Effects of decoction of Tortoise Plastron Glue plus Deerhorn Glue and its decomposed on IL-1β-induced apoptosis and extracellular matrix of chondrocytes via Caspase-3/PARP pathway
Objective It is to investigate the effects of decoction of Tortoise Plastron Glue plus Deerhorn Glue and its decomposed on interleukin-1β(IL-1β)induced apoptosis and extracellular matrix(ECM)of chondrocytes via cysteine as-partic protease 3(Caspase-3)/poly ADP-ribose polymerase(PARP)pathway.Methods The blank serum,decoction of Tortoise Plastron Glue plus Deerhorn Glue containing serum,Tortoise Plastron Glue containing serum and Deerhorn Glue containing serum were prepared using SD rats,and the C57BL/6 mouse chondrocytes were cultured by enzyme ablation method in vitro.The chondrocytes were divided into 5 groups:blank group cultured with blank containing serum,model group cultured with IL-1β and blank containing serum,Tortoise Plastron Glue plus Deerhorn Glue group cultured with IL-1β and Tortoise Plastron Glue plus Deerhorn Glue containing serum,Tortoise Plastron Glue group cultured with IL-1β and Tortoise Plastron Glue containing serum,Deerhorn Glue group cultured with IL-1β and Deerhorn Glue containing serum.After 24h of intervention,the activity of chondrocyte was detected by CCK-8 method and its apoptosis was detected by TUNEL method,the expressions of PARP-1 and Caspase-3 were detected by immunofluorescence staining,the protein and mRNA expressions of PARP-1,Caspase-3,matrix Metallopeptidase 13(MMP-13),aggrecan were detected by Western blotting and qRT-PCR.Results Compared with the blank group,the activity of chondrocytes in the model group was signifi-cantly decreased(P＜0.05),and the apoptosis rate of chondrocytes was significantly increased(P＜0.05);the average fluorescence intensity of PARP-1 and Caspase-3,the protein and mRNA expressions of PARP-1,Caspase-3,MMP-13 in the cells were significantly increased(all P＜0.05),while the protein and mRNA expressions of Aggrecan were signifi-cantly decreased(all P＜0.05).Compared with the model group,the activities of chondrocytes in the Tortoise Plastron Glue plus Deerhorn Glue group,Tortoise Plastron Glue group,Deerhorn Glue group were significantly increased(all P＜0.05),and the apoptosis rates of chondrocytes were significantly decreased(all P＜0.05);the average fluorescence in-tensity of PARP-1 and Caspase-3,the protein and mRNA expressions of PARP-1,Caspase-3,MMP-13 in the cells were significantly decreased(all P＜0.05),while the protein and mRNA expressions of Aggrecan were significantly increased(all P＜0.05).Compared with the Tortoise Plastron Glue group and Deerhorn Glue group,the Tortoise Plastron Glue plus Deerhorn Glue group had higher chondrocyte activity(both P＜0.05)and lower chondrocyte apoptosis rate(both P＜0.05),lower average fluorescence intensity of Caspase-3 and PARP-1 and the relative expressions of Caspase-3,PARP-1,MMP-13 protein and mRNA in the cells(all P＜0.05),and higher cellular Aggrecan protein and mRNA relative expres-sion(all P＜0.05).Compared with the Tortoise Plastron Glue group,the activity of chondrocytes,apoptosis rate of chon-drocytes,mean fluorescence intensity of PARP-1 and Caspase-3 in the cells,protein and mRNA expressions of PARP-1,Caspase-3,MMP-13,Aggrecan in the cells were significantly higher in the Deerhorn Glue group(all P＜0.05).Conclu-sion Tortoise Plastron Glue can protect chondrocytes by inhibiting chondrocyte apoptosis and ECM degradation,and Deer-horn Glue can promote chondrocyte proliferation and improve the anabolism level of ECM,the effect of decoction of Tortoise Plastron Glue plus Deerhorn Glue is better than Tortoise Plastron Glue and Deerhorn Glue alone,the mechanism of action may be related to the activation of the Caspase-3/PARP signaling pathway.
decoction of Tortoise Plastron Glue plus Deerhorn Gluechondrocyteapoptosisextracellular matrixcysteine aspartic protease 3poly ADP-ribose polymerase
万方数据
维普
