Silveira Leonardo Lopes, Dias Manoela Maciel Dos Santos, Pelinsari Silvânia Mól, de Paula Rosinéa Aparecida, Castro Adriano Simões Barbosa, Almeida Vera Lúcia de, Gonçalves Reggiani Vilela
Department of General Biology, Federal University of Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil.
Department of Animal Biology, Federal University of Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil.
J Ethnopharmacol. 2025 Jun 12;349:119973. doi: 10.1016/j.jep.2025.119973. Epub 2025 May 12.
Plants belonging to the Meliaceae family, such as Trichilia silvatica C. DC., known as catiguá-branco, have attracted considerable interest in phytochemical research due to their diverse and significant secondary metabolites. Trichilia silvatica has traditionally been employed in Brazilian medicine to treat inflammatory disorders. Moreover, studies have reported its antioxidant and antimicrobial properties, highlighting its potential therapeutic applications.
This study aimed to evaluate the potential of Trichilia silvatica leaf and stem extracts in modulating OxInflammation in RAW264.7 macrophage cells following exposure to lipopolysaccharide (LPS) or hydrogen peroxide (HO) and to elucidate the underlying mechanisms of action.
The phytochemical composition of the extracts was characterized using thin-layer chromatography (TLC), HPLC equipped with a reversed-phase Hypersil C-18 column, and spectrophotometric method. Their antioxidant activity was evaluated using the 2,2-difenil-1-picrilhidrazil (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assays. Cell viability was assessed via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alongside the determination of catalase (CAT) and superoxide dismutase (SOD) enzyme activities, as well as nitric oxide (NO) production in cells treated with the extracts and subsequently stimulated with HO. Gene expression levels of Factor nuclear kappa B (NF-κB), Ciclooxygenase 2 (COX-2), Tumor necrosis factor alpha (TNF-α), Interleukin 10 (IL-10), and Hypoxia-inducible factor-1 (HIF-1) were quantified using RT-qPCR.
Trichilia silvatica extracts revealed the presence of terpenes/steroids, coumarins, condensed tannins, and phenolic acids, including chlorogenic and caffeic acids. The findings indicate that the leaf extract at 100 μg/ml and the stem extract at 100 μg/ml and 250 μg/ml preserved or enhanced cell viability, conferring protection against HO-induced oxidative stress. These concentrations significantly increased CAT activity, whereas SOD activity remained unaffected. Nitric oxide production was significantly reduced when cells were treated with 100 μg/ml and 250 μg/ml of both leaf and stem extracts. Moreover, FRAP value revealed an increase in antioxidant capacity at 250 μg/mL. Both leaf and stem extracts, at 100 μg/mL and 250 μg/mL, exhibited a DPPH radical scavenging capacity exceeding 75 % and downregulated the expression of pro-inflammatory cytokines, including NF-κB, TNF-α, and COX-2. Notably, the leaf extract at 250 μg/ml and the stem extract at 100 μg/mL upregulated the expression of IL-10 and H1F1.
These findings indicate that Trichilia silvatica extracts exhibit notable antioxidant activity, as evidenced by greater than 75 % inhibition of DPPH radicals and elevated FRAP values. Additionally, the extracts demonstrated anti-inflammatory properties by downregulating key pro-inflammatory mediators, including TNF-α, NF-κB, and COX-2, while upregulating the anti-inflammatory cytoline IL-10 and enhancing enhancing tissue oxygenation and nutrient supply through increased expression of HIF-1. These effects highlight the potential of T. silvatica extracts as therapeutic agents for managing inflammatory diseases associated with oxidative stress, thereby supporting their traditional medicinal use.
楝科植物，如被称为“catiguá-branco”的Trichilia silvatica C. DC.，由于其多样且重要的次生代谢产物，在植物化学研究中引起了相当大的关注。Trichilia silvatica传统上被用于巴西医学治疗炎症性疾病。此外，研究报告了其抗氧化和抗菌特性，突出了其潜在的治疗应用。
本研究旨在评估Trichilia silvatica叶和茎提取物在RAW264.7巨噬细胞暴露于脂多糖（LPS）或过氧化氢（HO）后调节氧化炎症的潜力，并阐明其潜在的作用机制。
使用薄层色谱（TLC）、配备反相Hypersil C-18柱的高效液相色谱（HPLC）和分光光度法对提取物的植物化学成分进行表征。使用2,2-二苯基-1-苦基肼（DPPH）和铁还原抗氧化能力（FRAP）测定法评估其抗氧化活性。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法评估细胞活力，同时测定提取物处理并随后用HO刺激的细胞中的过氧化氢酶（CAT）和超氧化物歧化酶（SOD）活性以及一氧化氮（NO）生成。使用逆转录定量聚合酶链反应（RT-qPCR）定量核因子κB（NF-κB）、环氧化酶2（COX-2）、肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）和缺氧诱导因子-1（HIF-1）的基因表达水平。
Trichilia silvatica提取物显示含有萜类/类固醇、香豆素、缩合单宁和酚酸，包括绿原酸和咖啡酸。结果表明，100μg/ml的叶提取物以及100μg/ml和250μg/ml的茎提取物可保持或提高细胞活力，赋予对HO诱导的氧化应激的保护作用。这些浓度显著增加了CAT活性，而SOD活性未受影响。当用100μg/ml和250μg/ml的叶和茎提取物处理细胞时，一氧化氮生成显著减少。此外，FRAP值显示在250μg/mL时抗氧化能力增加。100μg/mL和250μg/mL的叶和茎提取物均表现出超过75%的DPPH自由基清除能力，并下调了包括NF-κB、TNF-α和COX-2在内的促炎细胞因子的表达。值得注意的是，250μg/ml的叶提取物和100μg/mL的茎提取物上调了IL-10和H1F1的表达。
这些发现表明，Trichilia silvatica提取物表现出显著的抗氧化活性，DPPH自由基抑制率大于75%和FRAP值升高证明了这一点。此外，提取物通过下调关键促炎介质，包括TNF-α、NF-κB和COX-2，表现出抗炎特性，同时上调抗炎细胞因子IL-10，并通过增加HIF-1的表达增强组织氧合和营养供应。这些作用突出了Trichilia silvatica提取物作为治疗与氧化应激相关的炎症性疾病的治疗剂的潜力，从而支持了它们的传统药用用途。
