Effect of Guilu Erxian Jiao on IL-1 β-induced AMPK/mTOR/ULK1 pathway media-ted autophagy in cultured chondrocytes in vitro
Objective Investigate the effect of Guilu Erxian Jiao on the autophagy of cultured chondrocytes mediated by the IL-1β induced AMPK/mTOR/ULK1 pathway.Methods Wild type mouse and mTOR-KO mouse chondrocytes were cultured in vitro by enzyme elimination method.Mouse chondrocytes were divided into blank group,model group,Guilu group,CKO group and Guilu+CKO group.After 24h of intervention,chondrocyte activity was detected by CCK-8 method and apoptosis was detected by TUNEL method;The expression of LC3B protein in chondrocytes of each group was observed by laser confocal microscope;Western blotting and qRT-PCR were used to detect the expression of mTOR,ULK1,AMPK,LC3B,ATG5,PARP-1 protein and mR-NA.Results Compared with blank group,the chondrocyte activity in model group was significantly decreased(P＜0.05),and the apoptosis rate of chondrocytes was significantly increased(P＜0.01);LC3B light intensity decreased significantly(P＜0.01);the protein and mRNA expressions of mTOR and PARP1 were increased(P＜0.05),and the protein and mRNA expres-sions of ULK1,AMPK,Atg5 and LC3B were decreased(P＜0.05).Compared with model group,the chondrocyte activity in tur-tle deer group was significantly increased(P＜0.05),and the apoptosis rate of chondrocytes was significantly decreased(P＜0.01);LC3B light intensity was significantly increased(P＜0.01);the protein and mRNA expressions of ULK1,AMPK,Atg5,LC3B were increased(P＜0.05),and the protein and mRNA expressions of mTOR and PARP1 were decreased(P＜0.05).Compared with turtle and deer group,the chondrocyte activity in CKO group was significantly increased(P＜0.05),and the ap-optosis rate of chondrocytes was significantly decreased(P＜0.01);LC3B light intensity was significantly increased(P＜0.01,P＜0.05);the protein and mRNA expressions of mTOR and PARP1 were decreased(P＜0.05),and the protein and mRNA ex-pressions of ULK1,AMPK,Atg5 and LC3B were increased(P＜0.05).Compared with CKO group,chondrocyte activity in tur-tle deer+CKO group was significantly increased(P＜0.01),and the apoptosis rate of chondrocytes was significantly decreased(P＜0.05);LC3B light intensity was significantly increased(P＜0.05);protein and mRNA expressions of ULK1,AMPK,Atg5,LC3B were increased(P＜0.05),protein and mRNA expressions of PARP1 were decreased(P＜0.05),and protein and mRNA expressions of mTOR were decreased.There was no significant difference(P＞0.05).Conclusion Guilu Erxian Jiao can induce autophagy by activating AMPK/mTOR/ULK1 pathway,promote the proliferation of degenerative chondrocytes,and then reduce the apoptosis of chondrocytes.
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