A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation of GP
2.3. Total Flavonoid Content of GP
2.4. Cell Culture
2.5. NO Production Measurement
2.6. Assay for Intracellular Calcium Release
2.7. Multiplex Cytokine Assay
2.8. Assay for mRNA Expressions of Inflammatory Genes
2.9. Assay for Phosphorylation of P38 MAPK
2.10. Statistics
3. Results
3.1. Determination of Total Flavonoid Content of GP
3.2. Effects of GP on Levels of NO and Ca2+
3.3. Effect of GP on Cytokine Production
- -
- MIP-1α productions from RAW 264.7 with GP at concentrations of 25, 50, 100, and 200 µg/mL were 98.84 ± 0.7%, 97.78 ± 0.85%, 98.41 ± 0.52%, and 98.34 ± 0.66% of the control group treated with LPS only;
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- MIG were 69.02 ± 7.59%, 69.94 ± 6.38%, 79.75 ± 9.44%, and 76.38 ± 9.2%;
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- MCP-1 were 81.91 ± 3.86%, 91.69 ± 10.81%, 75.36 ± 7.39%, and 72.45 ± 1.89%;
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- GM-CSF were 92.83 ± 2.62%, 86.17 ± 4.82%, 76.12 ± 9.42%, and 90.23 ± 9.65%;
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- VEGF were 100.61 ± 10.56%, 83.55 ± 10.99%, 83.44 ± 8.75%, and 96.4 ± 4.14%;
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- IL-1β were 67.65 ± 10.6%, 93.43 ± 25.89%, 93.43 ± 15.8%, and 82.8 ± 6.69%;
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- LIX were 77.78 ± 5.39%, 85.26 ± 9.17%, 89.21 ± 6.45%, and 86.92 ± 9%;
- -
- LIF were 81.82 ± 9.14%, 84.81 ± 11.59%, 96.35 ± 1.02%, and 83.56 ± 3.29%;
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- IL-6 were 99 ± 0.4%, 98.31 ± 0.82%, 98.83 ± 0.27%, and 98.61 ± 0.59%.
3.4. Effect of GP on mRNA Expressions of Inflammatory Genes
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- Chop mRNA levels in LPS-activated RAW 264.7 with GP at concentrations of 25, 50, 100, and 200 µg/mL were 94.24 ± 4%, 81.17 ± 6.88%, 19.87 ± 9.02%, and 31.12 ± 4.13% of the control group treated with LPS only;
- -
- Camk2a were 98.74 ± 14.5, 28.17 ± 8.37%, 30.13 ± 10.88%, and 27.62 ± 0.56%;
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- Stat1 were 52.52 ± 6.34, 73.17 ± 8.11%, 57.45 ± 5.04%, and 55.89 ± 4.94%;
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- Stat3 were 51.04 ± 5.49, 68.85 ± 7.68%, 62.08 ± 5.93%, and 53.83 ± 4.5%;
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- Jak2 were 77.22 ± 10.46, 43.18 ± 5.75%, 44.82 ± 12.07%, and 50.42 ± 10.86%;
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- Fas were 71.59 ± 7.23, 49.15 ± 4.99%, 42.11 ± 3.66%, and 34.57 ± 4.67%;
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- Nos2 were 44.11 ± 5.77, 91.15 ± 9.71%, 67.8 ± 5.61%, and 78.73 ± 5.26%;
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- Ptgs2 were 49.65 ± 5.36, 90.5 ± 14.47%, 65.88 ± 3.94%, and 58.52 ± 6.52%.
3.5. Effect of GP on Phosphorylation of P38 MAPK
4. Discussion
5. Conclusions
Funding
Acknowledgments
Conflicts of Interest
References
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Name 1 | Forward Primer (5′–3′) | Reverse Primer (5′–3′) |
---|---|---|
Chop | CCACCACACCTGAAAGCAG | TCCTCATACCAGGCTTCCA |
Camk2a | AGCCATCCTCACCACTAT | ATTCCTTCACGCCATCATT |
Stat1 | TGAGATGTCCCGGATAGTGG | CGCCAGAGAGAAATTCGTGT |
Stat3 | GTCTGCAGAGT TCAAGCACCT | TCCTCAGTCACGATCAAGGAG |
Jak2 | TTGGTTTTGAATTATGGTGTCTGT | TCCAAATTTTACAAATTCTTGAACC |
Fas | CGCTGTTTTCCCTTGCTG | CCTTGAGTATGAACTCTTAACTGTGAG |
Nos2 | TGGAGGTTCTGGATGAGAGC | AATGTCCAGGAAGTAGGTGAGG |
Ptgs2 | TCAAACAGTTTCTCTACAACAACTCC | ACATTTCTTCCCCCAGCAA |
β-Actin | CTAAGGCCAACCGTGAAAAG | ACCAGAGGCATACAGGGACA |
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Park, W. A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway. Processes 2021, 9, 1632. https://doi.org/10.3390/pr9091632
Park W. A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway. Processes. 2021; 9(9):1632. https://doi.org/10.3390/pr9091632
Chicago/Turabian StylePark, Wansu. 2021. "A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway" Processes 9, no. 9: 1632. https://doi.org/10.3390/pr9091632