Physicochemical characterization of polysaccharide from the leaf of Dendrobium officinale and effect on LPS induced damage in GES-1 cell
Physicochemical characterization of polysaccharide from the leaf of Dendrobium officinale and effect on LPS induced damage in GES-1 cell
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铁皮石斛叶多糖的理化特性及其对LPS诱导的GES-1细胞损伤的影响
DOI：
10.1016/j.ijbiomac.2020.01.026
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发表时间：
2020
影响因子：
8.2
通讯作者：
Suhong Chen
中科院分区：
文献类型：
--
作者：
Ke Yang;Tingting Lu;Lianghui Zhan;Cong Zhou;Ningyu Zhang;Shanshan Lei;Yuzhi Wang;Junsheng Yang;Meiqiu Yan;Guiyuan Lv;Suhong Chen
首先从铁皮石斛叶片中通过热水提取和醇沉分离得到该多糖，并用DEAE-52和Sephadex G-100层析对其进行纯化。采用HPLC、GPC、FT-IR和NMR等方法对LDOP-1的结构进行表征，并分析其对lps诱导的GES-1细胞损伤的保护作用。结果表明，LDOP为均匀多糖，平均分子量为91.8 kDa，由Man、Gla、Glc、Glc酸和Ara组成，摩尔比为2.0:1.3:1.6:1.7:0.7。LDOP具有1,6-linked α-d-Glup和1,4-linked α-d-Manp两种残基类型。活性研究表明，ldp -1能显著抑制lps诱导的gus -1细胞损伤中肿瘤坏死因子-α (TNF-α)、白细胞介素-1β (IL-1β)和IL-6的释放，降低TLR4、磷酸化-NF-κB、ASC、NLRP3、裂解-IL-1β、IL-6和Bax的蛋白表达，提高Bcl-2蛋白表达，下调裂解caspase-1 / pro-caspase-1、磷酸化-i -κB α / i -κ α、磷酸化-NF-кB / NF-κB的比值。这些结果强烈提示，LDOP可以通过抑制TLR4/NF-κB信号通路调节的炎症细胞因子的释放来预防lps诱导的GES-1细胞损伤。
The polysaccharide was first successfully isolated from the leaf ofDendrobium officinaleby hot water extraction and alcohol precipitation and further purified using DEAE-52 and Sephadex G-100 chromatography. The structure of LDOP-1 was characterized by HPLC, GPC, and FT-IR and NMR spectroscopy, and its protective effect on LPS-induced GES-1 cell injury was analyzed. Results showed that LDOP was a homogeneous polysaccharide with average molecular weight of 91.8 kDa and consisted of Man, Gla, Glc, Glc acid, and Ara at a molar ratio of 2.0:1.3:1.6:1.7:0.7. LDOP had two types of residues, including 1,6-linked α-d-Glup and 1,4-linked α-d-Manp. Activity studies indicated that LDOP-1 can significantly suppress the release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 from LPS-induced GES-1 cell injury, decreased the protein expressions of TLR4, phospho-NF-κB, ASC, NLRP3, cleaved-IL-1β, IL-6, and Bax, increased the protein expression of Bcl-2, and downregulated the ratios of cleaved caspase-1 to pro-caspase-1, phospho-IκBα to IκBα, and phospho-NF-кB to NF-κB. These findings strongly suggested that LDOP can prevent LPS-induced GES-1 cell injury by inhibiting the release of inflammatory cytokines regulated via the TLR4/NF-κB signal pathways.