Experiential Master of Science in Biotechnology, College of Science, Northeastern University, Boston, MA, USA.
LAQUA (Laboratório de Química da Unesp Assis), University of São Paulo State (UNESP), Assis, SP, Brazil.
BMC Complement Med Ther. 2023 Oct 7;23(1):356. doi: 10.1186/s12906-023-04190-7.
Carboxymethylated Lasiodiplodan (LaEPS-C), Lasiodiplodia theobromae β-glucan exopolysaccharide derivative, has a well-known range of biological activities. Compared to LaEPS-C, its fractions, Linear (LLaEPS-C) and Branched (BLaEPS-C), have biological potentialities scarcely described in the literature. So, in this study, we investigate the immunomodulatory, antiviral, antiproliferative, and anticoagulant activities of LLaEPS-C and BLaEPS-C and compare them to the LaEPS-C.
LaEPS was obtained from L. theobromae MMBJ. After carboxymethylation, LaEPS-C structural characteristics were confirmed by Elementary Composition Analysis by Energy Dispersive X-Ray Detector (EDS), Fourier Transform Infrared (FTIR), and Nuclear Magnetic Resonance (NMR). The immunomodulatory activity on cytokine secretion was evaluated in human monocyte-derived macrophage cultures. The antiviral activity was evaluated by Hep-2 cell viability in the presence or absence of hRSV (human respiratory syncytial virus). In vitro antiproliferative activity was tested by sulforhodamine B assay. The anticoagulant activity was determined by APTT (Activated Partial Thromboplastin Time) and PT (Prothrombin Time).
LaEPS-C showed low macrophage cell viability only at 100 µg/mL (52.84 ± 24.06, 48 h), and LLaEPS-C presented no effect. Conversely, BLaEPS-C showed cytotoxicity from 25 to 100 µg/mL (44.36 ± 20.16, 40.64 ± 25.55, 33.87 ± 25.16; 48 h). LaEPS-C and LLaEPS-C showed anti-inflammatory activity. LaEPS-C presented this at 100 µg/mL (36.75 ± 5.53, 48 h) for IL-10, and LLaEPS-C reduces TNF-α cytokine productions at 100 µg/mL (18.27 ± 5.80, 48 h). LLaEPS-C showed an anti-hRSV activity (0.7 µg/ml) plus a low cytotoxic activity for Hep-2 cells (1.4 µg/ml). LaEPS-C presented an antiproliferative activity for NCI-ADR/RES (GI 65.3 µg/mL). A better PT was achieved for LLaEPS-C at 5.0 µg/mL (11.85 ± 0.87s).
These findings demonstrated that carboxymethylation effectively improves the biological potential of the LaEPS-C and their fractions. From those polysaccharides tested, LLaEPS provided the best results with low toxicity for anti-inflammatory, antiviral, and anticoagulant activities.
羧甲基化 Lasiodiplodan（LaEPS-C）是胶孢炭疽菌β-葡聚糖外多糖衍生物，具有众所周知的一系列生物活性。与 LaEPS-C 相比，其馏分线性（LLaEPS-C）和支链（BLaEPS-C）具有文献中鲜有描述的生物潜力。因此，在这项研究中，我们研究了 LLaEPS-C 和 BLaEPS-C 的免疫调节、抗病毒、抗增殖和抗凝活性，并将其与 LaEPS-C 进行了比较。
从胶孢炭疽菌 MMBJ 中获得 LaEPS。羧甲基化后，通过能量色散 X 射线探测器（EDS）、傅里叶变换红外（FTIR）和核磁共振（NMR）对 LaEPS-C 的结构特征进行了确认。在人单核细胞衍生的巨噬细胞培养物中评价细胞因子分泌的免疫调节活性。通过 Hep-2 细胞活力评估抗病毒活性，存在或不存在 hRSV（人呼吸道合胞病毒）。通过磺酰罗丹明 B 测定法测试体外抗增殖活性。通过 APTT（活化部分凝血活酶时间）和 PT（凝血酶原时间）测定法测定抗凝活性。
LaEPS-C 仅在 100μg/mL（48 小时时 52.84±24.06）时对巨噬细胞存活率表现出低毒性，而 LLaEPS-C 则没有影响。相反，BLaEPS-C 从 25 到 100μg/mL（48 小时时 44.36±20.16、40.64±25.55、33.87±25.16）时表现出细胞毒性。LaEPS-C 和 LLaEPS-C 表现出抗炎活性。LaEPS-C 在 100μg/mL（48 小时时 36.75±5.53）时表现出 IL-10 活性，而 LLaEPS-C 则在 100μg/mL（48 小时时 18.27±5.80）时降低 TNF-α细胞因子的产生。LLaEPS-C 对 hRSV 表现出抗病毒活性（0.7μg/ml），对 Hep-2 细胞的细胞毒性较低（1.4μg/ml）。LaEPS-C 对 NCI-ADR/RES（GI 65.3μg/ml）表现出抗增殖活性。LLaEPS-C 在 5.0μg/mL（11.85±0.87s）时可获得更好的 PT。
这些发现表明羧甲基化可有效提高 LaEPS-C 及其馏分的生物学潜力。在所测试的多糖中，LLaEPS 具有最佳的抗炎、抗病毒和抗凝活性，且毒性较低。
