| ObjectiveAcupotomy method was used to loosen the adhesion between the knee joint ligaments and the surrounding tissues to observe the morphological changes and the expression of TGF-β1,Col-Ⅲ,MMP-9 genes and proteins in rabbit ligaments of knee osteoarthritis model under the intervention of acupotomy Based on the molecular biology theory,explore the mechanism of acupotomy on the ligament tissue of the rabbit model of knee osteoarthritis,further explain the recovery effect of the intervention of acupuncture on the rabbit model of knee osteoarthritis,and provide a theory for the pathogenesis of knee osteoarthritis.in accordance with.MethodsThirty-two Japanese white rabbits,half male and half male,were randomly divided into a blank group,a model group,an electroacupuncture group,and a needle knife group,with 8 rabbits in each group.The left hind limbs were stretched and immobilized for 6weeks to make the model.After 6 weeks,the experimental rabbits were scored in behavior to analyze whether the model was successful or not.After the model was successfully made,the rabbits were raised for 1 week for treatment.The blank group and the model group received no treatment,and the electro-acupuncture group and the acupotomy group received intervention treatment.Both groups were treated for 3 weeks.Three weeks later,the morphology of the cartilage in the articular cavity of the rabbits was scored by the Jessica Badendick scale;the ligament tissues of each group of rabbits were taken,and the morphological changes of the ligaments were observed under light microscope;q RT-PCR and Western blot were used to detect the knee ligament tissues The expression of TGF-β1,Col-Ⅲ,MMP-9 m RNA and protein.Results1.General conditions: The rabbits in each group were eating,drinking,looking,and moving normally before making the model.They had energetic eyes and were sensitive to changes in the surrounding environment.After modeling,compared with the blank group,the rabbits in the model group,the electroacupuncture group and the needle-knife group reduced their diet and drinking water,looked sad,lacked energy,had less eyesight,were slow to respond to changes in the surrounding environment,and reduced their activity due to the restraint of their left limbs.The right limb was weight-bearing,and some rabbits had severe hair loss,loose stools and no food.After modeling,the rabbits in the electro-acupuncture group and the needle-knife group improved in their diet,drinking water,and expression,and the above changes gradually returned to normal.After 3 weeks of treatment,the knee joint activities of the rabbits in the electroacupuncture group and the needle-knife group improved,the amount of activity increased,and the swelling around the joints was significantly improved.The rabbits in the model group did not improve significantly in all aspects.2.Results of articular cartilage morphology score: Compared with the blank group,the rabbit Jessica Badendick score of the model group was significantly increased,and the difference was statistically significant(P<0.01);compared with the model group,the scores of the electroacupuncture group and the needle knife group were significantly reduced,which was significant Difference(P<0.01);Compared with the electro-acupuncture group,the score of the acupotomy group was lower,and the difference was statistically significant(P<0.05).3.Observation by light microscope: the rabbit knee ligaments in the blank group have complete tissue structure,with a large number of ligament cells and neat arrangement;the rabbit ligament structure in the model group is unclear,the ligament layer is thin and rough,the number of ligament cells is small,and the arrangement is scattered;After needle knife intervention,compared with the model group,the ligament structure of the big-ear white rabbit is clear,with more ligament cells,but the local cell arrangement is scattered,which is not as good as the blank group.4.Experimental results of q RT-PCR method: Compared with the blank group,the expression level of TGF-β1 and Col-Ⅲ m RNA in the model group was significantly decreased,and the expression level of MMP-9m RNA was significantly increased,with significant statistical difference(P<0.01);the electroacupuncture group and the model group There was no significant difference in the expression level of TGF-β1(P>0.05),the expression level of Col-Ⅲ m RNA increased,and the expression level of MMP-9m RNA decreased,and there was a significant statistical difference(P<0.05);comparison of TGF-β1 between the needle knife group and the model group,Col-Ⅲm RNA expression level increased,MMP-9m RNA expression level decreased,the difference was statistically significant(P<0.05);Compared with the electroacupuncture group,the Col-Ⅲ m RNA expression level had no significant difference(P>0.05)The expression level of TGF-β1m RNA was higher,and the expression level of MMP-9 m RNA was lower,and the difference was statistically significant(P<0.05).5.Western blot experiment results: Compared with the blank group,the expression of TGF-β1 and Col-Ⅲ protein in the model group decreased,and the expression of MMP-9protein increased,with significant statistical difference(P<0.01);compared with the model group,electroacupuncture The expression of TGF-β1 and Col-Ⅲ protein increased in the acupuncture group and the acupuncture group,while the expression of MMP-9 protein decreased,and the difference was statistically significant(P<0.05 or P<0.01);compared with the electroacupuncture group,the Col-Ⅲ protein in the acupuncture group There was no significant difference in expression(P>0.05),TGF-β1 protein expression was higher,MMP-9 protein expression was lower,and the difference was statistically significant(P<0.05).Conclusion1.Needle knife treatment can reduce the degeneration of articular cartilage and prevent the development of osteoarthritis from further expansion.2.Needle-knife treatment can reduce the damage caused by the adhesion of ligaments and surrounding soft tissues,significantly improve the ligament tissue,and restore the normal movement function of the knee joint.3.Needle-knife intervention effectively regulates m RNA and the protein expression levels of TGF-β1,Col-Ⅲ,MMP-9 in knee osteoarthritis ligament tissue,and protects the integrity of ligament tissue and articular cartilage.It may be one of the mechanism of action of needle-knife treatment of KOA One. |
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