| Purpose: Observing the effects of Gan-Jiang-Ling-Zhu decoction(GJLZ)on the expression levels of transforming growth factor-β1(TGF-β1)and matrix metalloproteinase-3(MMP-3)in the joint fluid of rabbit Knee Osteoarthritis(KOA)model.This experiment further deepened the understanding of the changes in the course of KOA,discussed the pharmacological effects of GJLZ decoction,and provided new solutions for the clinical treatment of KOA.Material and method: Twenty eight Japanese white rabbits were randomly divided into four groups: blank group,model group,diclofenac sodium group and GJLZ group.There were 7 rabbits in each group,and the blank group was not treated.The model group,GJLZ group and diclofenac sodium group used the improved Hulth method to prepare the KOA model rabbits,choosing the left posterior knee joint.Two weeks after modeling,a Japanese white rabbit was randomly selected in each group,and the modeling joint was observed after being killed.The modeling was evaluated in combination with the Lequesne MG index assessment,and the operation point was observed and recorded.After successful modeling,drug intervention treatment was carried out according to the experimental design.Each of the blank group and the model group was given 5ml/d distilled water by gavage;In GJLZ group,each rabbit was given a conventional Decoction of 4g,5ml/d by gavage;In diclofenac sodium group,8.3mg diclofenac sodium was dissolved in 5ml/d distilled water by gavage;The Japanese white rabbits in each group were given continuous intragastric administration for 4weeks.During the treatment,the behavioral changes of Japanese white rabbits were observed and recorded with Lequesne MG score as the index.After 4 weeks of drug intervention,the knee cartilage of each group was observed,and the morphological changes of each group was observed under light microscope after Hematoxylin-Eosin(HE)staining.The protein contents of a a transforming growth factor-β1(TGF-β1)and matrix metalloproteinase-3(MMP-3)nd B in articular fluid were detected by ELISA and Western blot.Results:1.GJLZ decoction could significantly improve the behavioral score of experimental animals.Lequesne MG index assessment was normal before modeling.After modeling,the scores of model group,GJLZ Decoction group and diclofenac sodium group were slightly higher,but there was no significant difference between groups(P > 0.05).2.Morphological observation of cartilage after Hematoxylin-Eosin(HE)staining.Compared with the model group,the surface cartilage exfoliation of GJLZ Decoction group and diclofenac sodium group was less,the clustering performance was not obvious,and the nuclear degeneration was slight,and the improvement degree of GJLZ Decoction group was slightly better than that of diclofenac sodium group.3.Western blot showed that compared with the blank group,the expression levels of MMP-3and TGF-β1 in rabbit knee joint fluid in the other three groups increased,and the difference was statistically significant(P < 0.05);Compared with diclofenac sodium group,the expression levels of MMP-3 and TGF-β1 in rabbit knee joint fluid in GJLZ decoction group decreased,and the difference was statistically significant(P < 0.05).4.ELISA showed that compared with the blank group,the expression levels of MMP-3 and TGF-β1 in rabbit knee joint fluid in the other three groups increased,and the difference was statistically significant(P < 0.05);Compared with diclofenac sodium group,the expression levels of MMP-3 and TGF-β1 in rabbit knee joint fluid in GJLZ decoction group decreased,and the difference was statistically significant(P < 0.05);Conclusion:1.The improved Hulth method used in this experiment can replicate the KOA model of Japanese big ear white rabbits,which can make the model rabbits show the characteristics of arthritis such as joint pain,swelling and limited activity,and the two inflammatory factors MMP-3 and TGF-β1 in joint fluid are increased.It can be effectively modeled,with the characteristics of fast modeling,low cost and good stability.2.After GJLZ decoction intervened KOA rabbits,through the observation and scoring of behavior and cartilage morphology,it is suggested that this prescription can effectively increase the range of motion of knee joint,reducing the symptoms of pain and swelling,the degree of synovitis thickening,and improve the damage of articular cartilage to a certain extent.3.The therapeutic mechanism of GJLZ decoction on rabbit knee cartilage injury is closely related to its ability to reduce the expression levels of MMP-3 and TGF-β1 in knee joint fluid. |
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