Department of Biochemistry, Faculty of Science, University of Dschang, P. O. Box 67, Dschang, Cameroon; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Rue Michel-Servet 1, 1211, Geneva, Switzerland; School of Pharmaceutical Sciences, University of Geneva, Rue Michel-Servet 1, 1211, Geneva, Switzerland.
Department of Organic Chemistry, Faculty of Sciences, University of Yaoundé I., P.O. Box 812, Yaoundé, Cameroon.
J Ethnopharmacol. 2021 Mar 25;268:113637. doi: 10.1016/j.jep.2020.113637. Epub 2020 Nov 29.
Ocimum gratissimum is a plant spice widely used in African traditional medicine to treat pain-related conditions. However, the anti-inflammatory mechanisms underlying this activity and the main active ingredients in O. gratissimum have not yet been fully characterized.
To isolate and identify the main anti-inflammatory active constituents of Ocimum gratissimum extract and their underlying mechanisms in murine macrophages.
Chromatographic techniques and spectroscopic data were used for compounds isolation and identification. Inflammatory conditions were produced in cultured RAW 264.7 macrophage cells by the application of lipopolysaccharide (LPS). The WST-1 assay was used to evaluate the cell viability, and the nitric oxide production was quantified by the Griess reagent method. The fluorometric cyclooxygenase (COX) activity assay kit was used to assess the activity of COX-1 and COX-2 enzymes. The levels of IFN-γ, TNF-α, IL-2, IL-4, IL-6, and IL-10 cytokines and the apoptosis-inducing effect were measured by flow cytometer using the cytometric Bead Array (CBA) Human Th1/Th2 Cytokine Kit II and FITC Annexin V Apoptosis Detection kit, respectively.
The results showed that the extract and fractions of Ocimum gratissimum inhibit nitric oxide production and the proliferation of Raw 264.7 macrophage cells. The bioguided fractionation led to the identification of pentacyclic triterpenes as anti-inflammatory bioactive compounds. Pomolic and tormentic acids being the most active, inhibiting the secretion of IFN-γ cytokine, COX enzyme, and inducing apoptosis in activated Raw 264.7 macrophage cells.
This study revealed that pomolic and tormentic acids are the main active principles responsible at least in part for the anti-inflammatory effect of the extract of Ocimum gratissimum. Besides of providing more evidence for the traditional use of Ocimum gratissimum against inflammatory disorders, this study reveals the multitarget potential of pomolic and tormentic acids as promising future drugs against inflammatory diseases.
奥图姆 gratissimum 是一种植物香料，广泛用于非洲传统医学治疗与疼痛相关的疾病。然而，这种活性的抗炎机制和奥图姆 gratissimum 的主要活性成分尚未得到充分描述。
分离和鉴定奥图姆 gratissimum 提取物的主要抗炎活性成分及其在鼠巨噬细胞中的作用机制。
采用色谱技术和光谱数据对化合物进行分离和鉴定。在培养的 RAW 264.7 巨噬细胞中，通过应用脂多糖（LPS）产生炎症条件。使用 WST-1 测定法评估细胞活力，并用 Griess 试剂法定量测定一氧化氮的产生。使用荧光环氧化酶（COX）活性测定试剂盒评估 COX-1 和 COX-2 酶的活性。通过流式细胞术使用细胞因子分析试剂盒 II 和 FITC 膜联蛋白 V 凋亡检测试剂盒分别测量 IFN-γ、TNF-α、IL-2、IL-4、IL-6 和 IL-10 细胞因子的水平和诱导凋亡的作用。
结果表明，奥图姆 gratissimum 的提取物和馏分抑制一氧化氮的产生和 Raw 264.7 巨噬细胞的增殖。生物导向的分离导致鉴定出五环三萜类化合物为抗炎生物活性化合物。pomolic 和 tormentic 酸是最活跃的，抑制 IFN-γ细胞因子、COX 酶的分泌，并诱导活化的 Raw 264.7 巨噬细胞凋亡。
本研究表明，pomolic 和 tormentic 酸是负责奥图姆 gratissimum 提取物抗炎作用的主要活性成分。除了为奥图姆 gratissimum 传统用于治疗炎症性疾病提供更多证据外，本研究还揭示了 pomolic 和 tormentic 酸作为治疗炎症性疾病的潜在多靶药物的潜力。
