Key Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan 750021, China.
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Molecules. 2020 Mar 16;25(6):1351. doi: 10.3390/molecules25061351.
L. is a widely used functional food and medicinal herb in Asian countries. polysaccharides (LBP) are considered as one of the major medicinal components of fruit and exhibits a wide range of biological activities. Here, we investigated the immunomodulatory effects of LBP and its uptake behaviors at the cellular level. LBP was prepared by water extraction and ethanol precipitation, and divided into two fractions based on the molecular weight distribution by ultrafiltration (LBP > 10 kDa and LBP < 10 kDa). The physicochemical properties of LBP and LBP fractions were well characterized. The LBP > 10 kDa fraction greatly enhanced the viability of macrophages RAW264.7 cells and induced cell polarization, but had weak effects to other tested tumor cell lines and normal cell line. This fraction could regulate the production of NO, TNF-α, IL-6 and ROS in RAW264.7 cells, suggesting both pro-inflammatory and anti-inflammatory effects. The dye-labeled LBP could be internalized into all tested cell lines and accumulated in lysosomes. The internalization of LBP in RAW264.7 cells is mainly through the clathrin-mediated endocytosis pathway. The Caco-2 intestinal transport experiment demonstrated that the dye labeled LBP could be transported through the Caco-2 cell monolayer (mimic intestinal epithelium) through clathrin-mediated endocytosis. These results demonstrate the immunomodulatory effects of LBP and its effective uptake by macrophages and intestine.
灵芝是亚洲国家广泛使用的功能性食品和药用草本植物。灵芝多糖（LBP）被认为是灵芝的主要药用成分之一，具有广泛的生物活性。在这里，我们研究了 LBP 的免疫调节作用及其在细胞水平上的摄取行为。LBP 通过水提和乙醇沉淀制备，并通过超滤（LBP>10 kDa 和 LBP<10 kDa）根据分子量分布分为两部分。LBP 和 LBP 级分的理化性质得到了很好的表征。LBP>10 kDa 级分大大提高了巨噬细胞 RAW264.7 细胞的活力并诱导细胞极化，但对其他测试的肿瘤细胞系和正常细胞系的影响较弱。该级分可调节 RAW264.7 细胞中 NO、TNF-α、IL-6 和 ROS 的产生，提示具有促炎和抗炎作用。标记染料的 LBP 可被内化到所有测试的细胞系中，并在溶酶体中积累。LBP 在 RAW264.7 细胞中的内化主要通过网格蛋白介导的内吞作用途径。Caco-2 肠转运实验表明，标记染料的 LBP 可以通过 Caco-2 细胞单层（模拟肠上皮）通过网格蛋白介导的内吞作用进行转运。这些结果表明 LBP 具有免疫调节作用，并且可以被巨噬细胞和肠道有效摄取。
