External factors
| External factors: |
Hypoxia |
| Phenotype: |
Osteoarthritis |
| Tissue type: |
Synovial tissue |
| Experiment: |
SA-β-gal activity assay//Immunofluorescence//Western blot//ELISA//qRT-PCR |
| 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 24 h of H/R compared with the control group, H/R group and TNF-α group. |
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Regulatory relationship
| Official symbol(s): |
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MAPK8
|
| Pathway experiment: |
qRT-PCR//Western blot |
| Pathway description: |
Western blotting was used to assess the protein expression levels of JNK, p53, Bcl-2 and Bax. In addition, the mRNA expression levels of JNK, p53, Bax and Bcl-2 were detected by RT-qPCR. It was demonstrated that, compared with the control group, H/R intervention decreased the expression level of Bcl-2, and enhanced the expression levels of JNK, p53 and Bax. Therefore, the present results indicated that H/R may be important in activating the JNK signaling pathway to promote senescence. |
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.