Heteroglycan of Annona muricata leaves: Extraction, characterization, modulation of the inflammatory response in raw 264.7 macrophages and peripheral Blood mononuclear cells

Judith Emery Ngomoyogoli Kanemoto , Fils Armand Ella , Alice Louise Woguia , Stanley Olivier Kanemoto , Thaddée Boudjeko , Judith Laure Ngondi , Nicolas Yanou Njintang
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Abstract

The natural products contained in Annona muricata leaves are characterized by their ability to effectively enhance the immune response without exhibiting toxicity. The immunomodulatory effect of natural polysaccharides is considered one of the most important bioactive functions. In this study, we investigated the antioxidant activities and immunological properties of heteroglycans extracted from A. muricata leaves using hot water followed by ethanol precipitation. The polysaccharide was characterized for its physicochemical properties (UV spectroscopy, FTIR, gas chromatography), antioxidant potential (scavenging properties, chelating ability, reducing capability, and protection against DNA damage), immunomodulatory activities, and production of pro-inflammatory cytokines using peripheral blood mononuclear cells (PBMCs) and RAW 264.7 cells. The results indicated that the heteroglycans consisted of Glu-Man-GalA-GluA-Xyl and GluA-Man-Glu, respectively, for low and high molecular weight water-soluble polysaccharides coded as LWPC and HWPC. They exhibited strong metal chelating and scavenging activities. Additionally, each sample provided protection against DNA damage induction, with LWPC exhibiting up to 73.1 % protection. LWPC and HWPC significantly down-regulated the expression of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and COX-2, as well as NO production. At 200 µg/mL, only LWPC increased the production of IL-1β and IL-6 in PBMCs. Therefore, LWPC and HWPC hold promise for inhibiting radical-induced and inflammatory disorders.

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鼠尾草叶杂聚糖:提取、表征、调节生 264.7 巨噬细胞和外周血单核细胞的炎症反应
鼠尾草叶所含天然产品的特点是能够有效增强免疫反应,且无毒性。天然多糖的免疫调节作用被认为是最重要的生物活性功能之一。在这项研究中,我们用热水和乙醇沉淀法研究了从鼠尾草叶中提取的杂多糖的抗氧化活性和免疫学特性。研究人员利用外周血单核细胞(PBMCs)和 RAW 264.7 细胞对多糖的理化性质(紫外光谱、傅立叶变换红外光谱、气相色谱法)、抗氧化潜力(清除特性、螯合能力、还原能力和 DNA 损伤保护)、免疫调节活性和促炎细胞因子的产生进行了表征。结果表明,杂聚多糖由 Glu-Man-GalA-GluA-Xyl 和 GluA-Man-Glu 组成,分别为低分子量和高分子量水溶性多糖,代号分别为 LWPC 和 HWPC。它们具有很强的金属螯合和清除活性。此外,每种样品都能保护 DNA 免受损伤,其中 LWPC 的保护率高达 73.1%。LWPC 和 HWPC 能显著降低促炎细胞因子 TNF-α、IL-1β、IL-6 和 COX-2 的表达以及 NO 的产生。在 200 µg/mL 的浓度下,只有 LWPC 能增加 PBMC 中 IL-1β 和 IL-6 的产生。因此,LWPC和HWPC有望抑制自由基引起的炎症性疾病。
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