| ObjectiveCamparing before and after warming acupuncture treatment of rabbits knee osteoarthritis cartilage JNK1\2、 MMP-1\13 levels of expression, the warm acupuncture mechanism is be cleared in the treatment of the knee osteoarthritis.MethodOnly 40 New Zealand big-eared rabbits were randomly divided into the wraming acupuncture group, the acupuncture group, the model group and the blank group, making rabbit knee osteoarthritis with right unbend leg plaster fixation method. After the model is builded,the wraming acupuncture group and the acupuncture group must be treamtent by acupuncture Wai xiyan, Nei xiyan, He ding, once a day, 15 min once, a tolal of 2 weeks treatment. The model group and blank gruop have normal feeding without treatment. Collect the serum and cartilage after all treatment. To observe the knee joint morphology changes by Peng Taiping observation method. To test the front building, before and after treatment in rabbit serum expression level of MMP-1, MMP-13 by using double antibody sandwich enzyme-linked immunosorbent(ELISA) detection. Using Immunohistochemistry detection to observe the expression of MMP-1, MMP-13 in articular cartilage. To detect JNK1、 2 expression by Western Blot method. All the results were be statistical analysis using SPSS17.0 software.Result1.After 2 weeks treatment, observed the knee joint cartilage become thinning in model group, a significant articular cartilage injury. Warm acupuncture group can obviously improve the histological structure of articular cartilage.2. Hematoxylin-eosin staining results show that the cartilage surface is smooth, the structure is integrity, the cartilage cells is in alignment and shows four layers in blank group. The cartilage become thinning, and the number of cartilage cells is decreased, disordered arrangement in model group.The cartilage cells is incresed and the cartilage edge is smooth in warming acupuncture group.In the acupuncture group, the cartilage edge is less smooth.3.The expression of serum MMP-1 、 13 levels in ELISA: the expression of MMP-1 、 13 were significantly higher, as compared with that before model establishment(P<0.05). After treatment, the model group compared with blank control group, serum levels of MMP-1 、 13 were increased, the difference was statistically significant(P<0.05);The warming acupuncture group serum levels of MMP-1 、 13 expression significantly reduced, the difference was statistically significant(P<0.05), as compared with the model group. The acupuncture group had decreased, but there was no statistically significant difference(P>0.05)compared with model group.4.Positive results of immunohistochemical detection optical microscope MMP-1 and MMP-13 distribution: In the blank group, only in the positive expression of MMP-1 and MMP- 13, and the expression is not obvious; the model group cartilage cell coloring is relatively deep, mainly in the darker brown yellow brown particles as the main, and extracellular matrix are also a small amount of coloring; positive expression of the warming acupuncture group cartilage cell MMP-1, MMP-13 mainly in the shallow brown particles, surface slightly darkbrown particles distribution; the positive expression of Voltaren group is obvious,and the brown granules of surface layer and the middle deep color distribution of higher.5.The expression of cartilage MMP-1 、 13 levels in immunohistochemical detection: Comparing with blank control group, the expression of MMP-1、13 were increased significantly in the model group, the difference was statistically significant(P<0.05); Comparing with the model group, the expression of MMP-1and MMP-13 were significantly reduced in warming acupuncture group, the difference was statistically significant(P<0.05); Comparing with model group,there was no statistically significant difference(P>0.05).6.The expression of cartilage JNK1 、 2 levels in western Blot detection:Comparing with blank control group, the expression of JNK1 、 2 were increased significantly in the model group, the difference was statistically significant(P<0.05);Comparing with the model group, the expression of JNK1 、 2 were significantly reduced in warming acupuncture group, effectively inhibits JNK1 、 2expressing, the difference was statistically significant(P<0.05);Comparing with model group, there was no statistically significant difference(P>0.05).ConclusionThe warming acupuncture treatment of KOA may be through the activation of JNK signaling pathway and inhibition of MMP-1,MMP-13 expression, and give play to the role of the treatment mechanism. |
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