Anti-Inflammatory Activity and ROS Regulation Effect of Sinapaldehyde in LPS-Stimulated RAW 264.7 Macrophages

Abstract

We evaluated the anti-inflammatory effects of SNAH in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by performing nitric oxide (NO) assays, cytokine enzyme-linked immunosorbent assays, Western blotting, and real-time reverse transcription-polymerase chain reaction analysis. SNAH inhibited the production of NO (nitric oxide), reactive oxygen species (ROS), tumor necrosis factor (TNF)-α, and interleukin (IL)-6. Additionally, 100 μM SNAH significantly inhibited total NO and ROS inhibitory activity by 93% (p < 0.001) and 34% (p < 0.05), respectively. Protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) stimulated by LPS were also decreased by SNAH. Moreover, SNAH significantly (p < 0.001) downregulated the TNF-α, IL-6, and iNOS mRNA expression upon LPS stimulation. In addition, 3–100 µM SNAH was not cytotoxic. Docking simulations and enzyme inhibitory assays with COX-2 revealed binding scores of −6.4 kcal/mol (IC50 = 47.8 μM) with SNAH compared to −11.1 kcal/mol (IC50 = 0.45 μM) with celecoxib, a known selective COX-2 inhibitor. Our results demonstrate that SNAH exerts anti-inflammatory effects via suppression of ROS and NO by COX-2 inhibition. Thus, SNAH may be useful as a pharmacological agent for treating inflammation-related diseases.

Poly ε-Caprolactone Nanoparticles for Sustained Intra-Articular Immune Modulation in Adjuvant-Induced Arthritis Rodent Model

“…The effect of Lfd-NPs on the expression of tumor necrosis factor alpha (TNF-α) and Interleukin (IL)-6 in the supernatant of LPS-treated THP-1 human monocytes and RAW 264.7 murine macrophage cells was investigated via an enzyme-linked immunosorbent assay (ELISA) ( Figure 6 ). TNF-α has been linked to a variety of signaling processes inside cells that result in necrosis or apoptosis [ 58 ], while IL-6 is known to contribute to an acute phase immune response via inducing the synthesis of acute phase proteins such as CRP [ 59 ]. As shown in Figure 6 , in cultures of LPS-treated cells, both TNF-α and IL-6 concentrations were significantly increased, compared to untreated control cells ( Figure 6 A–D).…”

Anti-inflammatory effect of a triterpenoid from Balanophora laxiflora: results of bioactivity-guided isolation

“…ROS signaling is also essential for the activation of inflammasomes, molecular platforms playing a key role in the processing of pro-inflammatory cytokines, such as IL-1β, IL-18, and IL-33 ( Harijith et al., 2014 ; Pham and Park 2020 ). Unsurprisingly, ROS inhibitors may ameliorate the inflammatory conditions triggered by diverse stimuli and prevent the development and progression of inflammation-linked complications ( Yang et al., 2018 ; Baek et al., 2020 ). In line with previous reports, we observed that blockade of ROS signaling by either suppressing NOX-mediated ROS production with DPI or scavenging ROS using NAC leads to decreased COX-2 expression in macrophages, confirming the involvement of ROS in modulating COX-2 in our experimental models.…”

Characterization, Antioxidant and Anti-Inflammation Capacities of Fermented Flammulina velutipes Polyphenols

“…The NO contents and ROS levels in the supernatants were determined after centrifugation at 1000 rpm for 5 min and incubation for 24 h by measuring the fluorescence intensity at 485 nm and 540 nm, according to the instructions. Then, the NO contents and ROS levels were calculated according to standard curve and formula (1), respectively [ 49 ]. The cell fluorescence images were analyzed using a Nikon fluorescence microscope (ECLIPSE NI, Tokyo, Japan).…”