GJK Tablets intervene in cartilage homeostasis to protect articular cartilage of mice with knee osteoarthritis

Li Yijin; Li Jiahao; Zhang Haitao; Huang Yiwei; Chen Jinlun; Zeng Yirong; Feng Wenjun

doi:10.12307/2026.661

Abstract：BACKGROUND:Previous studies have found that GJK Tablets can improve joint pain and function in patients with knee osteoarthritis,but the specific mechanism remains unclear. OBJECTIVE:To explore the mechanism underlying the chondroprotective effect of GJK Tablets in a mouse model of knee osteoarthritis. METHODS:Network pharmacology was used to obtain the common targets of GJK Tablets and cartilage homeostasis,and a protein-protein interaction network was constructed.Key signaling pathways were screened through gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis.Experimental mice were randomly divided into a control group,a model group,low-,medium-,and high-dose GJK Tablet groups,and a positive drug group(glucosamine hydrochloride capsules).In the latter five groups,the modified Hulth method was used to establish the mouse model of knee osteoarthritis.In the control group,only the relevant tissues of mice were exposed but the ligaments and menisci were not removed.After modeling,the low,medium-,and high-dose groups were intragastrically administered with 187.5,375,and 750 mg/kg GJK Tablet suspension,respectively.The positive drug group was intragastrically administered with 187.5 mg/kg glucosamine hydrochloride suspension.The model group and the control group were intragastrically administered with an equal volume of normal saline.The administration was carried out every other day for 8 weeks.After the intragastric administration,the pathological morphology of the knee joint cartilage of the mice was observed by hematoxylin-eosin and safranin O-fast green staining,and the Osteoarthritis Research Society International(OARSI)score for the cartilage was evaluated.The protein expressions of SRY-Box transcription factor 9(Sox9),type II collagen,and matrix metalloproteinase 13 in the mouse cartilage tissue were detected by immunohistochemical staining. RESULTS AND CONCLUSION:(1)A total of 140 intersection targets of GJK Tablets and cartilage homeostasis were obtained,and Sox9 was one of the targets.(2)Gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that the main signaling pathways involved were cancer-related pathways,hypoxia-inducible factor 1,MAPK,PI3K/Akt,nuclear factor κB,transforming growth factor β and other signaling pathways.(3)The results of hematoxylin-eosin staining and safranin O-fast green staining showed that compared with the control group,the cartilage contour in the model group was deformed,the defect was stratified,the number of chondrocytes was significantly reduced,the tidemark was blurred,and the OARSI score was higher.Compared with the model group,the cartilage surface in the low-,medium-,and high-dose GJK Tablet groups was more intact,the cartilage thickness and the number of chondrocytes were increased,the tidemark was clearer,and the OARSI scores were significantly decreased.(4)The immunohistochemical results indicated that compared with the control group,the protein expressions of Sox9 and type II collagen in the model group were significantly decreased,and the expression of matrix metalloproteinase 13 was significantly increased(P<0.01).Compared with the model group,the expression of matrix metalloproteinase 13 in the low-,medium-,and high-dose GJK Tablet groups was significantly decreased(P<0.01).The positive expressions of Sox9 and type II collagen in the medium-and high-dose GJK Tablet groups were significantly increased(P<0.01),and the positive expressions showed an increasing trend from the low-dose group to the high-dose group(P<0.01).To conclude,GJK Tablets can intervene in cartilage homeostasis by regulating Sox9,playing a chondroprotective role in knee osteoarthritis.Signaling pathways such as hypoxia-inducible factor 1,MAPK,PI3K/Akt,nuclear factor κB,and transforming growth factor β may be the main pathways through which GJK Tablets intervene in cartilage homeostasis.