The Effect Of Internal Heat-type Acupuncture Needle On The Expression Of Type Ⅰ Collagen,matrix Metalloproteinase-3 And Osteopontin In The Subchondral Bone Of Rabbit Knee Osteoarthritis Model

Objective To explore the effect of internal heat-type acupuncture needle on rabbit with knee osteoarthritis(KOA)on the expression of Collagen 1(COL-1),metalloproteinases-3(MMP-3),Osteopontin(OPN)in subchondral bone and learn about the possible mechanism of action.Methods 1.Groups and Animal model preparation A total of 30 New Zealand rabbits(specific-pathogen-free level)were divided into the blank group,the model group and the treatment group by random digital table,with 10 rabbits in each group.Rabbit KOA model was made by referring to Hulth method.2.Intervention The modified Lequesne MG scale was used to evaluate the behavior of rabbits in each group After 4 weeks,the treatment group were selected for the intervention of internal heat-type acupuncture needle on the left hind limb knee joint,once a week,for 20 minutes each time.The blank group and the model group received no intervention.3.Observation Indexes Toluidine Blue staining(TBS)was used to observe the morphological changes of subchondral bone tissue of rabbits in each group.Western blot(WB)and real-time quantitative PCR(q RT-PCR)were used to detect the expressions of COL-1,MMP-3,OPN protein and m RNA in the subchondral bone of the three groupsResults 1.Compared with the blank group before intervention,behavioral scores of model group and treatment group were significantly increased(P<0.05).Compared with the model group after intervention,the difference of behavioral evaluation between groups in the treatment group changed significantly,with statistical difference(P <0.05).2.Compared with the blank group,the percentage of subchondral bone trabecular area,trabecular thickness and trabecular number in the model group were significantly decreased(P <0.05),while trabecular spacing was significantly increased(P <0.05).Compared with the model group,the percentage of trabecular area and trabecular number in the treatment group were significantly increased(P<0.05),while trabecular spacing was significantly decreased(P<0.05),and the change of trabecular thickness was not significant.3.Compared with the blank group,The protein and m RNA expression levels of COL-1were significantly decreased(P <0.001),while the protein and m RNA expression levels of MMP-3,OPN were significantly increased(P <0.001).Compared with the model group,The protein and m RNA expression levels of COL-1 were significantly increased(P <0.05),while the protein and m RNA expression levels of MMP-3,OPN were significantly decreased(P<0.05).4.The change of COL-1 content was positively correlated with the percentage of trabecular area and the trabecular number,but negatively correlated with trabecular spacing.the change of OPN,MMP-3 and OPN/MMP-3 ratio was negatively correlated with the percentage of trabecular area and trabecular number,and positively correlated with trabecular spacing.Conclusion Internal heat-type acupuncture needle can improve the behavioral changes of KOA and subchondral bone microstructure in rabbits,which may be realized by up-regulating the expression of col-1 and down-regulating the expression of MMP-3 and OPN.