| ObjectiveIn this study,we observed the regulation of gene and protein expression of BMP-12,Smad1/5/8,Co L-I in rabbit behavior,histomorphology and ligament tissues by needle-knife in KOA treatment,to explore the underlying mechanism of needle-knife in promoting ligament healing and improving ligament properties in koa treatment,and to provide more experimental basis for needle-knife intervention in the treatment of KOA.MethodsThirty-two rabbits were randomly divided into a normal group,a model group,an electroacupuncture group,and a needle-knife group,and a KOA rabbit model was constructed by fixed brakes using the modified Videman method.The electroacupuncture group and the needle-knife group underwent a 3-week therapeutic intervention,respectively,and the electroacupuncture group selected four acupoints(Yanglingquan,yinglingquan,inner knee eye,and outer knee eye)for electroacupuncture treatment,and the needle-knife group performed a release maneuver on the inner and outer collateral ligaments and patellar ligaments from the middle segment back to and forth toward the starting point of the ligaments.Behavioral observation assessments were performed after modeling and after treatment;Gross structure and light microscopic organization of the knee joint cavity to observe the pathological appearance of knee ligaments in the affected limbs of rabbits;The levels of protein translation and gene transcription of BMP-12,Smad1/5/8,Co L-I in ligament tissues were determined by Western Blot analysis with real time PCR,respectively.The above indicators were statistically analyzed separately to observe the regulation effect of needle-knife on rabbits with knee osteoarthritis.Results1.Behavioral results: After modeling,the ratio of Lequesne MG score of rabbits in model group,electroacupuncture group and needle-knife group increased significantly compared with that of normal group(P<0.05).After treatment,the scores of both the electroacupuncture group and the needle-knife group decreased significantly,which was significantly different from the model group(P<0.05),and the needle-knife group had lower scores than the electroacupuncture group(P<0.05).In terms of score improvement,the experimental rabbits in the normal group had normal scores,and all the changes in scores compared with the other three groups were statistically significant(P<0.05).Compared with the model group,the electroacupuncture group and the needle-knife group showed greater improvement in the integral scores(P<0.05).Significant differences(P<0.05)were observed between the electroacupuncture group and the needle-knife group.2.Histomorphometric results: Both gross observation of the joint cavity and light microscopic observation of the ligament tissue could show that the pathological condition of KOA in the model group was obvious,as the joints were swollen and congested,the ligament cords were dull and thin,and had adhesions to the surrounding tissues,the synovium appeared hyperplastic,the joint fluid was turbid and increased in amount,and the articular cartilage was defective and rough,color and dark,and osteophyte formation were observed,microscopically,the collagen fibrils were loose or fractured,their arrangement was disordered,the bundles of normal collagen fibrils disappeared,and the fibroblast nuclei were arranged in a disorganized and significantly increased in number,with altered morphology.The above manifestations were alleviated to varying degrees in both the electroacupuncture and needle-knife groups,but the changes were more obvious in the needle-knife group than in the electroacupuncture group.3.Western Blot results: Compared with the normal group,the protein expressions of BMP-12,Smad1/5/8,and Co L-I in the knee ligaments of rabbits in the model group were significantly decreased(P<0.05).Compared with the model group,the contents of BMP-12,Smad1/5/8,and Co L-I in the electroacupuncture group and the needle-knife group were up-regulated(P<0.05).Compared with the electroacupuncture group,an increasing trend was observed in the needle-knife group in terms of the expression level of Co L-I(P<0.05),but there was no significant difference in the expression of BMP-12,Smad1/5/8(P>0.05).4.Real-time PCR results: Compared with the normal group,the m RNA expression levels of BMP-12 and Co L-I in the knee ligaments of the affected limbs of the rabbits in the model group were significantly decreased(P<0.05).Compared with the model group,the expression of BMP-12 and Co L-I m RNA content in the electroacupuncture group was up-regulated overall compared with that in the needle-knife group(P<0.05).Compared with the electroacupuncture,group,an increasing trend(P<0.05)was observed in terms of the expression level of Co L-I gene in the ligament of the needle-knife group,but the difference in the transcription level of BMP-12 gene was not statistically significant(P>0.05).There were no significant differences between the two groups in the expression levels of Smad1/5/8 m RNA content in the ligaments of normal,model,electroacupuncture,and needle-knife groups(P>0.05).Conclusion1.Electroacupuncture and needle-knife intervention can improve the general condition,behavior,histomorphology and expression of related factors in ligaments of KOA rabbit model,and needle-knife intervention has the better effect.2.The intervention of needle-knife can activate BMP-12/Smad signaling pathway to stimulate the generation and differentiation of tendon cells in ligament tissues,promote the endogenous healing of ligaments and correct the mechanical environment of the knee joint.3.The intervention effect of needle-knife could promote the generation of Co L-I to reinforce the mechanical properties of collagen fibers and fiber bundles,optimize the survival environment of chondrocytes. |
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