Effect Of Ultrasound Combined With Moxibustion On VEGF,MMP-9,TIMP-1 In Cartilage,synovial Membrane And Synovial Of Rabbit Knee Osteoarthritis

Research purposesTo study the effect of ultrasound combined with moxibustion on the rabbit knee osteoarthritis(KOA)model by observing the morpholog of articular cartilage and synovial membraney,and the expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)in cartilage,synovial membrane and synovial to define the efficacy and mechanism of ultrasound combined with moxibustion in the treatment of KOA.Research methods1 The establishment of animal models of KOA Thirty six-month-old New Zealand white rabbits were randomly divided into the operation group and the normal group(group E),the operation group were 24,and the normal group were 6.Except the E group,the operation group animal right knee were received anterior cruciate ligament transection(ACLT)to create a rabbit KOA model.2 Experimental animal grouping the operation group were randomly divided into ultrasonic combined with moxibustion group(group A),moxibustion group(group B),ultrasonic group(group C)and model group(group D),each group had 6 rabbits.3 Experimental intervention 6 weeks after operation,except group D and group E,ultrasound combined with moxibustion in group A,moxibustion treatment in group B,and ultrasonic treatment in group C.4 Experimental material After 2 weeks of treatment,animals were sacrificed by air embolism.Samples were collected to visually observe the approximate appearance of cartilage and synovial membranes.5 Index detection Morphology of synovium and cartilage was observed under HE light microscope,expression of VEGF,MMP-9 and TIMP-1 in cartilage and synovial membrane was detected by immunohistochenistry.Western blotting(WB)was used to detect the protein expression of MMP-9 in cartilage,and VEGF,MMP-9 and TIMP-1 in synovial fluid of each group.Real-time Quantitative Polymerase Chain Reaction(RT-qPCR)was used to detect the expression of VEGF mRNA,MMP-9 mRNA in cartilage,and VEGF mRNA in synovial fluid.Research results1 Gross observation of cartilage and synovial morphology:Compared with E group,A group,B group,C group and D group showed varying degrees of osteoarticular inflammation and synovial inflammation.A group,B group and C group of bone and joint inflammation and synovial inflammation was lighter than D group.2 Results of cartilage and synovial HE staining:The scores of cartilage and synovial membrane of A group,B group,C group and D group were higher than E group,the differences were statistically significant(P<0.05).The scores of cartilage and synovial membrane of A group,B group and C group were lower than those in D group,the differences were statistically significant(P<0.05).There were no significant difference between the scores of cartilage and synovial membrane of A group,B group and C group(P>0.05).3 Comparison of Chondrocytes and synovial cells of indicator optic density(IOD)values:Compared with E group,the IOD values of VEGF,MMP-9,TIMP-1 of cartilage and synovial of A group,B group,C group and D group were significantly increased,the differences were statistically significant(P<0.05).Compared with D group,the IOD values of VEGF,MMP-9,TIMP-1 of cartilage and synovial of A group,B group and C group were decreased,the differences were statistically significant(P<0.05).There were no significant difference between the IOD values of VEGF,MMP-9,TIMP-1 of cartilage and MMP-9,TIMP-1 of synovial membrane of A group,B group and C group(P>0.05).The IOD values of VEGF in cartilage of A group,B group and C group were not significantly different(P>0.05).The IOD values of VEGF in cartilage of A group and B group,B group and C group group were statistically significant(P<0.05).4 WB test results:Compared with E group,the expression of protein MMP-9 in cartilage and VEGF,MMP-9,TIMP-1 in synovial fluid of A group,B group,C group and D group were increased,there are differences Statistical significance(P<0.05).Compared with E group,the expression of protein MMP-9 in cartilage and VEGF,MMP-9,TIMP-1 in synovial fluid of A group,B group and C group were decreased,and the differences were significant(P<0.05).There was no significant difference between the expression of protein MMP-9 in cartilage and VEGF,MMP-9,TIMP-1 in synovial fluid of A group,B group and C group(P>0.05).5 RT-PCR results:Compared with E group,the expression of MMP-9mRNA and VEGFmRNA in cartilage of A group,B group,C group and D group were increased,the difference was statistical significance(P<0.05).Compared with D group,the expression of MMP-9mRNA and VEGFmRNA in cartilage of A group,B group and C group were lower than that of D group,and the differences were statistically significant(P<0.05).There was no significant difference in the expression of MMP-9mRNA and VEGFmRNA in cartilage between A group,B group and C group(P>0.05).Compared with E group,the expression of VEGF mRNA in synovial fluid of A group,B group,C group and D group were significantly increased(P<0.05).Compared with D group,the expression of VEGF mRNA in the synovial fluid of A group,B group and C group were significantly decreased(P<0.05).The expression of VEGF mRNA in the synovial fluid of C was lower than B group,the difference was statistically significant(P<0.05).The expression of VEGF mRNA in the synovial fluid of A group was lower than that of the C group,and the difference was statistically significant(P<0.05).The expression of VEGF mRNA in the A group was lower than that of the B group,and the difference was statistically significant(P<0.05).Research conclusion1 Ultrasound combined with moxibustion,moxibustion and ultrasonic treatment can delay KOA cartilage degeneration and reduce synovial inflammation in rabbits.2 There is no obvious difference among ultrasound combined with moxibustion,moxibustion and ultrasound to treat rabbit KOA.However,the combination of the two may have different effects on different tissues of the joint,which is different from simple ultrasound or moxibustion alone.It may be simple to choose a method of ultrasonic or moxibustion,without a combination of various physiotherapy methods in Physical factor therapy of Chinese and Western Medicine to treat KOA in the clinical.The method can provide certain reference for treating KOA,but it still needs further study in the future.3 Ultrasound group and moxibustion group may be used to treat rabbit KOA by inhibiting the expression of VEGF,MMP-9,TIMP-1 in cartilage,synovial membrane and synovial to delay the degeneration of cartilage and reduce the inflammation of synovial membrane and synovial,playing a role in the treatment of rabbit KOA.