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10.3390/life13020595
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Article

Helixor-M Suppresses Immunostimulatory Activity through TLR4-Dependent NF-κB Pathway in RAW 264.7 Cells

by
Doil Park
1,2,
Hyun Min Ko
1,2,
Wona Jee
1,2,
So Mi Park
1,2,
Ye Rin Park
1,2,
Ji Hoon Jung
1,
Hyung Suk Kim
1,3,
Won Seok Chung
1,3,
Sang Ki Kim
4,
Jong Sup Chung
4 and
Hyeung Jin Jang
1,*
1
College of Korean Medicine, Kyung Hee University, 24 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea
2
Department of Science in Korean Medicine, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea
3
Department of Korean Rehabilitation Medicine, Kyung Hee University Medical Center, Seoul 02447, Republic of Korea
4
Dalim Biotech, 33, Sinpyeong-ro, Jijeong-myeon, Wonju-si 26348, Republic of Korea
*
Author to whom correspondence should be addressed.
Life 2023, 13(2), 595; https://doi.org/10.3390/life13020595
Submission received: 26 January 2023 / Revised: 10 February 2023 / Accepted: 17 February 2023 / Published: 20 February 2023
(This article belongs to the Special Issue Macrophages in Human Diseases: From Pathophysiology to Therapeutic Targets)
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Abstract

Inflammation causes a protective immune response, which can be observed by examining the inflammatory responses of macrophages. Macrophages release various immunostimulatory factors when destroying external pathogens. We induced lipopolysaccharides (LPS) in RAW 264.7 cells, a macrophage cell line, to determine whether Helixor-M can cause immuno-suppression. Helixor-M is known to have anticancer and immune effects. However, an indicator that regulates immunity has not been clearly confirmed. To this end, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted to confirm Helixor-M was not cytotoxic. Western blotting and real-time polymerase chain reaction (RT-PCR) confirmed the anti-inflammatory effects. Additionally, immunofluorescence assay confirmed the translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65, a representative inflammatory pathway. Helixor-M was found to be non-cytotoxic, induce the NF-κB pathway, and reduce the levels of pro-inflammatory cytokine and mitogen-activated protein kinase (MAPK). We found Helixor-M affected the PI3K/AKT/JNK pathway. Therefore, we confirmed Helixor-M acts as an anti-inflammatory agent through NF-κB, TLR4 and PI3K inhibition and that it could be an effective immunosuppressive drug.
Keywords: Helixor M; NF-κ B; RAW 264.7 cells; immune; MAPK; AKT; PI3K Helixor M; NF-κ B; RAW 264.7 cells; immune; MAPK; AKT; PI3K

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MDPI and ACS Style

Park, D.; Ko, H.M.; Jee, W.; Park, S.M.; Park, Y.R.; Jung, J.H.; Kim, H.S.; Chung, W.S.; Kim, S.K.; Chung, J.S.; et al. Helixor-M Suppresses Immunostimulatory Activity through TLR4-Dependent NF-κB Pathway in RAW 264.7 Cells. Life 2023, 13, 595. https://doi.org/10.3390/life13020595

AMA Style

Park D, Ko HM, Jee W, Park SM, Park YR, Jung JH, Kim HS, Chung WS, Kim SK, Chung JS, et al. Helixor-M Suppresses Immunostimulatory Activity through TLR4-Dependent NF-κB Pathway in RAW 264.7 Cells. Life. 2023; 13(2):595. https://doi.org/10.3390/life13020595

Chicago/Turabian Style

Park, Doil, Hyun Min Ko, Wona Jee, So Mi Park, Ye Rin Park, Ji Hoon Jung, Hyung Suk Kim, Won Seok Chung, Sang Ki Kim, Jong Sup Chung, and et al. 2023. "Helixor-M Suppresses Immunostimulatory Activity through TLR4-Dependent NF-κB Pathway in RAW 264.7 Cells" Life 13, no. 2: 595. https://doi.org/10.3390/life13020595

APA Style

Park, D., Ko, H. M., Jee, W., Park, S. M., Park, Y. R., Jung, J. H., Kim, H. S., Chung, W. S., Kim, S. K., Chung, J. S., & Jang, H. J. (2023). Helixor-M Suppresses Immunostimulatory Activity through TLR4-Dependent NF-κB Pathway in RAW 264.7 Cells. Life, 13(2), 595. https://doi.org/10.3390/life13020595

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