Objective To investigate the mechanism of action of Guyanxiao Fang on the regulation of autophagy in bone and joint cells through the PI3K/Akt/mTOR signaling pathway.Methods A chondrocyte inflammation model was constructed,and cells were randomly divided into 6 groups:blank group(rabbit chondrocytes + complete culture medium),model group(rabbit chondrocytes + 10 μg/mL LPS culture medium),positive group(rabbit chondrocytes + LPS + 150 μmol/L D-glucosamine),low-dose Guyanxiao group(rabbit chondrocytes + LPS + 1 μg/mL Guyanxiao),medium-dose Guyanxiao group(rabbit chondrocytes + LPS + 10 μg/mL Guyanxiao),and high-dose Guyanxiao group(rabbit chondrocytes + LPS + 100 μg/mL Guyanxiao).Cell proliferation was detected by the CCK-8 method;the levels of autophagy factors ATG5 and ATG12 in cell supernatants were detected by enzyme-linked immunosorbent assay;the relative expression levels of PI3K,Akt,and mTOR genes in cells were detected by real-time fluorescence quantitative polymerase chain reaction;Western blotting was used to detect the relative expression levels of PI3K,Akt,mTOR,and Beclin-1 proteins.Results The differences in cell viability among the blank group,model group,and different concentration groups were statistically significant(P<0.05).Compared with the blank group,the levels of ATG5 and ATG12 in chondrocytes of the model group,positive group,and high,medium,and low-dose Guyanxiao groups were all decreased(P<0.05);compared with the model group,the levels of ATG5 and ATG12 in chondrocytes of the positive group and high,medium,and low-dose Guyanxiao groups were all increased(P<0.05).Compared with the model group,the relative expression levels of PI3K,Akt,and mTOR mRNA in chondrocytes of the high,medium,and low-dose Guyanxiao groups were all decreased(P<0.05).Compared with the blank group,the relative expression levels of PI3K,Akt,and mTOR proteins in chondrocytes of the model group were all increased(P<0.05),while the relative expression level of Beclin-1 protein was decreased(P<0.05);compared with the model group,the relative expression levels of PI3K,Akt,and mTOR proteins in chondrocytes of the positive group and high,medium-dose Guyanxiao groups were decreased(P<0.05),and the relative expression levels of Beclin-1 protein in chondrocytes of the positive group and high,medium,and low-dose Guyanxiao groups were increased(P<0.05).Conclusion Guyanxiao Fang can effectively prevent and treat osteoarthritis.Its mechanism of action may be through inhibiting the PI3K/Akt/mTOR signaling pathway,causing autophagy to be regulated to a certain extent,promoting chondrocyte proliferation and differentiation,thereby slowing down the development of osteoarthritis.
维普
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