Department of Biology & Guangdong Provincial Key Laboratory of Marine Biotechnology, Institute of Marine Sciences, College of Science, Shantou University, Shantou 515063, China.
Mar Drugs. 2020 Oct 28;18(11):538. doi: 10.3390/md18110538.
In this study, (S.p.) polysaccharide (PSP) was obtained by ultrasonic-microwave-assisted extraction (UMAE) and purified by an aqueous two-phase system (ATPS). Two different methods were applied to purified (S.p.) polysaccharide (PSP), respectively, due to PSP as a complex multi-component system. Three polysaccharide fractions (PSP-1, PSP-2, and PSP-3) with different acidic groups were obtained after PSP was fractionated by the diethyl aminoethyl (DEAE)-52 cellulose chromatography, and two polysaccharide fractions (PSP-L and PSP-H) with different molecular weight were obtained by ultrafiltration centrifugation. The chemoprotective effects of PSP in cyclophosphamide (Cy) treated mice were investigated. The results showed that PSP could significantly increase spleen and thymus index, peripheral white blood cells (PWBC), and peripheral blood lymphocytes (PBL). The in vivo immunostimulatory assays demonstrated that PSP could in dose-dependent increase of TNF-α, IL-10, and IFN-γ production in sera. The in vitro immunostimulatory assays showed that PSP and its fractions (PSPs) could evidently enhance the proliferation of splenocytes and RAW 264.7 cells and increase the productions of nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6). PSPs could also enhance phagocytic activity of RAW 264.7 cells. The acidic polysaccharide fractions of PSP-2, PSP-3, and PSP-L with small molecular weight had the higher immunostimulatory activity. Signaling pathway research results indicated that PSP-L activated RAW264.7 cells through MAPKs, NF-κB signaling pathways via TLR4 receptor.
在这项研究中,(S.p.)多糖(PSP)通过超声微波辅助提取(UMAE)获得,并通过双水相体系(ATPS)纯化。由于 PSP 是一种复杂的多组分体系,因此分别应用了两种不同的方法来纯化(S.p.)多糖(PSP)。PSP 通过 DEAE-52 纤维素色谱分离后,得到三个具有不同酸性基团的多糖级分(PSP-1、PSP-2 和 PSP-3),超滤离心得到两个具有不同分子量的多糖级分(PSP-L 和 PSP-H)。研究了 PSP 在环磷酰胺(Cy)处理的小鼠中的化学保护作用。结果表明,PSP 可显著增加脾和胸腺指数、外周白细胞(PWBC)和外周血淋巴细胞(PBL)。体内免疫刺激试验表明,PSP 可剂量依赖性增加血清中 TNF-α、IL-10 和 IFN-γ的产生。体外免疫刺激试验表明,PSP 及其级分(PSPs)可明显增强脾细胞和 RAW 264.7 细胞的增殖,并增加一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)和白细胞介素 6(IL-6)的产生。PSPs 还可以增强 RAW 264.7 细胞的吞噬活性。具有小分子量的酸性多糖级分 PSP-2、PSP-3 和 PSP-L 具有更高的免疫刺激活性。信号通路研究结果表明,PSP-L 通过 TLR4 受体激活 RAW264.7 细胞通过 MAPKs、NF-κB 信号通路。
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