| Description: |
It was determined that the expression level SA-β-gal in FLS increased with H/R intervention compared to the control group. Furthermore, in the H/R + TNF-α group, the expression level of SA-β-gal was further increased compared with the control group, H/R group and TNF-α group, which indicated a larger proportion of senescent cells. IL-1β and IL-6 levels in FLS were assessed using ELISA, and HMGB1, Casp8, p16, p21, MMP-3 and MMP-13 proteins were detected by western blotting. In addition, RT-qPCR was used to detect the mRNA expression levels of HMGB1, Casp8, p16, p21, MMP-3, MMP-13, IL-1β and IL-6. The results indicated that the protein and mRNA expression levels of HMGB1, Casp8, p16, p21, MMP-3, MMP-13, IL-1β and IL-6 were significantly increased in the H/R environment compared with the control. Moreover, the expression levels of all these factors were further significantly increased after pre-incubation with TNF-α before 25 h of H/R compared with the control group, H/R group and TNF-α group. |