Department of Food Science and Biotechnology, National Chung Hsing University, Taichung 40227, Taiwan, Republic of China (ROC).
J Ethnopharmacol. 2013 May 2;147(1):164-73. doi: 10.1016/j.jep.2013.02.028. Epub 2013 Feb 28.
Lotus plumule is widely used as traditional Chinese medicine. Among the active components in lotus plumule, polysaccharides exhibit promising potential for its potent anti-inflammatory effects. However, the anti-inflammatory mechanism of purified polysaccharides from lotus plumule remains unknown. To evaluate their anti-inflammatory potential and possible mechanisms of purified polysaccharides in lotus plumule, two active lotus plumule polysaccharides, fractions F1 and F2, were subjected to assay their anti-inflammatory potential and possible mechanisms using murine primary splenocytes in the absence or presence of lipopolysaccharide (LPS).
Two purified active lotus plumule polysaccharides, F1 and F2, were cultured independently with murine primary splenocytes in the absence or presence of LPS under four different experiment models in vitro. Changes in pro-inflammatory IL-1β, IL-6 and TNF-α, as well as anti-inflammatory IL-10 cytokines secreted by the treated splenocytes were determined using an enzyme-linked immunosorbent assay (ELISA). The amount of toll-like receptor (TLR)-2 and TLR-4 mRNA expression levels in the cells were quantitated using a two-step real-time polymerase chain reaction (PCR) assay.
The results showed that F1 and F2 treatments alone, particularly F2, significantly (P<0.05) decreased pro-/anti-inflammatory (IL-1β/IL-10 and TNF-α/IL-10) cytokine secretion ratios dose-dependently. F1 and F2 treatments in the presence of LPS significantly decreased TLR-2 and/or TLR-4 mRNA expression levels in the splenocytes under inflammatory and repair experiment models.
The present study proved that F1 and F2 had strong anti-inflammatory effects through inhibiting TLR-2 and/or TLR-4 expressions in the splenocytes in normal, inflammatory and repair situations. Our results further suggest that F2, which is a glycoprotein with low molecular weight of 25.7 kDa, may serve as a promising lead for the development of selective TLR antagonistic agents for inflammatory diseases.
莲心是一种广泛使用的传统中药。在莲心的活性成分中，多糖具有很强的抗炎作用，具有很大的应用潜力。然而，莲心纯化多糖的抗炎机制尚不清楚。为了评估其抗炎潜力及其在无脂多糖（LPS）或存在 LPS 时莲心纯化多糖的可能机制，用小鼠原代脾细胞在体外四种不同的实验模型中分别培养两种活性莲心多糖 F1 和 F2，以检测其抗炎潜力及其可能的机制。
两种纯化的活性莲心多糖 F1 和 F2 在无 LPS 或存在 LPS 下分别与小鼠原代脾细胞在体外四种不同的实验模型中进行培养。用酶联免疫吸附试验（ELISA）测定经处理的脾细胞分泌的促炎细胞因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）以及抗炎细胞因子白细胞介素-10（IL-10）的变化。用两步实时聚合酶链反应（PCR）法定量细胞中 toll 样受体（TLR）-2 和 TLR-4 mRNA 表达水平。
结果表明，F1 和 F2 单独处理，特别是 F2，显著（P<0.05）降低了促炎/抗炎（IL-1β/IL-10 和 TNF-α/IL-10）细胞因子分泌比，呈剂量依赖性。在炎症和修复实验模型中，F1 和 F2 在 LPS 存在下处理，显著降低了脾细胞中 TLR-2 和/或 TLR-4 mRNA 表达水平。
本研究证明，F1 和 F2 通过抑制正常、炎症和修复情况下脾细胞中的 TLR-2 和/或 TLR-4 表达，具有很强的抗炎作用。我们的结果进一步表明，F2 是一种具有 25.7 kDa 低分子量的糖蛋白，可能成为开发用于炎症性疾病的选择性 TLR 拮抗剂的有前途的先导化合物。
