Cudratricusxanthone O Inhibits H2O2-Induced Cell Damage by Activating Nrf2/HO-1 Pathway in Human Chondrocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Plant Materials and Isolation of Compounds
2.3. Cell Culture
2.4. Cell Viability and Coefficient Assays
2.5. Western Blot Analysis
2.6. Cytosolic and Nuclear Protein Extraction
2.7. Measurement of ROS Generation
2.8. RT-qPCR Analysis
2.9. Annexin V-FITC/PI Apoptosis Assay
2.10. Statistical Analysis
3. Results
3.1. CTO Is Not Cytotoxic to SW1353 Chondrocytes
3.2. CTO Induces HO-1 Expression by Promotion of Nrf2 Translocation
3.3. CTO Suppresses ROS Production by Inducing SOD and CAT in SW1353 Cells
3.4. CTO Up-Regulates Chondrocytes-Specific Genes but Inhibits the Expression of MMPs in H2O2 Treated SW1353 Cells
3.5. CTO Inhibits Apoptotic Pathway Induced by Treatment with H2O2 in SW1353 Cells
3.6. Upregulated HO-1 by Pre-Treatment with CTO Protects SW1353 Cells from ROS Production Induced by H2O2
3.7. Induced HO-1 by CTO Regulates Chondrocytes-Specific Genes and Expression of mmps in H2O2 Treated SW1353 Cell
3.8. Enhanced HO-1 by CTO Shows Protective Effect from Apoptosis Induced by H2O2 in SW1353 Cells
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Target Gene | Sequence | |
---|---|---|
col2a1 | Forward (5′-3′) | TGGACGCCATGAAGGTTTTCT |
Reverse (3′-5′) | TGGGAGCCAGATTGTCATCTC | |
aggrecan | Forward (5′-3′) | GAAGTGGCGTCCAAACCAA |
Reverse (3′-5′) | CGTTCCATTCACCCCTCTCA | |
timp1 | Forward (5′-3′) | AATTCCGACCTCGTCATCAG |
Reverse (3′-5′) | GTTGTGGGACCTGTGGAAGT | |
mmp3 | Forward (5′-3′) | GGT GTG GAG TTC CTG ATG TTG |
Reverse (3′-5′) | AGC CTG GAG AAT GTG AGT GG | |
mmp13 | Forward (5′-3′) | TCA GGA AAC CAG GTC TGG AG |
Reverse (3′-5′) | TGA CGC GAA CAA TAC GGT TA | |
adamts | Forward (5′-3′) | TTCCACGGCAGTGGTCTAAAG |
Reverse (3′-5′) | CCACCAGGCTAACTGAATTACG | |
sod | Forward (5′-3′) | GGGAGATGGCCCAACTACTG |
Reverse (3′-5′) | CCAGTTGACATCGAACCGTT | |
cat | Forward (5′-3′) | ATGGTCCATGCTCTCAAACC |
Reverse (3′-5′) | CAGGTCATCCAATAGGAAGG | |
bcl2 | Forward (5′-3′) | AGG CTG GGA TGC CTT TGT GG |
Reverse (3′-5′) | GGG CAG GCA TGT TGA CTT CAC | |
bax | Forward (5′-3′) | TTCTGACGGCAACTTCAACTGG |
Reverse (3′-5′) | AGGAAGTCCAATGTCCAGCC | |
caspase3 | Forward (5′-3′) | GGAATTGATGCGTGATGTT |
Reverse (3′-5′) | TGGCTCAGAAGCACACAAAC | |
gapdh | Forward (5′-3′) | ACCCAGAAGACTGTGGATGG |
Reverse (3′-5′) | CACATTGGGGGTAGGAACAC |
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Kim, E.-N.; Lee, H.-S.; Jeong, G.-S. Cudratricusxanthone O Inhibits H2O2-Induced Cell Damage by Activating Nrf2/HO-1 Pathway in Human Chondrocytes. Antioxidants 2020, 9, 788. https://doi.org/10.3390/antiox9090788
Kim E-N, Lee H-S, Jeong G-S. Cudratricusxanthone O Inhibits H2O2-Induced Cell Damage by Activating Nrf2/HO-1 Pathway in Human Chondrocytes. Antioxidants. 2020; 9(9):788. https://doi.org/10.3390/antiox9090788
Chicago/Turabian StyleKim, Eun-Nam, Hyun-Su Lee, and Gil-Saeng Jeong. 2020. "Cudratricusxanthone O Inhibits H2O2-Induced Cell Damage by Activating Nrf2/HO-1 Pathway in Human Chondrocytes" Antioxidants 9, no. 9: 788. https://doi.org/10.3390/antiox9090788