Novel and Stable Dual-Color IL-6 and IL-10 Reporters Derived from RAW 264.7 for Anti-Inflammation Screening of Natural Products
Abstract
:1. Introduction
2. Results and Discussion
3. Materials and Methods
3.1. Cell Culture
3.2. Extraction, Isolation, and Purification of 1′S-1′-Acetoxychavicol Acetate
3.3. Analysis of Nuclear Magnetic Resonance Spectroscopy (NMR)
3.4. Nitric Oxide (NO) Production
3.5. Cell Proliferation Assay
3.6. Generation of Stable RAW 264.7 Cell Lines
3.7. Determination IL-6 and IL-10 Level by Stable Cells Using a Bioluminescence Measurement System for Living Cells by Real-Time Monitoring
3.8. Determination IL-6 and IL-10 Level by Stable Cells Using a Microplate Luminometer
3.9. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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NO Production | Relative IL-6/Gapdh | Relative IL-10/Gapdh | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
Extract Conc. (μg/mL) | Extract Conc. (μg/mL) | Extract Conc. (μg/mL) | ||||||||
No. | Sample Name | 150 | 100 | 50 | 75 | 50 | 25 | 75 | 50 | 25 |
1 | cinnamon | 0 | 0 | ● | ● | ● | ||||
2 | turmeric | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
3 | nutmeg | 0 | 0 | 0 | 0 | 0 | ● | ● | ● | |
4 | black pepper | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
5 | white pepper | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
6 | red pepper | ● | ● | ● | ||||||
7 | oregano | 0 | 0 | ● | ● | |||||
8 | basil | ● | ||||||||
9 | parsley | 0 | ● | ● | ● | |||||
10 | laurel | 0 | 0 | 0 | 0 | 0 | 0 | ● | ● | ● |
11 | cardamom | 0 | ● | |||||||
12 | cumin | 0 | ● | ● | ||||||
13 | cilantro | 0 | ● | ● | ● | |||||
14 | fennel | 0 | 0 | 0 | ● | ● | ● | |||
15 | anise seed | 0 | 0 | 0 | ● | ● | ● | |||
16 | allspice | ● | ● | ● | ||||||
17 | caraway | ● | ● | ● | ||||||
18 | clove | 0 | ||||||||
19 | Japanese pepper | 0 | 0 | 0 | ||||||
20 | ginger | 0 | ● | ● | ● | |||||
21 | star anise | ● | ||||||||
22 | celery seed | ● | ● | ● | ||||||
23 | long Pepper | 0 | 0 | 0 | 0 | 0 | ● | ● | ● | |
24 | fenogreek | ● | ● | ● | ||||||
25 | mace | 0 | 0 | 0 | ● | ● | ● | |||
26 | dil Seed | ● | ● | ● | ||||||
27 | kaffir lime leaf | 0 | 0 | 0 | ● | ● | ● | |||
28 | summer savory | ● | ● | |||||||
29 | kaba | 0 | 0 | ● | ● | ● | ||||
30 | sage | 0 | 0 | |||||||
31 | thyme | ● | ● | ● | ||||||
32 | taragon | 0 | ● | ● | ● | |||||
33 | marjoram | ● | ● | ● | ||||||
34 | rosemary | 0 | 0 | 0 | 0 | |||||
35 | Chinese pepper | 0 | 0 | 0 | 0 | |||||
36 | liquorice | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
37 | garlic | ● | ● | ● | ||||||
38 | green Pepper | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
39 | paprika | ● | ● | ● | ||||||
40 | horse radish | ● | ● | ● | ||||||
41 | mustard | ● | ● | ● | ||||||
42 | curry leaf | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
43 | spearmint | |||||||||
44 | dillweed | |||||||||
45 | peppermint | 0 | 0 | |||||||
46 | lemongrass | 0 | 0 | 0 | 0 | 0 | ||||
47 | Saffron | |||||||||
extract conc. (μg/mL) | 20 | 10 | 5 | |||||||
48 | greater galangal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ● |
extract conc. (μg/mL) | 150 | 100 | 50 | |||||||
49 | crude polysaccharide extract from Agaricus b. | ● | ● | ● | ||||||
50 | crude polyphenol extract. from Agaricus b | 0 | 0 | 0 | ● | ● | ● | |||
51 | crude polysaccharide extract from Ganoderma l. | ● | ● | ● | ||||||
52 | crude polysaccharide extract from Phellinus l. | 0 | 0 | 0 | 0 | 0 | ||||
0 | decreased | |||||||||
● | increased |
ACA | 1.3 | ACA | 1.3 | |
---|---|---|---|---|
A. Khalijah, et al. | H. Azuma et al. | |||
Carbon Number | 13C (ppm) | 13C (ppm) | 1H (ppm) | 1H (ppm) |
1 | 150.5, s | 150.6, s | ||
OCOCH3 | 169.4, s | 169.4, s | ||
OCOCH3 | 21.2, q | 21.2, q | 2.29 (s, 3H) | 2.25 (s, 3H) |
2 | 121.7, d | 121.8, d | 7.08 (d, 8.5, 1H) | 7.07 (d, 8.5, 1H) |
3 | 128.5, d | 128.6, d | 7.37 (d, 8.5, 1H) | 7.36 (d, 8.5, 1H) |
4 | 136.5, s | 136.6, s | ||
5 | 128.5, d | 128.6, d | 7.37 (d, 8.5, 1H) | 7.36 (d, 8.5, 1H) |
6 | 121.7, d | 121.8, d | 7.08 (d, 8.5 1H) | 7.07 (d, 8.5, 1H) |
1′ | 75.6, d | 75.6, d | 6.26 (d, 5.9, 1H) | 6.26 (d, 5.9, 1H) |
OCOCH3 | 169.7, s | 169.8, s | ||
OCOCH3 | 21.3, q | 21.3, q | 2.10 (s, 3H) | 2.10 (s, 3H) |
2′ | 136.1, d | 136.2, d | 5.98 (ddd, 17.1, 10.5, 5.9, 1H) | 5.98 (ddd, 17.1, 10.7, 6.1, 1H) |
3′ | 117.2, t | 117.2, t | 5.30 (d, 17.1, 1H), 5.25 (d, 10.5, 1H) | 5.29 (d, 17.1, 1H), 5.23 (d, 10.5, 1H) |
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Saiki, P.; Kawano, Y.; Nakajima, Y.; Van Griensven, L.J.L.D.; Miyazaki, K. Novel and Stable Dual-Color IL-6 and IL-10 Reporters Derived from RAW 264.7 for Anti-Inflammation Screening of Natural Products. Int. J. Mol. Sci. 2019, 20, 4620. https://doi.org/10.3390/ijms20184620
Saiki P, Kawano Y, Nakajima Y, Van Griensven LJLD, Miyazaki K. Novel and Stable Dual-Color IL-6 and IL-10 Reporters Derived from RAW 264.7 for Anti-Inflammation Screening of Natural Products. International Journal of Molecular Sciences. 2019; 20(18):4620. https://doi.org/10.3390/ijms20184620
Chicago/Turabian StyleSaiki, Papawee, Yasuhiro Kawano, Yoshihiro Nakajima, Leo J. L. D. Van Griensven, and Koyomi Miyazaki. 2019. "Novel and Stable Dual-Color IL-6 and IL-10 Reporters Derived from RAW 264.7 for Anti-Inflammation Screening of Natural Products" International Journal of Molecular Sciences 20, no. 18: 4620. https://doi.org/10.3390/ijms20184620