基于JAK2/STAT3信号通路探讨温针灸改善膝关节骨关节炎兔软骨损伤的机制  

Warm acupuncture stimulation improves cartilage damage and motor function by regulating JAK2/STAT3 signaling pathway in rabbits with knee osteoarthritis

作　　者：张艳玲[1] 刘君伟 李春[1] 马晓秀 武永利[1,2] ZHANG Yan-ling;LIU Jun-wei;LI Chun;MA Xiao-xiu;WU Yong-li(Department of Orthopedics and Traumatology of Traditional Chinese Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Ningxia Minority Medicine Modernization of Chinese Ministry of Education,Ningxia Medical University,Yinchuan 750004)

机构地区：[1]宁夏医科大学总医院中医骨伤科,银川750004 [2]宁夏医科大学宁夏少数民族医药现代化教育部重点实验室,银川750004

出　　处：《针刺研究》2022年第12期1088-1094,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.82160939);宁夏自然科学基金项目(No.2021AAC03347);宁夏回族自治区科技创新领军人才项目(No.KJT201610);宁夏医科大学校级项目(No.XZ2020021);第五批宁夏回族自治区青年科技人才托举工程。

摘　　要：目的:观察温针灸对膝关节骨关节炎(KOA)兔关节软骨Janus蛋白酪氨酸激酶2/信号转导和转录活化因子3(JAK2/STAT3)信号通路及炎性因子表达的影响,探讨温针灸治疗KOA的作用机制。方法:将新西兰兔随机分为空白组、模型组、温针灸组、药物组,每组12只。采用右后肢管型石膏伸直位固定法制备KOA兔模型。温针灸组选取“内膝眼”“外膝眼”“鹤顶”“足三里”予温针灸治疗15 min,药物组给予双氯芬酸钠溶液(0.35 mg/kg)灌胃,均每日1次,治疗4周。采用Lequesne MG量表对各组兔进行行为学评价;HE染色法观察膝关节软骨组织病理改变并进行Mankin评分;ELISA法检测血清白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶-9(MMP-9)的含量;Western blot法检测膝关节软骨组织p-JAK2/JAK2、p-STAT3/STAT3和MMP-9蛋白表达水平。结果:与空白组比较,模型组HE染色示膝关节软骨膜破损,表层细胞排列紊乱,Lequesne MG、Mankin评分及血清IL-6、TNF-α、MMP-9含量和膝关节软骨组织p-JAK2/JAK2、p-STAT3/STAT3比值和MMP-9蛋白表达水平均升高(P<0.01)。与模型组比较,温针灸组和药物组HE染色示膝关节软骨表面分布层次较清晰,Lequesne MG、Mankin评分及血清IL-6、TNF-α、MMP-9含量和膝关节软骨组织p-JAK2/JAK2、p-STAT3/STAT3比值和MMP-9蛋白表达水平均降低(P<0.01,P<0.05)。与药物组比较,温针灸组HE染色示膝关节软骨表面分布层次清晰,潮线结构完整,Lequesne MG、Mankin评分及血清IL-6、TNF-α、MMP-9含量和膝关节软骨组织p-JAK2/JAK2、p-STAT3/STAT3比值降低(P<0.01,P<0.05)。结论:温针灸可能通过抑制JAK2、STAT3及MMP-9蛋白表达,调控JAK2/STAT3信号通路,降低促炎因子分泌,减轻软骨损伤,发挥治疗作用。Objective To observe the effect of warm acupuncture on the expression of Janus protein tyrosine kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway and inflammatory factors of articular cartilage in rabbits with knee osteoarthritis(KOA),so as to explore its underlying mechanisms in improving KOA.Methods New Zea-land rabbits were randomly divided into blank,model,warm acupuncture and medication groups(12 rabbits in each group).The KOA model was prepared by using the right hind limb tubular plaster extension fixation method.The rabbits in the warm acupuncture group received acupuncture of“Neixiyan”(EX-LE4),“Waixiyan”(ST35),“Heding”(EX-LE2)and“Zusanli”(ST36),followed by attaching an ignited moxa-stick segment to the acupuncture-handle.The treatment was conducted for 15 min,once a week for 4 weeks.The rabbits in the medication group received gavage of diclofenac sodium solution(0.35 mg/kg),once daily for 4 weeks.The dysfunction severity state of the rabbit’s knee-joint was evaluated using Lequesne scale(0-3 points),and the histopathological changes of cartilage were observed under microscope after H.E.staining and the state of distribution of chondrocytes in different layers and the extracellular matrix was assessed using Mankin score(0-6 points).The contents of serum interleukin(IL)-6,tumor necrosis factor-α(TNF-α)and matrix metalloproteinase-9(MMP-9)were measured by using ELISA,and the expression levels of p-JAK2/JAK2,p-STAT3/STAT3 and MMP-9 in knee cartilage tissue were detected using Western blot.Results Compared with the blank group,the Lequesne score,Mankin score,and the contents of serum IL-6,TNF-αand MMP-9,and the ratios of p-JAK2/JAK2 and p-STAT3/STAT3,and the expression level of MMP-9 protein in knee cartilage tissue were significantly increased in the model group(P<0.01).In comparison with the model group,the Lequesne score,Mankin score,contents of serum IL-6,TNF-αand MMP-9,and the ratios of p-JAK2/JAK2 and p-STAT3/STAT3,and the expression of MMP-9

关 键 词：温针灸 膝关节骨关节炎 软骨 Janus蛋白酪氨酸激酶2/信号转导和转录活化因子3信号通路 炎性因子 

分 类 号：R245.8[医药卫生—针灸推拿学]