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κ-卡拉胶寡糖及其硫酸化衍生物对脂多糖激活的小胶质细胞的免疫调节作用。

The immune regulation of κ-carrageenan oligosaccharide and its desulfated derivatives on LPS-activated microglial cells.

机构信息

Bioengineering College of Dalian University, Dalian 116622, China.

出版信息

Neurochem Int. 2012 Oct;61(5):689-96. doi: 10.1016/j.neuint.2012.06.019. Epub 2012 Jul 3.

Abstract

Neurodegenerative disease involves an inflammatory response in the central nervous system characterized by an increase in inflammatory cytokines and activation of microglial cell. To reveal the immune regulation activity of κ-carrageenan oligosaccharides (KOS) on microglia cell activated by LPS and the relationship between the sulfate group content of KOS and its immune regulation activity, KOS was prepared by enzymatic hydrolysis. The degradation products of κ-carrageenan were analyzed by high performance liquid chromatography (HPLC), ESI-TOF-MS and (13)C NMR spectroscopy, and the results indicated that the hydrolyzed products of the κ-carrageenase were κ-neocarrabiose-sulfate, κ-neocarrahexaose-sulfate and κ-neocarraoctaose-sulfate, respectively. Then desulfated derivatives of KOS (DSK) were obtained with DMSO-methanol-pyridine method. The effect of KOS and DSK on the viability of microglia cell activated by LPS was determined with MTT method. Griess assay and ELISA method were used to determine the contents of NO/NO(2-), TNF-α and IL-10 released by activated microglia cell, respectively. The results showed that KOS could inhibit the viability and content of NO, TNF-α and IL-10 released by LPS-activated microglia cell dose dependently. Compared with that of KOS, the inhibiting activity of DSK is weaker. So it could be concluded that KOS could protect microglial cell from being activated by LPS, and there was a positive relationship between the sulfate group content of KOS and its protection function.

摘要

神经退行性疾病涉及中枢神经系统中的炎症反应,其特征在于炎症细胞因子的增加和小胶质细胞的激活。为了揭示κ-卡拉胶低聚糖(KOS)对 LPS 激活的小胶质细胞的免疫调节活性及其硫酸基含量与免疫调节活性之间的关系,通过酶解制备 KOS。通过高效液相色谱(HPLC)、ESI-TOF-MS 和(13)C NMR 光谱分析 κ-卡拉胶的降解产物,结果表明 κ-卡拉胶酶的水解产物分别为κ-新卡拉双糖硫酸盐、κ-新卡拉六糖硫酸盐和 κ-新卡拉八糖硫酸盐。然后用 DMSO-甲醇-吡啶法得到 KOS 的脱硫酸衍生物(DSK)。用 MTT 法测定 KOS 和 DSK 对 LPS 激活的小胶质细胞活力的影响。用格里斯测定法和 ELISA 法分别测定激活的小胶质细胞释放的 NO/NO(2-)、TNF-α和 IL-10 的含量。结果表明,KOS 可剂量依赖性地抑制 LPS 激活的小胶质细胞的活力和 NO、TNF-α和 IL-10 的含量。与 KOS 相比,DSK 的抑制活性较弱。因此,可以得出结论,KOS 可以保护小胶质细胞免受 LPS 的激活,并且 KOS 的硫酸基含量与其保护功能之间存在正相关关系。

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