Effect Of Aconitum Injection On Sox-9 In Osteoarthritis Joint Fluid Of Rabbit Knee

OBJECTIVE: To observe the changes of articular cartilage tissue of rabbits in each group by naked eye and microscope,and to detect the content of Sox-9 in articular fluid by enzyme-linked immunosorbent assay(ELISA),the aim of this study is to explore the pathogenesis of Aconitum osteoarthritis/osteodystrophy through the direction of tissue and molecular signal pathway,and to analyze the changes of various indexes,so as to provide a strong theoretical basis for clinical use of aconitum injection and explore new ways of drug use.METHODS: thirty-two Healthy five-month-old New Zealand white rabbits were randomly assigned to a group of 7 blank rabbits and a model group of 25 rabbits after one week of adaptive feeding.To mold a mold set.In order to ensure the validity of the model,one of the two groups was selected at random and dissected to observe the pathological changes of knee joint(pre-experiment).Then 24 rabbits were randomly divided into model group(B),sodium hyaluronate control group(C),Zhengqing fengtongning control group(d)and aconitum injection treatment group(e).Then every three days(I.E.Day 1,4,7,10),double posterior knee joint cavity drug injection,a total of four injections,Group C was given 0.2ml sodium hyaluronate injection,group D was given 0.2 ml Zhengqing fengtongning injection,group E was given 0.2 ml aconite injection,group B was given 0.2 ml normal saline,group a was not given any treatment.All 30 rabbits were killed by air embolism method.The knee joint fluid of each rabbit was collected,centrifuged,refrigerated and kept for a week.The articular capsule was opened to observe the articular cartilage of the knee joint,and the image data were collected.Experiment Items: 1.Gross Observation of pathological joints;2.Morphology under light microscope;3.enzyme-linked immunosorbent assay for Sox-9 content in samples.RESULTS:(1)By observing the photographs of the articular capsule,we can see that:Group A is normal;Group B has the most serious injury of knee joint,the local changes are great,the cartilage surface is rough and the excrescent bone is formed;Group C and D Have a little degeneration;Group E is the closest to group A.(2)Under the light microscope,the observation results of the articular cartilage slices showed that the cells in group a were normal,the pathological changes in Group B were obvious,the cartilage cells in groups C and d were slightly overlapped and disordered,and the chondrocytes in Group E were basically normal,by comparison,it's closest to group A.(3)According to strict Mankin's score,Group A had the lowest score,Group B had the highest score and suffered the most serious injury,and Group C and D had slightly higher scores Than Group A,which indicated that the scores of group a were significantly lower after treatment There was no significant difference between Group E and Group A.(4)The expression of Sox-9 in synovial fluid was the highest in Group A and the lowest in Group B,and the expression of Sox-9 in Group C and D was the highest in Group A and the lowest in Group B,the protein concentration of the two groups was higher than that of Group B,but there was no significant difference between the two groups.CONCLUSION: 1.Through the direct observation and grading of the knee joint surface of rabbits,it was found that aconitum injection,Zhengqing fengtongning and sodium hyaluronate all had a certain therapeutic effect on knee joint osteoarthritis/osteodystrophy,and Aconitum injection had better effect;2.Through microscopic observation,we can see: ACONITUM injection group significantly decreased Mankin's score,so it can effectively protect the knee chondrocytes,maintain the normal order of Chondrocytes;3.Elisa results: aconitum injection can inhibit the initiation of Wnt /catenin classical pathway,block the signal factor to the downstream,and increase the expression of Sox-9 and aggrecan,promote the chondrocyte differentiation and protect the articular cartilage;The expression of Sox-9 and Aggrecan was positively correlated to some extent,and regulated the expression of Type II Collagen together.