External factors
| External factors: |
Apigenin |
| Category: |
Chemical compounds |
| Experiment: |
SA-β-gal activity assay//Cell viability assay//Western blot |
| Description: |
Senescent cells were stained blue due to SA-β-gal activity, while only ~46%, and 23% of cells displayed SA-β-gal activity after pre-treatment with 10 and 20 μM apigenin, respectively.Apigenin pre-treatment also ameliorated H2O2-induced growth arrest and did not affect the normal growth speed of cells.Apigenin pre-treatment downregulated protein levels of ac-p53, p21Waf1/Cip1, and p16Ink4a and upregulated protein levels of p-Rb and cyclin D1.Interestingly, apigenin pre-treatment also significantly reduced cytosolic positive stain (blue) and abrogated DOXO-induced cell enlargement and flattened morphology.The percentage of SA-β-gal activity were ~48% and 19% at 10 and 20 μM apigenin treatment, respectively. Meanwhile, the decreases in ac-p53, p21Cip1/WAF1, and p16Ink4a and the increases in p-Rb and cyclin D1 were observed. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation//Downregulation |
| Official symbol(s): |
SIRT1//CD38 |
| Target gene experiment: |
RT-qPCR//Western blot |
| Target gene description: |
Apigenin induced expression of the SIRT1 protein and mRNA in a concentration- and time-dependent manner. Subsequently, SIRT1 activity was examined and were found to increase with apigenin concentration and time. Apigenin increased NAD+ and NAD+/NADH activity in a concentrationdependent manner, while it attenuated CD38 activity. Next, we examined whether the apigenin-induced reduction in CD38 activation altered levels of SIRT1. As shown in Figs. 5D and 5E, apigenin and CD38 siRNA inhibited CD38 expression and modulated SIRT1 levels. |
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| Regulatory pathway:
|
p53-p21Cip1/WAF1//p16Ink4a-Rb |
| R-EF-Pathway:
|
Downregulation |
| Pathway experiment: |
Western blot |
| Pathway description: |
Apigenin pre-treatment downregulated protein levels of ac-p53, p21Waf1/Cip1, and p16Ink4a and upregulated protein levels of p-Rb and cyclin D1.SIRT1 siRNA treatment decreased SIRT1 levels and the modulatory role of apigenin on ac-p53, p21Waf1/Cip1, p16Ink4a, p-Rb, and cyclin D1 was abolished. |
Aging network
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Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.
Pathway view
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