External factors
| External factors: |
Glucose Restriction |
| Experiment: |
SA-β-gal activity assay//Cell proliferation assay |
| Description: |
The lifespan of all three human fibroblasts cell lines grown in GR medium was extended by an additional 2–4 weeks accompanied by an additional 5–10 PDs, which constituted a 23–67 prolongation of lifespan depending on different types of cells in GR medium.We found a low proportion of senescent cells in all three types of fibroblasts at the early stage of cell proliferation both in NG and GR medium-treated cells. Further, no prominen differences in SA-β-gal activity were observed between fibroblast cells with the treatment of NG and GR medium at early passage. However, successive subculture resulted in significantly increased senescent cells accumulation in NG medium versus GR medium at late passage. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation |
| Official symbol(s): |
SIRT1 |
| Target gene experiment: |
Western blot//qRT-PCR |
| Target gene description: |
We discovered that GR significantly increased SIRT1 mRNA and protein levels expression, most notably in late passage of cell growth and that this applied to all of the cell lines we examined. |
| Regulatory pathway:
|
p16-Rb//Akt-p70S6K1 |
| R-EF-Pathway:
|
Downregulation//Activation |
| Official symbol(s): |
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CDKN2A-RB1//AKT
|
| Pathway experiment: |
Western blot |
| Pathway description: |
Our results revealed a gradually increased p16 expression was accompanied by reduced expression of phosphorylated Rb (Ser-795) in NG medium during the cellular passaging . However,in GR medium, p16 protein signals were barely detected and expression of phosphorylated Rb was increased accordingly.Phosphorylated Akt and Akt protein kinase showed an increased expression trend during increased PDs in response to GR.p70S6K1 is a downstream target of Ak,GR caused a decreased expression of mTOR and its dependent protein kinases, phosphorylated p70S6K1 (Thr-389, Thr-421/Ser-424), whereas it did not affect the expression of p70S6K1 total protein, compared with the protein expression in NG. |
Aging network
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.