科研之友
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摘要
Context: Ternstroemia gymnanthera Sprague (Theaceae) possesses various known pharmacological properties. However, its anti-inflammatory activity has not been reported. Objective: The anti-inflammatory activity of Ternstroemia gymnanthera stem bark aqueous extract (TGSBE) was evaluated using LPS-stimulated RAW264.7 macrophages. Materials and methods: Cytotoxicity was assessed by MTT assay after 24 h with TGSBE (25-200 mu g/mL). Further testing used TGSBE at 100 and 200 mu g/mL. Griess and ELISA methods after 24 h with TGSBE determined NO and cytokine levels, respectively; then, mRNA levels (iNOS & cytokines) were analyzed by Quantitative-PCR after 12 h. NF-kappa B and MAPK were assessed by immunoblotting after TGSBE treatment for 12 h, followed by LPS for 30 min. Immunofluorescence assay was also performed for NF-kappa B. ROS and MMP, after 12 h with TGSBE, were determined by flow cytometry. The antioxidant potential of TGSBE was analyzed by ABTS assay. The Folin-Ciocalteu method determined the total phenolic content of TGSBE. LPS concentration was 0.5 mu g/mL. Results: TGSBE at 200 mu g/mL showed about 96.2% viability while suppressing the production of NO (88.99%), TNF alpha (24.38%), IL-6 (61.70%) and IL-1 beta (55.12%) and gene expression by 67.88, 45.24, 65.84, and 70.48%, respectively. TGSBE decreased ROS (79.26%) and improved MMP (48.01%); it inhibited translocation of NF-kappa B and MAPK activation. Radical scavenging activity was 50% at 402.17 mu g/mL (ascorbic acid standard: 88.8 mu g/mL). Total phenolic content was 240.9mg GAE/g. Discussion and conclusion: TGSBE suppresses the inflammatory response by inhibiting the NF-kappa B and MAPK cascades exhibiting therapeutic potential to treat inflammatory diseases associated with increased activation of macrophages.
- 出版日期2017
