Effects Of Modified Duhuo Jisheng Mixture On Type II Collagen And OPG/RANKL Axis In Cartilage Of Rabbits With Knee Osteoarthriti

Objective: To observe the effect of Jiawei Duhuo Jisheng mixture on the Collagen Ⅱ and OPG/RANKL level and Collagen Ⅱ expression in rabbit KOA model cartilage,and to enrich the mechanism of action of Jiawei Duhuo Jisheng Mixture in treating knee osteoarthritis.Methods: 24 3-month-old male New Zealand rabbits were randomly divided into a blank group,a model group,a Jiawei Duhuo group and a Celecoxib group,with 6 rabbits in each group.The early KOA model was established by injecting type Ⅱ Collagenase into the knee joint.After 2weeks,measure the skin temperature,circumference and gait score of each group of rabbits,and preliminarily test whether the KOA model was successful.Give the medication for another 6weeks.At the 9th week,changes in cartilage morphology and structure in each group were observed using Safranin O-staining technique,and the protein levels of OPG,RANKL,and Collagen Ⅱ in each group’s cartilage were detected using immunofluorescence technique.The qPCR method was used to detect the mRNA expression levels of OPG,RANKL,and type Collagen Ⅱ.Results:1.the skin temperature,circumference and gait scores of rabbits in the Celecoxib group,the Jiawei Duhuo group and the model group were significantly higher than those in the blank group(P<0.01);2.The Safranin O-staining technique showed significant cartilage damage in the model group,and the Mankin’s score was higher than that of the blank group,with a statistically significant difference(P<0.01).Compared with the model group,the Mankin’s score of Celecoxib group and Jiawei Duhuo group decreased significantly(P<0.01).Compared with the Celecoxib group,there was no significant difference between the Jiawei Duhuo group and the Celecoxib group(P>0.05);3.The immunofluorescence results showed that compared with the model group,the expression of type Ⅱ collagen in the Jiawei Duhuo group was higher than that in the model group,with significant statistical significance(P<0.01).Compared with the blank group,the difference was not statistically significant(P>0.05).The mean optical density of OPG/RANKL in the Jiawei Duhuo group was the highest,which was significantly different from that in the Celecoxib group and the model group(P<0.01),and there was no significant difference compared with the blank group(P>0.05);4.qPCR detection showed that the OPGmRNA level in the Jiawei Duhuo group was significantly higher than that in other groups(P<0.01),and the RANKL level of Celecoxib was significantly higher than that in other groups(P<0.01).The OPG/RANKL of the bone joint surface in the Jiawei Duhuo group was significantly upregulated compared to other groups(P<0.01).The level of type Ⅱ collagen mRNA in the Jiawei Duhuo group was significantly higher than that in the model group(P <0.01),and higher than that in the Celecoxib group(P <0.05).Compared with the blank group,there was no significant difference(P >0.05).Conclusion:1.Injecting Type Ⅱ Collagenase into the knee joint can successfully establish rabbit KOA model,2.Jiawei Duhuo Jisheng Mixture can effectively promote cartilage repairing in KOA rabbits and delay the progression of KOA.Its mechanism of action may be related to upregulating type Ⅱ collagen and OPG/RANKL axis.