Anti‑inflammatory activity of 3,5,6,7,3',4'‑hexamethoxyflavone via repression of the NF‑κB and MAPK signaling pathways in LPS‑stimulated RAW264.7 cells

Son, Eun Suk; Park, Jeong‐Woong; Kim, Se‐Hee; Park, Hye Ran; Han, Woorijarang; Kwon, O-Chul; Nam, Jae Do; Jeong, Sung Hwan; Lee, Chang‐Soo

doi:10.3892/mmr.2020.11252

Cited by 12 publications

(2 citation statements)

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“…Inflammation is a natural host defense reaction process, which is divided into acute and chronic inflammation according to the duration ( 29 ). The main symptoms of acute inflammation include redness, swelling and pain ( 30 , 31 ).…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2021

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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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2021

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“…Oxypeucedanin hydrate alleviates rheumatoid arthritis various activities and external stimuli, including inflammation, differentiation, apoptosis, and gene expression [27,28]. MAPKtargeting inhibitors are synthesized for treating inflammatory diseases [29].…”

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confidence: 99%

Oxypeucedanin hydrate alleviates rheumatoid arthritis by inhibiting the TLR4-MD2/NF-κB/MAPK signaling axis

Liu,

Huo,

et al. 2024

ABBS

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Rheumatoid arthritis (RA) is an idiopathic and chronic autoimmune disease for which there are currently no effective treatments. Oxypeucedanin hydrate (OXH) is a natural coumarin known for its potent anti-inflammatory properties. However, further investigations are needed to determine its therapeutic efficacy in treating RA. In this study, we evaluate the anti-inflammatory activity of OXH by treating LPS-induced RAW264.7 macrophages. Our results show that OXH treatment reverses the changes in iNOS, COX-2, IL-1β, IL-6, and TNF-α levels. Additionally, OXH reduces ROS production. Further analysis reveals that OXH suppresses the activation of the NF-κB/MAPK pathway. CETSA results show that OXH competes with LPS for binding to the TLR4/MD2 complex. MST experiments demonstrate the specific affinity of OXH for the TLR4/MD2 complex, with a Kd value of 33.7 μM. Molecular docking analysis suggests that OXH binds to the pocket of the TLR4/MD2 complex and interacts with specific amino acids, such as GLY-343, LYS-388, and PHE-345. Molecular dynamics simulations further confirm this conclusion. Finally, we investigate the potential of OXH in treating RA using a collagen-induced arthritis (CIA) model in rats. OXH effectively ameliorates the symptoms of CIA, including improving body weight, reducing swelling and redness, increasing talus volume, and decreasing bone erosion. OXH also decreases the mRNA levels of pro-inflammatory factors in synovial tissue. Transcriptome enrichment analysis and western blot analysis confirm that OXH suppresses the NF-κB/MAPK pathway, which is consistent with our in vitro findings.

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L-theanine attenuates LPS-induced motor deficit in experimental rat model of Parkinson’s disease: emphasis on mitochondrial activity, neuroinflammation, and neurotransmitters

et al. 2023

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Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopamine neurons in the substantia nigra. The pathogenesis of PD, including oxidative stress, mitochondrial dysfunction, neuroin ammation, and neurotransmitter dysregulation. L-theanine is an amino acid found in green tea and has antioxidant, anti-in ammatory, and neuroprotective effects with a high BBB permeability.Therefore, the current study was designed to investigate the possible neuroprotective effect of L-theanine in lipopolysaccharide (LPS) induced motor de cits and striatal neurotoxicity in a rat model of PD. LPS was infused at a dose of 5 µg/5 µl PBS, stereotaxically into SNpc of rats. Treatment with L-theanine at (50 and 100 mg/kg; po), and Sinemet (36 mg/kg; po) was given from day 7 to 21 in of LPS injected rat.On a weekly basis all behavioral parameters were assessed, and animals were sacri ced on day 22. The striatum tissue of brain was isolated for biochemicals (Nitrite, GSH, catalase, SOD, mitochondrial complexes I and IV), neuroin ammatory markers (IL-1β, TNF-α, and IL-6), and neurotransmitters (serotonin, dopamine, norepinephrine, GABA, and glutamate) estimations. Results revealed that Ltheanine dose-dependently and signi cantly reversed motor de cits, assessed through locomotor and rotarod activity. Moreover, L-theanine attenuated biochemical markers, reduced oxidative stress, and neurotransmitters dysbalance in the brain. L-theanine treatment at 100 mg/kg; po substantially reduced these pathogenic events by increasing mitochondrial activity, restoring neurotransmitter levels, and inhibiting neuroin ammation. Moreover, data suggest that the positive effects of L-theanine on motor coordination may be mediated by the suppression of nuclear factor-κB (NF-κB) induced by LPS.

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Anti‑inflammatory activity of 3,5,6,7,3',4'‑hexamethoxyflavone via repression of the NF‑κB and MAPK signaling pathways in LPS‑stimulated RAW264.7 cells

Cited by 12 publications

References 45 publications

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

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

Oxypeucedanin hydrate alleviates rheumatoid arthritis by inhibiting the TLR4-MD2/NF-κB/MAPK signaling axis

L-theanine attenuates LPS-induced motor deficit in experimental rat model of Parkinson’s disease: emphasis on mitochondrial activity, neuroinflammation, and neurotransmitters

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Anti‑inflammatory activity of 3,5,6,7,3',4'‑hexamethoxyflavone via repression of the NF‑κB and MAPK signaling pathways in LPS‑stimulated RAW264.7 cells

Cited by 12 publications

References 45 publications

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

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

Oxypeucedanin hydrate alleviates rheumatoid arthritis by inhibiting the TLR4-MD2/NF-&kappa;B/MAPK signaling axis

L-theanine attenuates LPS-induced motor deficit in experimental rat model of Parkinson’s disease: emphasis on mitochondrial activity, neuroinflammation, and neurotransmitters

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Oxypeucedanin hydrate alleviates rheumatoid arthritis by inhibiting the TLR4-MD2/NF-κB/MAPK signaling axis