External factors
| External factors: |
Paeonol |
| Category: |
Chemical compounds |
| Experiment: |
MTT assay//SA-β-gal activity assay |
| Description: |
Paeonol (60–120 lM) notably increased the MRC-5 cell viability induced by 60 lM H2O2. So effective concentrations of paeonol (60, 90, 120 lM) were used in subsequent experiments. Moreover, aging MRC-5 cells was evaluated by senescence marker SA-β-gal, and paeonol treatment weakened 60 lM H2O2-induced senescence of MRC-5 cells, as indicated by reduction of the SA-β-gal activity. After the aging MRC-5 cells were treated with paeonol (60, 90, 120 lM) for 24 h, the results showed that paeonol remarkably reduced the number of blue-stained cells in a dosedependent manner. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation |
| Target gene experiment: |
Western blot |
| Target gene description: |
To further study the suppression of ROS level and cellular senescence in H2O2- exposed MRC-5 cells by paeonol, we isolated the nuclear and cytosolic fractions to detect Nrf2 protein levels, and Lamin B was used as the loading control of nuclear Nrf2. The results showed that nuclear Nrf2 protein was increased under H2O2 exposure, but was significantly upregulated by paeonol treatment. Interestingly, cytosolic Nrf2 protein was decreased under oxidative stress, but alteration in the expression of cytosolic Nrf2 appeared to be similar to that of nuclear Nrf2 when treated with paeonol. |
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| Regulatory pathway:
|
Id-1//p16//p53-p21 |
| R-EF-Pathway:
|
Upregulation//Downregulation//Downregulation |
| Official symbol(s): |
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ID1////TP53-CDKN1A
|
| Pathway experiment: |
SA-β-gal activity assay//Western blot |
| Pathway description: |
So we have decided to ascertain whether paeonol could disrupt the expression of Id-1, p16INK4a, and p53/ p21Waf1/Cip1 pathway to weaken senescence in MRC-5 cells.The expression of Id-1 in aging MRC-5 cells treated with paeonol (60, 90, 120 lM) for 24 h was gradually evaluated compared to H2O2 group; meanwhile, paeonol decreased p16INK4a, p53,and p21Waf1/Cip1 (downstream protein of p53) expression in the aging MRC-5 cells.This effect has been extensively studied that Id-1 delayed cellular senescence in human fibroblasts through suppression of p16INK4a, a known mediator of cellular senescence. Elevation of p53 and p21Waf1/Cip1 levels has been observed in aging fibroblasts, which would lead to a promotion in senescence.Our results showed that paeonol attenuated aging MRC-5 cells in part via regulation of Id-1, p16INK4a, and p53/p21Waf1/Cip1 pathways. |
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.
Pathway view
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