Open Access

α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway

  • Authors:
    • Jiang Tao Zhou
    • Kai Da Ren
    • Jing Hou
    • Jie Chen
    • Guan'e Yang
  • View Affiliations

  • Published online on: September 14, 2021     https://doi.org/10.3892/mmr.2021.12439
  • Article Number: 799
  • Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

α‑rhamnrtin‑3‑α‑rhamnoside (ARR) is the principal compound extracted from Loranthus tanakae Franch. & Sav. However, its underlying pharmacological properties remain undetermined. Inflammation is a defense mechanism of the body; however, the excessive activation of the inflammatory response can result in physical injury. The present study aimed to investigate the effects of ARR on lipopolysaccharide (LPS)‑induced RAW264.7 macrophages and to determine the underlying molecular mechanism. A Cell Counting Kit‑8 assay was performed to assess cytotoxicity. Nitric oxide (NO) production was measured via a NO colorimetric kit. Levels of prostaglandin E2 (PGE2) and proinflammatory cytokines, IL‑1β and IL‑6, were detected using ELISAs. Reverse transcription‑quantitative (RT‑q)PCR analysis was performed to detect the mRNA expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase‑2 (COX‑2), IL‑6 and IL‑1β in LPS‑induced RAW246.7 cells. Western blotting, immunofluorescence and immunohistochemistry analyses were performed to measure the expression levels of NF‑κB and nuclear factor‑erythroid 2‑related factor 2 (Nrf2) signaling pathway‑related proteins to elucidate the molecular mechanisms of the inflammatory response. The results of the cytotoxicity assay revealed that doses of ARR ≤200 µg/ml exhibited no significant effect on the viability of RAW264.7 cells. The results of the Griess assay demonstrated that ARR inhibited the production of NO. In addition, the results of the ELISAs and RT‑qPCR analysis discovered that ARR reduced the production of the proinflammatory cytokines, IL‑1β and IL‑6, as well as the proinflammatory mediators, PGE2, iNOS and COX‑2, in LPS‑induced RAW264.7 cells. Immunohistochemical analysis demonstrated that ARR inhibited LPS‑induced activation of TNF‑associated factor 6 (TRAF6) and NF‑κB p65 signaling molecules, while reversing the downregulation of the NOD‑like receptor family CARD domain containing 3 (NLRC3) signaling molecule, which was consistent with the results of the western blotting analysis. Immunofluorescence results indicated that ARR reduced the increase of NF‑κB p65 nuclear expression induced by LPS. Furthermore, the results of the western blotting experiments also revealed that ARR upregulated heme oxygenase‑1, NAD(P)H quinone dehydrogenase 1 and Nrf2 pathway molecules. In conclusion, the results of the present study suggested that ARR may exert anti‑inflammatory effects by downregulating NF‑κB and activating Nrf2‑mediated inflammatory responses, suggesting that ARR may be an attractive anti‑inflammatory candidate drug.
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November-2021
Volume 24 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Zhou JT, Ren KD, Hou J, Chen J and Yang G: α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway. Mol Med Rep 24: 799, 2021
APA
Zhou, J.T., Ren, K.D., Hou, J., Chen, J., & Yang, G. (2021). α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway. Molecular Medicine Reports, 24, 799. https://doi.org/10.3892/mmr.2021.12439
MLA
Zhou, J. T., Ren, K. D., Hou, J., Chen, J., Yang, G."α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway". Molecular Medicine Reports 24.5 (2021): 799.
Chicago
Zhou, J. T., Ren, K. D., Hou, J., Chen, J., Yang, G."α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway". Molecular Medicine Reports 24, no. 5 (2021): 799. https://doi.org/10.3892/mmr.2021.12439