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Article

Novel and Stable Dual-Color IL-6 and IL-10 Reporters Derived from RAW 264.7 for Anti-Inflammation Screening of Natural Products

1
Biomedical Research Institute, National institute of Advance Industrial Science and Technology, Tsukuba, Ibaraki 305-8566, Japan
2
Health Research Institute, National institute of Advance Industrial Science and Technology, Takamatsu, Kagawa 761-0395, Japan
3
Plant Research International, Wageningen University and Research Centre, 6708 PB Wageningen, The Netherlands
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(18), 4620; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms20184620
Received: 2 September 2019 / Revised: 13 September 2019 / Accepted: 16 September 2019 / Published: 18 September 2019
(This article belongs to the Special Issue Nutraceuticals in Cancer and Disease Prevention)
Our previous study suggested that the interleukin (IL)-6 and IL-10 could serve as good biomarkers for chronic inflammatory disease. We previously established an IL-6 and IL-10 reporters assay that could examine reporter activity along with the reference gene in LPS-induced RAW 264.7 cells. In this study, we described new and stable RAW 264.7 derived dual-color IL-6/gapdh and IL-10/gapdh reporters. This assay allowed us to easily determine relative IL-6 and IL-10 levels with 96-well plate within one step. We evaluated the relative IL-6 and IL-10 levels in the LPS-induced stable cells testing 52 natural products by real-time bioluminescence monitoring and time-point determination using a microplate luminometer. The relative IL-6 and IL-6/IL-10 values decreased by the crude ethanol extracts from nutmeg and by 1′S-1′-acetoxychavicol from greater galangal using real-time bioluminescence monitoring. At the same time, the relative IL-10 was induced. The relative IL-6 and IL-6/IL-10 decreased by crude ethanol extracts from nutmeg and 1′S-1′-acetoxychavicol acetate at 6 h. Only crude ethanol extract from nutmeg induced IL-10 at 6 h. We suggested that the use of these stable cells by real-time monitoring could serve as a screening assay for anti-inflammatory activity and may be used to discover new drugs against chronic inflammatory disease. View Full-Text
Keywords: IL-6; IL-10; luciferase; anti-inflammation IL-6; IL-10; luciferase; anti-inflammation
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MDPI and ACS Style

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://0-doi-org.brum.beds.ac.uk/10.3390/ijms20184620

AMA Style

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://0-doi-org.brum.beds.ac.uk/10.3390/ijms20184620

Chicago/Turabian Style

Saiki, 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://0-doi-org.brum.beds.ac.uk/10.3390/ijms20184620

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