温针灸对兔膝骨性关节炎模型软骨细胞中c-Jun氨基末端激酶信号通路的影响

Effect of Warming Moxibustion on Expression of c-Jun N-terminal Kinase of Cartilage Cells in Rabbits with Knee Osteoarthritis

目的观察温针灸治疗兔膝骨性关节炎(KOA)模型软骨中c-Jun氨基末端激酶(JNK)信号通路及基质金属蛋白酶(MMP)-1、MMP-13的表达,为温针灸治疗KOA提供现代医学理论支持。方法 40只新西兰大耳兔随机选择30只采用右膝关节伸直位石膏固定制备KOA模型,另10只为正常对照组。固定6周后行X线检查确定造模成功后随机分为温针灸组、双氯芬酸钠组、模型组,每组10只。温针灸组选取足三里、鹤顶、内膝眼穴给予温针灸治疗,每天1次,15min/次,治疗2周;双氯芬酸钠组按15mg/kg灌胃,每天1次,治疗2周;模型组及正常对照组不予任何干预,正常饲养。采用免疫组织化学法、RT-PCR、免疫蛋白印迹法(Western Blot)检测软骨中JNK1、JNK2、MMP-1、MMP-13的表达。结果与正常对照组比较,模型组关节软骨中MMP-1、MMP-13、JNK1、JNK2mRNA及蛋白表达升高(P<0.05),且温针灸组、双氯芬酸钠组关节软骨中MMP-1、MMP-13、JNK1、JNK2mRNA及蛋白表达较模型组降低(P<0.05),同时温针灸组MMP-1、MMP-13mRNA蛋白表达及JNK1、JNK2 mRNA表达较双氯芬酸钠组降低更为明显(P<0.05)。结论温针灸可抑制KOA软骨细胞中JNK1、JNK2、MMP-1、MMP-13的表达,温针灸治疗KOA可能是通过激活JNK信号通路,抑制MMP-1、MMP-13的表达而达到治疗作用。

Objective To observe the effect of the warming moxibustion（WM） on the expression of c-Jun N-terminal kinase（JNK） and matrix metalloproteinase-1/13（MMP-1 ,MMP-13） in knee cartilage ofosteoarthritis rabbits, and explore the mechanism of WM treatment on knee osteoarthritis（KOA）. Methods Ten rabbits were selected randomly from 40 rabbits as the normal controls group（NC group）, and the rest 30 rabbits were established model of KOA by extension position with plaster cast. After 6 weeks, X-ray checking was used to determine the model success. The 30 model rabbits were randomly divided into the WM group,the diclofenac sodium group（DS group） and the model group（MG group）, 10 in each group. WM at Zusanli （ST-36）, Heding （EX-LE2）, Neixiyan （EX-LE4） was performed in the WM group, 15 rain each, once per day. The rabbits in the DS group were administrated with diclofenac sodium by gavage 15 mg/kg, once per day for 2 weeks. The rabits in the MG and NC group received no treatment. The change of content of JNK1 ,JNK2,MMP-1 ,MMP-13 expression in cartilage after treatment were ob- served respectively by immunohistochemical method, reverse transcription-Polymerase Chain Reaction and Western Blot. Results Compared with the NC group,the mRNA and protein expressions of JNK1, J NK2,MMP-1, MMP-13 in the MG group were significantly increased （P 〈 0.05）. Compared with the MG group, the mRNA and protein expressions of JNK1 ,JNK2,MMP-1 ,MMP-13 in the WM and DS group were significantly decreased（P 〈0.05）. The mRNA and protein expressions of MMP-1 ,MMP-13 and the mRNA expression of JNK1, JNK2 in the MG group were decreased significantly than those in the DS group （P 〈 0.05）. Conclusions WM can inhibit the expression of J NK1, J NK2, MMP-1, MMP-13 in KOA chondro- cytes, indicating that WM might relives knee osteoarthritis by reducing the expression of MMP-1 and MMP-13 by JNK pathway.