Mechanism Of Bushen Zhuangjin Decoction On The Inhibition Of Inflammation Mediated Cartilage Matrix Degradation

ObjectiveTo study the mechanism of Bushen Zhuangjin Decoction in inhibiting the degradation of cartilage matrix mediated by inflammationMethods1.A total of 45 rats of 2 month old SPF grade SD rats were selected,and the selected rats were divided into three groups by random number method,namely,the blank group(15)and the model group(30 rats).The blank group was treated with the false operation(physiological saline irrigation).The model group was established by improved Hulth method,and the model group was divided into the model group with the digital table method(15 rats,physiology).The rats were fed with salted water for stomach and Bushen Zhuang Jin Decoction group(15 rats,intragastric administration of Bushen Zhuang Jin Decoction),gavage 1 times per day,and continuous gavage for 12 weeks.The changes of chondrocyte arrangement,tidal line and bone trabecula were observed by HE staining.The number of chondrocytes and the matrix protein polysaccharide in articular cartilage were observed by toluidine blue staining.The expression of IL-1β、TNF-α and MMP-13 in synovial tissue was detected by ELISA.2.The intervention concentration of Bushen Zhuang Jin decoction was determined to be 200 ug/mL.The expression of IL-1β、TNF-a in supernatant of chondrocytes was higher in model group than in blank group(P<0.05,P<0.01),while that in Bushen Zhuang Jin Decoction group was lower than that in model group(P<0.05,P<0.01).The expression of MMP-3,MMP-13,ADAMTS4,AD AMTS 5 and RAF protein in the chondrocytes was higher than that in the blank group(P<0.05,P<0.01),and the group of tonifying kidney and Zhuang Jin Decoction group was lower than that of the model group(P<0.05).The expression of MMP-3,MMP-13,ADAMTS4,ADAMTS5 and RAF genes in the chondrocytes was higher than that in the blank group(P<0.05,P<0.01),and the group of Bushen Zhuang Jin Decoction group was lower than that of the model group(P<0.05,P<0.01).Results1.HE staining showed that the normal articular cartilage was divided into four layers:surface layer,migrating layer,radiant layer and calcification layer.The structure was clear and arranged,the tide line was clearly visible,the cartilage surface was smooth and complete,the cartilage cells were round and evenly distributed,arranged neatly,and the cytoplasm of the subchondral bone and the nucleus staining were deeper than the cartilage cells,arranged regularly.The width and density of the trabecular bone were moderate.The four layer structure of the model control group was not clear,the cartilage surface was rough,the shape was irregular,the cartilage thinned,the chondrocytes were uneven,the arrangement of the cartilage was disorderly,the tide line was blurred,twisted or interrupted;the color of the subchondral bone was difficult to distinguish between the cartilage layer,the cystic change,the sparsity of bone trabecula,The changes of cartilage and subchondral bone in Bushen Zhuang Jin Decoction group were between normal group and model group.Toluidine blue staining showed that the structure of normal articular cartilage was clear,the nuclei and cartilage matrix were stained clearly,and cytoplasm was almost unstained.In the model group,the color of cartilage matrix became shallow,the cartilaginous layer thinned,the cartilage cells became less and uneven,suggesting that the proteoglycan in the cartilage matrix was reduced and some of the cartilage cells were necrotic.The ELISA method was used to determine the expression of IL-1β、TNF-α、and MMP-13 in the synovium of the knee joint of rats.The results showed that the model group was significantly higher than that in the blank group(P<0.05,P<0.01),and the tonifying kidney Zhuang Decoction group was significantly lower than the model group(P<0.05,P<0.01),and there was no significant difference between the group of Tonifying the kidney and Zhuang Jin Decoction group and the blank group(P>0.05).The quantity of proteoglycan and chondrocytes in the cartilage matrix of the Bushen Zhuang Jin Decoction group was significantly better than that of the model control group.2.The concentration of Bushen zhuang jin decoction was 200 ug/mL with 8 h.The expression of IL-1β and TNF-a in the supernatant of chondrocytes was higher than that in the blank group(P<0.05,P<0.01),while those in the Bushen Zhuangjin Decoction group were lower than those in the model group(P<0.05,P<0.01).The expression of MMP-3,MMP-13,ADAMTS4,ADAMTS5 and RAF protein in the chondrocytes was higher than that in the blank group(P<0.05,P<0.01),and the Bushen zhuangjin decoction group was lower than that of the model group(P<0.05).The expression of MMP-3,MMP-13,ADAMTS4,ADAMTS5 and RAF genes in the chondrocytes was higher than that in the blank group(P<0.05,P<0.01),and the Bushen zhuangjin decoction group was lower than that of the model group(P<0.05,P<0.01).Conclusions1.By reducing the content of IL-1β、TNF-a and MMP-13 in synovial synovial membrane.Bushen Zhuang Jin Decoction inhibits the degradation of cartilage matrix mediated by inflammation.2.Bushen Zhuang Jin decoction can inhibit the expression of MMP-3,MMP-13,ADAMTS4.ADAMTS5 and RAF,inhibit LPS mediated inflammatory reaction,and maintain cartilage matrix homeostasis.