Study on the Effect of Zhang's Chubi Recipe on Inflammation and Apoptosis of Rat Degenerative Chondrocytes by Regulating miR-34a/SIRT1 Signaling Axis
Objective:To study the effect and mechanism of Zhang's Chubi Recipe on inflammation and apoptosis in degenerative chondrocytes of rats. Methods:Rat articular cartilage T25 cells were taken and divided into control group,model group,CBF group,CBF+miR-34 a mimic group,and CBF+miR-34 a mimic+SIRT1 group. IL-1β was used to establish a degenerative chondrocyte model in the model group and CBF group. The CBF+miR-34 a mimic group was transfected with miR-34 a mimic and treated with IL-1β. The CBF+miR-34 a mimic+SIRT1 group was co-transfected with overexpression plasmids SIRT1 and miR-34 a mimic. And treated with IL-1β. CBF group,CBF+miR-34 a mimic group and CBF+miR-34 a mimic+SIRT1 group were added with Zhang's Chubi recipe containing serum. The level of miR-34 a in T25 cells was determined by real-time quantitative PCR. The levels of ALP,BGP,SOD,MDA,GSH,IL-6 and TNF-α were determined by ELISA kit. The apoptosis rate was determined by flow cytometry. The level of SIRT1 protein in T25 cells was determined by Western blotting. The dual luciferase reporter assay was used to verify the targeting relationship between miR-34 a and SIRT1. Results:Compared with model group,the levels of miR-34a,MDA,IL-6,TNF-α,apoptosis rate of chondrocytes were decreased in CBF group (P<0.05),and the protein levels of SOD,GSH,ALP,BGP,SIRT1 were increased in chondrocytes (P<0.05). The results of double luciferase report showed that the relative activity of SIRT1 WT luciferase in miR-34a mimic group was decreased than that in miR-34a NC group (P<0.05). Conclusion:Zhang's Chubi recipe can significantly inhibit the inflammatory response of rat degenerative chondrocytes,repair oxidative stress injury,and inhibit chondrocyte apoptosis. The mechanism may be related to the regulation of miR-34a/SIRT1 signal axis.
Zhang's Chubi recipemiR-34a/SIRT1 signal axisDegenerative cartilageApoptosisInflammatory response
万方数据
维普
