摘要
SRT1720, a new discovered drug, was reported to activate silent information regulator 1（SIRT1） and inhibit the chondrocyte apoptosis. However, the underlying mechanism remains elusive. In the present study, the chondrocytes were extracted from the cartilage tissues of New Zealand white rabbits, cultured in the presence of sodium nitroprusside（SNP）（2.5 mmol/L） and divided into five groups： 1, 5, 10, and 20 μmol/L SRT1720 groups and blank control group（0 μmol/L SRT1720）. MTT assay was used to detect the chondrocyte viability and proliferation, and DAPI staining and flow cytometry to measure the chondrocyte apoptosis. The expression levels of SIRT1, p53, NF-κB/p65, Bax, and peroxisome proliferator-activated receptor gamma coactivator 1-α（PGC-1α） were detected by Western blotting and the expression levels of SIRT1, type Ⅱ collagen, and aggrecan m RNA by RT-PCR. The results showed that in the SRT1720-treated groups, the nuclei of chondrocytes were morphologically intact and had uniform chromatin. In the blank control group, nuclear rupture into debris was observed in chondrocytes. With the SRT1720 concentration increasing, the chondrocyte viability increased, the apoptosis rate decreased, the protein expression levels of SIRT1 and PGC-1α and the m RNA expression levels of type Ⅱ collagen and aggrecan increased（P〈0.05）, and the expression levels of p53, NF-κB and bax decreased（P〈0.05）. It was suggested that SRT1720 inhibits chondrocyte apoptosis by activating the expression of SIRT1 via p53/bax and NF-κB/PGC-1α pathways.
SRT1720, a new discovered drug, was reported to activate silent information regulator 1（SIRT1） and inhibit the chondrocyte apoptosis. However, the underlying mechanism remains elusive. In the present study, the chondrocytes were extracted from the cartilage tissues of New Zealand white rabbits, cultured in the presence of sodium nitroprusside（SNP）（2.5 mmol/L） and divided into five groups： 1, 5, 10, and 20 μmol/L SRT1720 groups and blank control group（0 μmol/L SRT1720）. MTT assay was used to detect the chondrocyte viability and proliferation, and DAPI staining and flow cytometry to measure the chondrocyte apoptosis. The expression levels of SIRT1, p53, NF-κB/p65, Bax, and peroxisome proliferator-activated receptor gamma coactivator 1-α（PGC-1α） were detected by Western blotting and the expression levels of SIRT1, type Ⅱ collagen, and aggrecan m RNA by RT-PCR. The results showed that in the SRT1720-treated groups, the nuclei of chondrocytes were morphologically intact and had uniform chromatin. In the blank control group, nuclear rupture into debris was observed in chondrocytes. With the SRT1720 concentration increasing, the chondrocyte viability increased, the apoptosis rate decreased, the protein expression levels of SIRT1 and PGC-1α and the m RNA expression levels of type Ⅱ collagen and aggrecan increased（P〈0.05）, and the expression levels of p53, NF-κB and bax decreased（P〈0.05）. It was suggested that SRT1720 inhibits chondrocyte apoptosis by activating the expression of SIRT1 via p53/bax and NF-κB/PGC-1α pathways.
基金
supported by grants from the National Natural Science Foundation of China(No.81272032)
Key Project of Shenzhen Science and Technology Plan(No.201101001)
