膝骨性关节炎模型大鼠接受推拿治疗后软骨Toll样受体4/髓样分化因子88信号转导通路的变化
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摘要
背景:研究表明,Toll样受体4/髓样分化因子88信号转导通路可诱导各类炎症反应,且膝骨关节炎的发生与Toll样受体4/髓样分化因子88信号转导通路密切相关,而推拿可以降低白细胞介素6及肿瘤坏死因子α等炎症因子的表达水平,起到消炎止痛的作用,且对膝骨关节炎防治有一定的疗效。目的:对建立的大鼠膝骨关节炎模型行推拿治疗,观察其软骨Toll样受体4、髓样分化因子88、肿瘤坏死因子受体相关因子6、核转录因子κBp65 mRNA及蛋白表达,来探讨推拿治疗膝骨关节炎的效果及其机制。方法:采用随机化的原则,抽取10只大鼠为空白组,余下40只大鼠采用木瓜蛋白酶关节注射建立大鼠膝骨关节炎模型,待造模成功后,再将造模大鼠随机分为模型组、推拿组、塞来昔布组、推拿加塞来昔布组,每组10只。空白组、模型组常规饲养,推拿组一指禅手法治疗,塞来昔布组塞来昔布灌胃,推拿加塞来昔布组予以上2种治疗,各组均治疗4周。结果与结论:①与模型组相比,推拿和/或塞来昔布治疗均能显著降低大鼠软骨Mankin评分;②与模型组相比,推拿和/或塞来昔布治疗均能明显下调大鼠软骨组织中Toll样受体4、髓样分化因子88、核转录因子κΒp65以及肿瘤坏死因子受体相关因子6 mRNA与蛋白的表达水平,且推拿加塞来昔布组软骨组织中Toll样受体4、髓样分化因子88以及核转录因子κΒp65 mRNA与空白组接近;③提示推拿防治膝骨关节炎的机制可能是通过下调Toll样受体4、髓样分化因子88、肿瘤坏死因子受体相关因子6、核转录因子κΒp65 mRNA和蛋白的表达,从而延缓膝骨关节炎的发展进程。
BACKGROUND: Toll-like receptor 4/myeloid differentiation factor 88 signal transduction pathway has been shown to induce various inflammatory reactions, and the occurrence of knee osteoarthritis is closely related to Toll-like receptor 4/myeloid differentiation factor 88 signal transduction pathway. Massage can reduce inflammatory factors such as interleukin 6 and tumor necrosis factor alpha to exert anti-inflammatory and analgesic role, and has a certain effect on the prevention and treatment of knee osteoarthritis. OBJECTIVE: To explore the expression of Toll-like receptor 4, myeloid differentiation factor 88, tumor necrosis factor receptor associated factor 6 and nuclear factor kappa B p65 mRNA and protein in a rat model of knee osteoarthritis treated with massage, and to explore the effect and mechanism of massage in the treatment of knee osteoarthritis. METHODS: Ten rats were randomized into blank group and the remaining 40 rats were injected with papain to establish knee osteoarthritis model. The rat models were randomly divided into model, massage, celecoxib and massage plus celecoxib groups(n=10/group). The blank and model groups were fed routinely. The massage group was treated with massage using one finger. The celecoxib group was given intragastric administration of celecoxib. The massage plus celecoxib group was given the above two treatments. Each group was treated for 4 weeks. RESULTS AND CONCLUSION:(1) Compared with the model group, massage and/or celecoxib treatment could significantly reduce the Mankin score of cartilage in rats.(2) Compared with the model group, massage and/or celecoxib treatment could significantly reduce the expression levels of Toll-like receptor 4, myeloid differentiation factor 88, nuclear factor kappa B p65 and tumor necrosis factor receptor associated factor 6 mRNA and protein in cartilage tissue of rats. The expression levels of above cytokines in the massage and celecoxib group were similar to the blank group.(3) These results suggest that massage prevents and treats knee osteoarthritis probably through down-regulating the expression of Toll-like receptor 4, myeloid differentiation factor 88, tumor necrosis factor receptor associated factor 6, nuclear factor kappa B p65 mRNA and protein, thus delaying the progression of knee osteoarthritis.
BACKGROUND: Toll-like receptor 4/myeloid differentiation factor 88 signal transduction pathway has been shown to induce various inflammatory reactions, and the occurrence of knee osteoarthritis is closely related to Toll-like receptor 4/myeloid differentiation factor 88 signal transduction pathway. Massage can reduce inflammatory factors such as interleukin 6 and tumor necrosis factor alpha to exert anti-inflammatory and analgesic role, and has a certain effect on the prevention and treatment of knee osteoarthritis. OBJECTIVE: To explore the expression of Toll-like receptor 4, myeloid differentiation factor 88, tumor necrosis factor receptor associated factor 6 and nuclear factor kappa B p65 mRNA and protein in a rat model of knee osteoarthritis treated with massage, and to explore the effect and mechanism of massage in the treatment of knee osteoarthritis. METHODS: Ten rats were randomized into blank group and the remaining 40 rats were injected with papain to establish knee osteoarthritis model. The rat models were randomly divided into model, massage, celecoxib and massage plus celecoxib groups(n=10/group). The blank and model groups were fed routinely. The massage group was treated with massage using one finger. The celecoxib group was given intragastric administration of celecoxib. The massage plus celecoxib group was given the above two treatments. Each group was treated for 4 weeks. RESULTS AND CONCLUSION:(1) Compared with the model group, massage and/or celecoxib treatment could significantly reduce the Mankin score of cartilage in rats.(2) Compared with the model group, massage and/or celecoxib treatment could significantly reduce the expression levels of Toll-like receptor 4, myeloid differentiation factor 88, nuclear factor kappa B p65 and tumor necrosis factor receptor associated factor 6 mRNA and protein in cartilage tissue of rats. The expression levels of above cytokines in the massage and celecoxib group were similar to the blank group.(3) These results suggest that massage prevents and treats knee osteoarthritis probably through down-regulating the expression of Toll-like receptor 4, myeloid differentiation factor 88, tumor necrosis factor receptor associated factor 6, nuclear factor kappa B p65 mRNA and protein, thus delaying the progression of knee osteoarthritis.
引文
[1]胥少汀,葛宝丰,徐印钦.实用骨科学[M].4版.北京:人民军医出版社,2012:1676-1682.
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[3]李建华,龚利,房敏,等.推拿对膝骨关节炎患者膝屈伸肌肌张力的影响[J].中国骨伤,2011,24(7):575-577.
[4]杜宁,陆勇,顾翔,等.手法促进膝关节炎软骨修复的核磁共振病例对照研究[J].中国骨伤,2008,21(11):824-827.
[5]方苏亭,王胜利.松凝分筋手法治疗兔膝骨性关节炎病的实验研究[J].中国中医骨伤科杂志,2001,9(2):27.
[6]Pat Archer.运动保健按摩疗法[M].李国惠,译.天津:天津科技翻译出版公司,2008:70.
[7]王春林,向勇,田启东,等.“拔伸松动手法”对膝骨性关节炎兔白细胞介素1β和肿瘤坏死因子α的影响[J].环球中医药,2014,7(2):85-88.
[8]韦嵩,申茜茸,李晓昊,等.经筋微创松解术治疗膝骨关节炎对TLR4/MyD88/NF-κB信号转导通路的影响[J].中华中医药杂志, 2018,33(10):4637-4641.
[9]郑洁,赵莉平,胡亚莉,等.青藤碱对兔膝骨关节炎模型软骨Toll样受体2、4及髓样分化因子88表达的影响[J].中国中医药信息杂志,2018,25(9):49-51.
[10]李忠仁.实验针灸学[M].北京:中国中医药出版社,2008:33-35.
[11]魏伟,吴希美,李元建.药理实验方法学[M].4版.北京:人民卫生出版社,2010:1698.
[12]王文胜,陈伟,沈堡垒,等.上、下坡跑台训练膝骨关节炎模型大鼠关节软骨及炎症因子的变化[J].中国组织工程研究, 2016,20(2):197-200.
[13]Mankin HJ. Biochemical and metabolic abnormalities in osteoarthritic human cartilage. Fed Proc. 1973;32(4):1478-1480.
[14]杨智,程春凤,黄昕艳,等.Trizol法提取新生小鼠脑组织总RNA优化比较[J].中风与神经疾病杂志,2012,29(2):155-156.
[15]Sofat N, Robertson SD, Wait R. Fibronectin III 13-14 domains induce joint damage via Toll-like receptor 4 activation and synergize with interleukin-1and tumour necrosis factor. J Innate Immun. 2012;4(1):69-79.
[16]胡炳麟,龚利,李建华,等.一指禅推法治疗膝关节骨性关节炎的临床研究[J].中医药导报,2017,23(2):72-74.
[17]朱雪坤,杨日芳,孟艳秋.Toll样受体4配体的研究进展[J].中国药理学与毒理学杂志,2016,30(4):389-396.
[18]Temperley ND, Berlin S, Paton IR, et al. Evolution of the chicken Toll-like receptor gene family:a story of gene gain and gene loss. BMC Genomics. 2008;9:62.
[19]Anwar MA, Basith S, Choi S. Negative regulatory approaches to the attenuation of Toll-like receptor signaling. Exp Mol Med. 2013;45:e11.
[20]钱臣,卞晓星.Toll样受体4表达与膝骨关节炎软骨细胞凋亡的关系[J].江苏大学学报,2015,25(2):128-132.
[21]Hua F, Ha T, Ma J, et al. Blocking the MyD88-dependent pathway protects the myocardium from ischemia/reperfusion injury in rat hearts.Biochem Biophys Res Commun. 2005;338(2):1118-1125.
[22]Spehlmann ME, Eckmann L. Nuclear factor-kappa B in intestinal protection and destruction. Curr Opin Gastroenterol. 2009;25(2):92-99.
[23]Lim BJ, Lee D, Hong SW, et al. Toll-like receptor 4 signaling is involved in IgA-stimulated mesangial cell activation. Yonsei Med J. 2011;52(4):610-615.
[24]Ye H, Arron JR, Lamothe B, et al. Distinct molecular mechanism for initiating TRAF6 signalling. Nature. 2002;418(6896):443-447.
[25]Walsh MC, Lee J, Choi Y. Tumor necrosis factor receptorassociated factor 6(TRAF6)regulation of development, function,and homeostasis of the immune system. Immunol Rev. 2015;266(1):72-92.
[26]Sina C, Arlt A, Gavrilova O, et al. Ablation of gly96/immediate early gene-X1(gly96/iex-1)aggravates DSS-induced colitis in mice:role for gly96/iex-1 in the regulation of NF-kappaB. Inflamm Bowel Dis. 2010;16(2):320-331.
[27]Ostuni R, Zanoni I, Granucci F. Deciphering the complexity of Toll-like receptor signaling. Cell Mol Life Sci. 2010;67(24):4109-4134.
[28]Yang J, Amiri KI, Burke JR, et al. BMS-345541 targets inhibitor of kappaB kinase and induces apoptosis in melanoma:involvement of nuclear factor kappaB and mitochondria pathways. Clin Cancer Res.2006;12(3 Pt 1):950-960.
[29]何璐,吴子建,胡玲,等.艾灸对膝骨性关节炎模型大鼠膝关节软组织中NF-kbp65蛋白和IKB-α蛋白的影响[J].中医药临床杂志,2017,29(6):856-858.
[30]袁普卫,李小群,康武林,等.蠲痹胶囊对膝骨关节炎豚鼠关节软骨组织形态及滑膜中Toll样受体4?NF-κB p65及肿瘤坏死因子-α表达的影响[J].中医正骨,2016,28(9):5-12.
[2]严隽陶.推拿学[M].北京:中国中医药出版社,2003:175-176.
[3]李建华,龚利,房敏,等.推拿对膝骨关节炎患者膝屈伸肌肌张力的影响[J].中国骨伤,2011,24(7):575-577.
[4]杜宁,陆勇,顾翔,等.手法促进膝关节炎软骨修复的核磁共振病例对照研究[J].中国骨伤,2008,21(11):824-827.
[5]方苏亭,王胜利.松凝分筋手法治疗兔膝骨性关节炎病的实验研究[J].中国中医骨伤科杂志,2001,9(2):27.
[6]Pat Archer.运动保健按摩疗法[M].李国惠,译.天津:天津科技翻译出版公司,2008:70.
[7]王春林,向勇,田启东,等.“拔伸松动手法”对膝骨性关节炎兔白细胞介素1β和肿瘤坏死因子α的影响[J].环球中医药,2014,7(2):85-88.
[8]韦嵩,申茜茸,李晓昊,等.经筋微创松解术治疗膝骨关节炎对TLR4/MyD88/NF-κB信号转导通路的影响[J].中华中医药杂志, 2018,33(10):4637-4641.
[9]郑洁,赵莉平,胡亚莉,等.青藤碱对兔膝骨关节炎模型软骨Toll样受体2、4及髓样分化因子88表达的影响[J].中国中医药信息杂志,2018,25(9):49-51.
[10]李忠仁.实验针灸学[M].北京:中国中医药出版社,2008:33-35.
[11]魏伟,吴希美,李元建.药理实验方法学[M].4版.北京:人民卫生出版社,2010:1698.
[12]王文胜,陈伟,沈堡垒,等.上、下坡跑台训练膝骨关节炎模型大鼠关节软骨及炎症因子的变化[J].中国组织工程研究, 2016,20(2):197-200.
[13]Mankin HJ. Biochemical and metabolic abnormalities in osteoarthritic human cartilage. Fed Proc. 1973;32(4):1478-1480.
[14]杨智,程春凤,黄昕艳,等.Trizol法提取新生小鼠脑组织总RNA优化比较[J].中风与神经疾病杂志,2012,29(2):155-156.
[15]Sofat N, Robertson SD, Wait R. Fibronectin III 13-14 domains induce joint damage via Toll-like receptor 4 activation and synergize with interleukin-1and tumour necrosis factor. J Innate Immun. 2012;4(1):69-79.
[16]胡炳麟,龚利,李建华,等.一指禅推法治疗膝关节骨性关节炎的临床研究[J].中医药导报,2017,23(2):72-74.
[17]朱雪坤,杨日芳,孟艳秋.Toll样受体4配体的研究进展[J].中国药理学与毒理学杂志,2016,30(4):389-396.
[18]Temperley ND, Berlin S, Paton IR, et al. Evolution of the chicken Toll-like receptor gene family:a story of gene gain and gene loss. BMC Genomics. 2008;9:62.
[19]Anwar MA, Basith S, Choi S. Negative regulatory approaches to the attenuation of Toll-like receptor signaling. Exp Mol Med. 2013;45:e11.
[20]钱臣,卞晓星.Toll样受体4表达与膝骨关节炎软骨细胞凋亡的关系[J].江苏大学学报,2015,25(2):128-132.
[21]Hua F, Ha T, Ma J, et al. Blocking the MyD88-dependent pathway protects the myocardium from ischemia/reperfusion injury in rat hearts.Biochem Biophys Res Commun. 2005;338(2):1118-1125.
[22]Spehlmann ME, Eckmann L. Nuclear factor-kappa B in intestinal protection and destruction. Curr Opin Gastroenterol. 2009;25(2):92-99.
[23]Lim BJ, Lee D, Hong SW, et al. Toll-like receptor 4 signaling is involved in IgA-stimulated mesangial cell activation. Yonsei Med J. 2011;52(4):610-615.
[24]Ye H, Arron JR, Lamothe B, et al. Distinct molecular mechanism for initiating TRAF6 signalling. Nature. 2002;418(6896):443-447.
[25]Walsh MC, Lee J, Choi Y. Tumor necrosis factor receptorassociated factor 6(TRAF6)regulation of development, function,and homeostasis of the immune system. Immunol Rev. 2015;266(1):72-92.
[26]Sina C, Arlt A, Gavrilova O, et al. Ablation of gly96/immediate early gene-X1(gly96/iex-1)aggravates DSS-induced colitis in mice:role for gly96/iex-1 in the regulation of NF-kappaB. Inflamm Bowel Dis. 2010;16(2):320-331.
[27]Ostuni R, Zanoni I, Granucci F. Deciphering the complexity of Toll-like receptor signaling. Cell Mol Life Sci. 2010;67(24):4109-4134.
[28]Yang J, Amiri KI, Burke JR, et al. BMS-345541 targets inhibitor of kappaB kinase and induces apoptosis in melanoma:involvement of nuclear factor kappaB and mitochondria pathways. Clin Cancer Res.2006;12(3 Pt 1):950-960.
[29]何璐,吴子建,胡玲,等.艾灸对膝骨性关节炎模型大鼠膝关节软组织中NF-kbp65蛋白和IKB-α蛋白的影响[J].中医药临床杂志,2017,29(6):856-858.
[30]袁普卫,李小群,康武林,等.蠲痹胶囊对膝骨关节炎豚鼠关节软骨组织形态及滑膜中Toll样受体4?NF-κB p65及肿瘤坏死因子-α表达的影响[J].中医正骨,2016,28(9):5-12.
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