针刺对实验性自身免疫性脑脊髓炎小鼠JAK2、P-STAT3、IL-17表达的影响
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摘要
目的:观察针刺干预C57BL/6小鼠实验性自身免疫性脑脊髓炎(EAE)的疗效及其对JAK2、PSTAT3和IL-17表达的影响,为临床治疗MS提供新的思路及理论支持。方法:采用髓鞘少突胶质细胞糖蛋白(MOG35-55)制备的完全抗原将C57BL/6小鼠诱导成EAE模型,将40只C57BL/6小鼠随机分为空白对照组、EAE模型组、激素治疗组、针刺治疗组,记录各组0天、14天、20天及28天的体质量、神经功能缺损评分观察临床症状,免疫第28天取材后脑组织进行LFB髓鞘染色观察病理变化,检测脑组织JAK2、P-STAT3免疫组化阳性表达及血清中IL-17ELISA表达。结果:与空白对照组相比,各组的体质量明显降低(P <0. 01),神经功能缺损评分明显升高(P <0. 01),脑组织LFB染色脱髓鞘程度明显升高,脑组织免疫组化JAK2、P-STAT3表达显著升高(P <0. 01),血清IL-17表达显著升高(P <0. 01)。与EAE模型组相比,针刺治疗组、激素治疗组的体质量明显增高(P <0. 01),神经功能缺损评分明显降低(P <0. 05或P <0. 01),脑组织LFB染色脱髓鞘程度明显降低,JAK2、P-STAT3表达显著降低(P <0. 01),IL-17表达显著降低(P <0. 05或P <0. 01)。与激素治疗组相比,针刺治疗组体质量较低(P <0. 05),神经功能缺损评分较高(免疫20天,P <0. 01;免疫28天,P> 0. 05),JAK2、PSTAT3表达较高但差异无统计学意义(P> 0. 05),IL-17表达较高(P <0. 05)。结论:针刺可能是通过降低EAE小鼠JAK2、P-STAT3、IL-17表达,调节特异性免疫应答,减轻炎症反应,改善髓鞘脱失程度,从而发挥治疗作用。
Objective: To observe the effect of acupuncture on experimental autoimmune encephalomyelitis( EAE) in C57 BL/6 mice and its effect on the expressions of JAK2,P-STAT3 and IL-17,thus to provide new ideas and theoretical support for clinical treatment of multiple sclerosis( MS). Methods: EAE model was established in C57 BL/6 mice by complete antigen prepared from myelin oligodendrocyte glycoprotein( MOG35-55). 40 mice of C57 BL/6 were randomly divided into the blank control group,the EAE model group,the hormone treatment group and the acupuncture treatment group. The weight and neurological function score of each group were recorded to observe the clinical symptoms. On the 28 th day,the pathological changes in brain tissue were observed by Luxol Fast Blue( LFB) myelin staining method,the expressions of JAK2 and P-STAT3 in brain tissue were measured by immunohistochemistry method,and the expression of IL-17 in serum was detected by ELISA method. Results: Compared to the blank control group,the weights of the other groups were significantly lower( P < 0. 01); the neurological function scores were significantly higher( P < 0. 01); the degrees of demyelination were significantly increased; the expressions of JAK2 and P-STAT3 were significantly increased( P < 0. 01); the serum expressions of IL-17 were significantly increased( P < 0. 01). Compared to the EAE model group,the weights of the acupuncture treatment group and the hormone treatment group were significantly increased( P < 0. 01); the neurological function scores were significantly decreased( P < 0. 05,P <0. 01) and the degrees of demyelination were decreased; the expressions of JAK2 and P-STAT3 were significantly decreased( P < 0. 01),and the serum expressions of IL-17 were significantly decreased( P < 0. 05,P <0. 01). Compared to the hormone treatment group,the weight of the acupuncture treatment group was lower( P < 0. 05); the neurological function score was a bit higher( 20 d,P < 0. 01; 28 d,P < 0. 05); there were no statistical differences in the expressions of JAK2 and P-STAT3 between the two groups( P > 0. 05); the serum expression of IL-17 was higher( P < 0. 05). Conclusion: Acupuncture has a therapeutic effect on EAE mice,which can reduce the specific immune response,the inflammatory response,as well as reduce the degree of demyelinstion and the expressions of JAK2,P-STAT3 and IL-17.
Objective: To observe the effect of acupuncture on experimental autoimmune encephalomyelitis( EAE) in C57 BL/6 mice and its effect on the expressions of JAK2,P-STAT3 and IL-17,thus to provide new ideas and theoretical support for clinical treatment of multiple sclerosis( MS). Methods: EAE model was established in C57 BL/6 mice by complete antigen prepared from myelin oligodendrocyte glycoprotein( MOG35-55). 40 mice of C57 BL/6 were randomly divided into the blank control group,the EAE model group,the hormone treatment group and the acupuncture treatment group. The weight and neurological function score of each group were recorded to observe the clinical symptoms. On the 28 th day,the pathological changes in brain tissue were observed by Luxol Fast Blue( LFB) myelin staining method,the expressions of JAK2 and P-STAT3 in brain tissue were measured by immunohistochemistry method,and the expression of IL-17 in serum was detected by ELISA method. Results: Compared to the blank control group,the weights of the other groups were significantly lower( P < 0. 01); the neurological function scores were significantly higher( P < 0. 01); the degrees of demyelination were significantly increased; the expressions of JAK2 and P-STAT3 were significantly increased( P < 0. 01); the serum expressions of IL-17 were significantly increased( P < 0. 01). Compared to the EAE model group,the weights of the acupuncture treatment group and the hormone treatment group were significantly increased( P < 0. 01); the neurological function scores were significantly decreased( P < 0. 05,P <0. 01) and the degrees of demyelination were decreased; the expressions of JAK2 and P-STAT3 were significantly decreased( P < 0. 01),and the serum expressions of IL-17 were significantly decreased( P < 0. 05,P <0. 01). Compared to the hormone treatment group,the weight of the acupuncture treatment group was lower( P < 0. 05); the neurological function score was a bit higher( 20 d,P < 0. 01; 28 d,P < 0. 05); there were no statistical differences in the expressions of JAK2 and P-STAT3 between the two groups( P > 0. 05); the serum expression of IL-17 was higher( P < 0. 05). Conclusion: Acupuncture has a therapeutic effect on EAE mice,which can reduce the specific immune response,the inflammatory response,as well as reduce the degree of demyelinstion and the expressions of JAK2,P-STAT3 and IL-17.
引文
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[17]Kim KY,Yang WJ,Shin TK,et al.Impact of acupuncture by using lifeenergy(qi)oriental needle on the paralysis of rats with experimental autoimmune encephalomyelitis[J].Am J Chin Med,2012,40(4):769.
[18]Liu YM,Liu XJ,Bai SS,et al.The effect of electroacupuncture on Tcell responses in rats with experimental autoimmune encephalitis[J].JNeuroimmunol,2010,220(1-2):25.
[19]Lee MJ,Jang M,Choi J,et al.Bee Venom Acupuncture Alleviates Experimental Autoimmune Encephalomyelitis by Upregulating Regulatory T Cells and Suppressing Th1 and Th17 Responses[J].Mol Neurobiol,2016,53(3):1419.
[20]关婧垚.头针结合运动训练对EAE小鼠脾淋巴细胞Th17和Treg的影响[D].哈尔滨:黑龙江中医药大学,2013.
[21]Liu Y,Wang H,Wang X,et al.The mechanism of effective electroacupuncture on T cell response in rats with experimental autoimmune encephalomyelitis[J].PLo S One,2013,8(1):e51573.
[22]董英.头针结合运动训练对EAE小鼠血清中IL-17和TGF-β1的影响[D].哈尔滨:黑龙江中医药大学,2013.
[23]Liu R,Xu N,Yi W,et al.Electroacupuncture effect on neurological behavior and tyrosine kinase-JAK 2 in rats with focal cerebral ischemia[J].J Tradit Chin Med,2012,32(3):465.
[24]杨丽萍,王明臣,刘旺根,等.温针灸对虚寒型膝骨关节炎基因表达通路的影响[J].中国针灸,2007,27(9):677-680.
[25]刘智斌,杨晓航.不同手法针刺足三里穴对人PBMC的STAT5信号转导途径的作用[J].中国针灸,2006,26(2):46-48.
[2]Krementsov DN,Thornton TM,Teuscher C,et al.The emerging role of p38 mitogen-activated protein kinase in multiple sclerosis and its models[J].Mol Cell Biol,2013,33(19):3728.
[3]Benveniste EN,Liu Y,Mcfarland BC,et al.Involvement of the janus kinase/signal transducer and activator of transcription signaling pathway in multiple sclerosis and the animal model of experimental autoimmune encephalomyelitis[J].J Interferon Cytokine Res,2014,34(8):577.
[4]Yin L,Chen Y,Qu Z,et al.Involvement of JAK/STAT signaling in the effect of cornel iridoid glycoside on experimental autoimmune encephalomyelitis amelioration in rats[J].J Neuroimmunol,2014,274(1-2):28-37.
[5]中华医学会神经病学分会神经免疫学组.多发性硬化诊断和治疗中国专家共识(2014版)[J].中华神经科杂志,2015,48(5):362-367.
[6]Stoll SS,Nieves C,Tabby DS,et al.Use of therapies other than diseasemodifying agents,including complementary and alternative medicine,by patients with multiple sclerosis:a survey study[J].J Am Osteopath Assoc,2012,112(1):22.
[7]Esmonde L,Long AF.Complementary therapy use by persons with multiple sclerosis:benefits and research priorities[J].Complement Ther Clin Pract,2008,14(3):176.
[8]Karpatkin HI,Napolione D,Siminovichblok B.Acupuncture and Multiple Sclerosis:A Review of the Evidence[J].Evidence-Based Complementray and Alternative Medicine,2014(2):972935.
[9]冯美娜,陈玉华.多发性硬化的联合治疗进展[J].医学综述,2015,22(18):3361-3363.
[10]Denic A,Johnson AJ,Bieber AJ,et al.The relevance of animal models in multiple sclerosis research[J].Pathophysiology,2011,18(1):21.
[11]Jia Y,Jing J,Bai Y,et al.Amelioration of experimental autoimmune encephalomyelitis by plumbagin through down-regulation of JAK-STAT and NF-kappa B signaling pathways[J].PLo S One,2011,6(10):e27006.
[12]Jadidi NF,Mirshafiey A.Th17 cell,the new player of neuroinflammatory process in multiple sclerosis[J].Scand J Immunol,2011,74(1):1.
[13]Villarino AV,Kanno Y,Ferdinand JR,et al.Mechanisms of Jak/STATsignaling in immunity and disease[J].J Immunol,2015,194(1):21.
[14]Shaposhnikov AV,Komar'KIF,Lebedeva LA,et al.Molecular components of JAK/STAT signaling pathway and its connection with transcription machinery[J].Mol Biol(Mosk),2013,47(3):388.
[15]周莉,樊永平,叶明.59例多发性硬化患者不同中医证型的免疫学研究[J].中国中西医结合杂志,2007,27(7):599-601.
[16]关婧垚,唐强,李德生,等.多发性硬化的针灸、康复治疗及瞻望[J].科学技术创新,2013(4):36-37.
[17]Kim KY,Yang WJ,Shin TK,et al.Impact of acupuncture by using lifeenergy(qi)oriental needle on the paralysis of rats with experimental autoimmune encephalomyelitis[J].Am J Chin Med,2012,40(4):769.
[18]Liu YM,Liu XJ,Bai SS,et al.The effect of electroacupuncture on Tcell responses in rats with experimental autoimmune encephalitis[J].JNeuroimmunol,2010,220(1-2):25.
[19]Lee MJ,Jang M,Choi J,et al.Bee Venom Acupuncture Alleviates Experimental Autoimmune Encephalomyelitis by Upregulating Regulatory T Cells and Suppressing Th1 and Th17 Responses[J].Mol Neurobiol,2016,53(3):1419.
[20]关婧垚.头针结合运动训练对EAE小鼠脾淋巴细胞Th17和Treg的影响[D].哈尔滨:黑龙江中医药大学,2013.
[21]Liu Y,Wang H,Wang X,et al.The mechanism of effective electroacupuncture on T cell response in rats with experimental autoimmune encephalomyelitis[J].PLo S One,2013,8(1):e51573.
[22]董英.头针结合运动训练对EAE小鼠血清中IL-17和TGF-β1的影响[D].哈尔滨:黑龙江中医药大学,2013.
[23]Liu R,Xu N,Yi W,et al.Electroacupuncture effect on neurological behavior and tyrosine kinase-JAK 2 in rats with focal cerebral ischemia[J].J Tradit Chin Med,2012,32(3):465.
[24]杨丽萍,王明臣,刘旺根,等.温针灸对虚寒型膝骨关节炎基因表达通路的影响[J].中国针灸,2007,27(9):677-680.
[25]刘智斌,杨晓航.不同手法针刺足三里穴对人PBMC的STAT5信号转导途径的作用[J].中国针灸,2006,26(2):46-48.