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 (PGE 2 ) 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, PGE 2 , 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.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.