis:a review.Osteoarthritis Cartilage.2012;20(12):1484-1499.
[2]Cross M,Smith E,Hoy D,et al.The global burden of hip and knee osteoarthritis:estimates from the global burden of disease 2010 study.Ann Rheum Dis.2014;73(7):1323-1330.
[3]Greene MA,Loeser RF.Aging-related inflammation in osteoarthritis.Osteoarthritis Cartilage.2015;23(11):1966-1971.
[4]Mobasheri A,Bay-Jensen AC,van Spil WE,et al.Osteoarthritis Year in Review 2016:biomarkers(biochemical markers).Osteoarthritis Cartilage.2017;25(2):199-208.
[5]Huang D,Liu YQ,Liang LS,et al.The Diagnosis and Therapy of Degenerative Knee Joint Disease:Expert Consensus from the Chinese Pain Medicine Panel.Pain Res Manag.2018;2018:2010129.
[6]Scanzello CR,Goldring SR.The role of synovitis in osteoarthritis pathogenesis.Bone.2012;51(2):249-257.
[7]肖伟,林栋栋,彭力平.牛膝醇提物透入疗法对膝骨性关节炎的疗效观察[J].中国中医骨伤科杂志,2015,23(8):37-40.
[8]陈达,廖州伟,马笃军,等.牛膝醇提物对兔骨关节炎模型的疗效比较[J].中国医药导报,2016,13(25):21-24.
[9]彭力平,马笃军,裴军宇.牛膝醇提物对急性软骨损伤兔软骨的影响[J].中医杂志,2013,54(17):1504-1506.
[10]曾俊华,马笃军,彭力平,等.实验兔膝骨关节炎模型的建立及鉴定[J].中国临床研究,2016,29(5):679-682.
[11]孙文才,白玉江,李玉成,等.骨关节炎患者血清及关节滑液中细胞因子的表达水平研究[J].中国全科医学,2012,15(27):3185-3186.
[12]史新立,胡堃,孟祥提.关节软骨修复与相关细胞因子的作用[J].中国组织工程研究与临床康复,2011,15(11):2047-2050.
[13]Vergunst CE,van de Sande MG,Lebre MC,et al.The role of chemokines in rheumatoid arthritis and osteoarthritis.Scand JRheumatol.2005;34(6):415-425.
[14]Wojdasiewicz P,Poniatowski?A,Szukiewicz D.The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of osteoarthritis.Mediators Inflamm.2014;2014:561459.
[15]Hedbom E,H?uselmann HJ.Molecular aspects of pathogenesis in osteoarthritis:the role of inflammation.Cell Mol Life Sci.2002;59(1):45-53.
[16]Schulze-Tanzil G,Zreiqat H,Sabat R,et al.Interleukin-10 and articular cartilage:experimental therapeutical approaches in cartilage disorders.Curr Gene Ther.2009;9(4):306-315.
[17]贺占坤,沈杰威.MMP-2、MMP-3、MMP-9和TIMP-1评价膝关节骨性关节炎的临床研究[J].重庆医学,2013,42(32):3872-3874.
[18]郭达,张斌山,刘金文.骨关节炎的古代方药规律研究[J].南方医科大学学报,2010,30(9):2194-2195.
[19]姚共和,刘向前,李建斌,等.中医期刊治疗膝关节骨关节炎方剂用药特点分析[J].湖南中医学院学报,2005,25(6):54-56.
[20]沈舒,王琼,李友宾.牛膝的化学成分和药理作用研究进展[J].海峡药学,2011,23(11):1-6.
[21]高晓燕,王大为,李发美,等.牛膝提取物对成骨样细胞增殖的作用[J].沈阳药科大学学报,2000,17(3):210-213.
[22]Lu P,Weaver VM,Werb Z.The extracellular matrix:a dynamic niche in cancer progression.J Cell Biol.2012;196(4):395-406.
[23]Konttinen YT,Ainola M,Valleala H,et al.Analysis of 16different matrix metalloproteinases(MMP-1 to MMP-20)in the synovial membrane:different profiles in trauma and rheumatoid arthritis.Ann Rheum Dis.1999;58(11):691-697.
[24]Adams SB,Setton LA,Bell RD,et al.Inflammatory Cytokines and Matrix Metalloproteinases in the Synovial Fluid After Intra-articular Ankle Fracture.Foot Ankle Int.2015;36(11):1264-1271.
[25]Jouglin M,Robert C,Valette JP,et al.Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses:correlation with articular cartilage alterations.Vet Res.2000;31(5):507-515.
[26]Ishizuka S,Sakai T,Hiraiwa H,et al.Hypoxia-inducible factor-2αinduces expression of type X collagen and matrix metalloproteinases 13 in osteoarthritic meniscal cells.Inflamm Res.2016;65(6):439-448.
[27]Yu WG,Shen Y,Wu JZ,et al.Madecassoside impedes invasion of rheumatoid fibroblast-like synoviocyte from adjuvant arthritis rats via inhibition of NF-κB-mediated matrix metalloproteinase-13 expression.Chin J Nat Med.2018;16(5):330-338.
[28]van Nieuwenhuijze AE,van de Loo FA,Walgreen B,et al.Complementary action of granulocyte macrophage colony-stimulating factor and interleukin-17A induces interleukin-23,receptor activator of nuclear factor-κB ligand,and matrix metalloproteinases and drives bone and cartilage pathology in experimental arthritis:rationale for combination therapy in rheumatoid arthritis.Arthritis Res Ther.2015;17:163.
[29]Taraboletti G,D'Ascenzo S,Borsotti P,et al.Shedding of the matrix metalloproteinases MMP-2,MMP-9,and MT1-MMP as membrane vesicle-associated components by endothelial cells.Am J Pathol.2002;160(2):673-680.
[30]Millward-Sadler SJ,Wright MO,Davies LW,et al.Mechanotransduction via integrins and interleukin-4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal,but not osteoarthritic,human articular chondrocytes.Arthritis Rheum.2000;43(9):2091-2099.
[31]Mengshol JA,Vincenti MP,Coon CI,et al.Interleukin-1induction of collagenase 3(matrix metalloproteinase 13)gene expression in chondrocytes requires p38,c-Jun N-terminal kinase,and nuclear factor kappaB:differential regulation of collagenase 1 and collagenase 3.Arthritis Rheum.2000;43(4):801-811.
[32]Vincenti MP,Brinckerhoff CE.Transcriptional regulation of collagenase(MMP-1,MMP-13)genes in arthritis:integration of complex signaling pathways for the recruitment of gene-specific transcription factors.Arthritis Res.2002;4(3):157-164.
[33]Hanemaaijer R,Sorsa T,Konttinen YT,et al.Matrix metalloproteinase-8 is expressed in rheumatoid synovial fibroblasts and endothelial cells.Regulation by tumor necrosis factor-alpha and doxycycline.J Biol Chem.1997;272(50):31504-31509.
[34]de Lange-Brokaar BJ,Ioan-Facsinay A,van Osch GJ,et al.Synovial inflammation,immune cells and their cytokines in osteoarthritis:a review.Osteoarthritis Cartilage.2012;20(12):1484-1499.