| External factors: | t-BHP |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Intervertebral disk |
| Experiment: | SA-β-gal activity assay |
| Description: | We conducted SA-β-Gal staining to label senescent NPSCs. The results showed that treatment with t-BHP resulted in a significant increase in the percentage and the staining intensity of SA-β-Gal-positive NPSCs in a dose-dependent manner. However, HSP70 activation significantly decreased the SA-β-Gal-positive rate, indicating that HSP70 impeded the process of t-BHP-induced senescence. |
| Target gene: | HSP70 |
| R-EF-Target gene: | Upregulation//Upregulation |
| Official symbol(s): | HSP70 |
| Target gene experiment: | Western blot |
| Target gene description: | 12 h treatment of t-BHP significantly decreased NPSC viability in a dose-dependent manner in all groups except 1 μM t-BHP-treated group com- pared with the control group. In addition, time-dependent cell viability loss was also observed after treating with 120 μM t-BHP for 3 h, 6 h, 9 h, 12 h, 18 h, or 24 h compared with the control group. Furthermore, we performed western blot analysis to investigate the promoting effects of t-BHP on HSP70 expres- sion in NPSCs. Our data indicated that t-BHP significantly stimulated the expression of HSP70 in human NPSCs in all doses except 200 μM group compared with the control group. In addition, treatment with t-BHP for 3 h, 6 h, 12 h, 18 h, or 24 h increased the expression of HSP70 compared with the control group. However, the expression of HSP70 partially declined after treatment with t-BHP for 24 h, indicating a HSP70 fluctuation in a time-dependent manner. |
| Regulatory pathway: | p53-p21//JNK-C-Jun |
| R-EF-Pathway: | Activation//Activation |
| Official symbol(s): | TP53-CDKN1A// |
| Pathway experiment: | Western blot//Western blot |
| Pathway description: | T-BHP treatment elevated the expression levels of p53 and p21. The protein expression of the p53/p21 pathway was determined by western blot analysis. T-BHP treatment elevated the expression levels of p53 and p21 compared with the control group. We also found that the activation of HSP70 significantly decreased the protein levels of p53 and p21.//T-BHP treatment elevated the levels of p-JNK/JNK ratio and p–c-Jun/c-Jun ratio in a dose-dependent manner. |
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