Cleome rutidosperma and Euphorbia thymifolia Suppress Inflammatory Response via Upregulation of Phase II Enzymes and Modulation of NF-κB and JNK Activation in LPS-Stimulated BV2 Microglia
Abstract
:1. Introduction
2. Results
2.1. Ethanol Extracts of C. rutidosperma (CR) and E. thymifolia (ET) Inhibited Nitric Oxide (NO) Production and Activation in Lipopolysaccharide (LPS)-Treated BV2 Cells
2.2. Ethanol Extracts of C. rutidosperma and E. thymifolia Down-Regulated Protein Expression of Inducible Nitric Oxide Synthase (iNOS) and Cyclooxygenase (COX)-2
2.3. Effects of Ethanol Extracts of C. rutidosperma and E. thymifolia on mRNA Expression of iNOS and COX-2
2.4. Effects of Ethanol Extracts of C. rutidosperma and E. thymifolia on Pro-inflammatory Cytokine Production and Expression
2.5. Ethanol Extracts of C. rutidosperma and E. thymifolia Induce Nrf2 Target Gene Expression
2.6. Effects of Ethanol Extracts of C. rutidosperma and E. thymifolia on LPS-Mediated Activation of MAPK and NF-κB
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Extraction
4.3. BV2 Cell Culture and Measurement of Nitrite, TNF and IL-6 Release
4.4. Cell Viability
4.5. Western Blotting Analysis
4.6. RNA Extraction and Reverse Transcription Real-Time PCR
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Antibody | Company | Catalog Number |
---|---|---|
α-tubulin | Sigma-Aldrich | T6199 |
NOS | Cell Signaling (Danvers, MA, USA) | 2977 |
COX-2 | Santa Cruz (Santa Cruz, CA, USA) | Sc-166475 |
HO-1 | Stressgen (San Diego, CA, USA) | SPA-895 |
Phospho-JNK1/2 (clone 81E11) | Cell Signaling | 4668 |
JNK2 (clone 56G8) | Cell Signaling | 9258 |
Phoshpo-p38 MAPK(clone 3D7) | Cell Signaling | 9215 |
p38 MAPK | Cell Signaling | 9212 |
Phospho-p65 NF-κB (clone 93H1) | Cell Signaling | 3033 |
p65 NF-κB | Cell Signaling | 3034 |
Gene | Primers (5′–3′) | Amplicon (bp) |
---|---|---|
β-actin | GGCTGTATTCCCCTCCATCG CCAGTTGGTAACAATGCCATGT | 154 |
iNOS | GTTCTCAGCCCAACAATACAAGA GTGGACGGGTCGATGTCAC | 127 |
COX-2 | TGAGCAACTATTCCAAACCAGC GCACGTAGTCTTCGATCACTATC | 74 |
IL-1β | TTCAGGCAGGCAGTATCACTC GAAGGTCCACGGGAAAGACAC | 75 |
CCL2 | TTAAAAACCTGGATCGGAACCAA GCATTAGCTTCAGATTTACGGGT | 121 |
HO-1 | AAGCCGAGAATGCTGAGTTCA GCCGTGTAGATATGGTACAAGGA | 100 |
GCLM | AAGCCCAGGATTGGGTGCCG GGTCGGTGAGCTGTGGGTGT | 141 |
NQO1 | AGGATGGGAGGTACTCGAATC TGCTAGAGATGACTCGGAAGG | 127 |
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Ding, H.-Y.; Wu, P.-S.; Wu, M.-J. Cleome rutidosperma and Euphorbia thymifolia Suppress Inflammatory Response via Upregulation of Phase II Enzymes and Modulation of NF-κB and JNK Activation in LPS-Stimulated BV2 Microglia. Int. J. Mol. Sci. 2016, 17, 1420. https://doi.org/10.3390/ijms17091420
Ding H-Y, Wu P-S, Wu M-J. Cleome rutidosperma and Euphorbia thymifolia Suppress Inflammatory Response via Upregulation of Phase II Enzymes and Modulation of NF-κB and JNK Activation in LPS-Stimulated BV2 Microglia. International Journal of Molecular Sciences. 2016; 17(9):1420. https://doi.org/10.3390/ijms17091420
Chicago/Turabian StyleDing, Hsiou-Yu, Pei-Shan Wu, and Ming-Jiuan Wu. 2016. "Cleome rutidosperma and Euphorbia thymifolia Suppress Inflammatory Response via Upregulation of Phase II Enzymes and Modulation of NF-κB and JNK Activation in LPS-Stimulated BV2 Microglia" International Journal of Molecular Sciences 17, no. 9: 1420. https://doi.org/10.3390/ijms17091420