Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, School of Food Science and Pharmaceutics, Zhejiang Ocean University, Zhoushan, Zhejiang 316022, China.
Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, School of Food Science and Pharmaceutics, Zhejiang Ocean University, Zhoushan, Zhejiang 316022, China.
Int J Biol Macromol. 2019 Aug 15;135:203-211. doi: 10.1016/j.ijbiomac.2019.05.082. Epub 2019 May 17.
The purpose of this study was to investigate structure of Hemp seed polysaccharide (HSP) and the protective effect of HSP from HO-induced oxidative damage in IPEC-1 cells and the possible mechanism of this protection. Analysis of monosaccharide composition and structure of two fractions HSP and HSP from polysaccharide of Hemp seed (HSPc) were analyzed by high performance liquid chromatography (HPLC) and Fourier transform infrared spectroscopy (FT-IR). The results showed that both HSP and HSP contain sulfate groups, which are sulfated polysaccharides. In IPEC-1 cells model, the release of LDH and MDA was significantly decreased, and the activities of SOD, GSH-Px and CAT were significantly increased in HSP and HSP-treated group. HSP dramatically increased the gene expression of antioxidant enzymes and phase II detoxification enzymes measured by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). In addition, HSP up-regulated the expression level of intracellular transcription factor Nuclear factor erythroid-2-related factor 2 (Nrf2) and inhibited the level of Kelch-like ECH-associated protein 1 (Keap1) with Western blot analysis. Collectively, the present study suggested that HSP has the protective effect of IPEC-1 cells against HO-induecd oxidative stress. This protection mechanism may be related to activation of the Keap1/Nrf2 signaling pathway.
本研究旨在探讨麻籽多糖(HSP)的结构及其对 HO 诱导的 IPEC-1 细胞氧化损伤的保护作用及可能的保护机制。采用高效液相色谱(HPLC)和傅里叶变换红外光谱(FT-IR)分析两种麻籽多糖 HSP 和 HSPc 的单糖组成和结构。结果表明,HSP 和 HSP 均含有硫酸基团,属于硫酸化多糖。在 IPEC-1 细胞模型中,HSP 和 HSP 处理组可显著降低 LDH 和 MDA 的释放,显著提高 SOD、GSH-Px 和 CAT 的活性。实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测结果表明,HSP 可显著上调抗氧化酶和Ⅱ相解毒酶相关基因的表达。此外,Western blot 分析结果表明,HSP 可上调细胞内转录因子核红细胞 2 相关因子 2(Nrf2)的表达水平,抑制 Kelch 样 ECH 相关蛋白 1(Keap1)的水平。综上所述,本研究表明 HSP 对 HO 诱导的 IPEC-1 细胞氧化应激具有保护作用。这种保护机制可能与 Keap1/Nrf2 信号通路的激活有关。