School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Zhejiang Shouxiangu Institute of Rare Medicine Plant, Wuyi, 321200, China.
J Immunol Res. 2021 Mar 5;2021:6696606. doi: 10.1155/2021/6696606. eCollection 2021.
has certain components with known pharmacological effects, including strengthening immunity and anti-inflammatory activity. seeds inherit all its biological characteristics. spore polysaccharide (GLSP) is the main active ingredient to enhance these effects. However, its specific biological mechanisms are not exact. Our research is aimed at revealing the specific biological mechanism of GLSP to enhance immunity and inhibit the growth of H22 hepatocellular carcinoma cells.
We extracted primary macrophages (M) from BALB/c mice and treated them with GLSP (800 g/mL, 400 g/mL, and 200 g/mL) to observe its effects on macrophage polarization and cytokine secretion. We used GLSP and GLSP-intervened macrophage supernatant to treat H22 tumor cells and observed their effects using MTT and flow cytometry. Moreover, real-time fluorescent quantitative PCR and western blotting were used to observe the effect of GLSP-intervened macrophage supernatant on the PI3K/AKT and mitochondrial apoptosis pathways.
In this study, GLSP promoted the polarization of primary macrophages to M1 type and the upregulation of some cytokines such as TNF-, IL-1, IL-6, and TGF-1. The MTT assay revealed that GLSP+M at 400 g/mL and 800 g/mL significantly inhibited H22 cell proliferation in a dose-dependent manner. Flow cytometry analysis revealed that GLSP+Mø induced apoptosis and cell cycle arrest at the G2/M phase, associated with the expression of critical genes and proteins (PI3K, p-AKT, BCL-2, BAX, and caspase-9) that regulate the PI3K/AKT pathway and apoptosis. GLSP reshapes the tumor microenvironment by activating macrophages, promotes the polarization of primary macrophages to M1 type, and promotes the secretion of various inflammatory factors and cytokines.
Therefore, as a natural nutrient, GLSP is a potential agent in hepatocellular carcinoma cell treatment and induction of apoptosis.
具有某些具有已知药理作用的成分，包括增强免疫力和抗炎活性。 种子继承了其所有的生物特征。 孢子多糖（GLSP）是增强这些作用的主要活性成分。然而，其具体的生物学机制尚不清楚。我们的研究旨在揭示 GLSP 增强免疫力和抑制 H22 肝癌细胞生长的具体生物学机制。
我们从 BALB/c 小鼠中提取原代巨噬细胞（M），并用 GLSP（800μg/mL、400μg/mL 和 200μg/mL）处理，观察其对巨噬细胞极化和细胞因子分泌的影响。我们用 GLSP 和 GLSP 干预的巨噬细胞上清液处理 H22 肿瘤细胞，并用 MTT 和流式细胞术观察其作用。此外，实时荧光定量 PCR 和 Western blot 用于观察 GLSP 干预的巨噬细胞上清液对 PI3K/AKT 和线粒体凋亡途径的影响。
在这项研究中，GLSP 促进了原代巨噬细胞向 M1 型极化，并上调了 TNF-α、IL-1、IL-6 和 TGF-β1 等一些细胞因子。MTT 测定显示，GLSP+M 在 400μg/mL 和 800μg/mL 时以剂量依赖的方式显著抑制 H22 细胞增殖。流式细胞术分析显示，GLSP+Mø 诱导凋亡和细胞周期停滞在 G2/M 期，与调节 PI3K/AKT 途径和凋亡的关键基因和蛋白（PI3K、p-AKT、BCL-2、BAX 和 caspase-9）的表达有关。GLSP 通过激活巨噬细胞重塑肿瘤微环境，促进原代巨噬细胞向 M1 型极化，并促进各种炎症因子和细胞因子的分泌。
因此，作为一种天然营养素，GLSP 是肝癌细胞治疗和诱导细胞凋亡的潜在药物。
