| 林丽远,唐燕飞,杨胜涛,等.等鞭金藻多肽IEC对脂多糖诱导血管内皮炎症的保护作用研究[J].中国海洋药物,2022,41(3):50-56. |
| 等鞭金藻多肽IEC对脂多糖诱导血管内皮炎症的保护作用研究 |
| The Protective Effect of Polypeptide IEC from Isochrysis zhangjiang on LPS induced vascular endonal inflammation |
| 投稿时间：2021-06-27 修订日期：2021-10-05 |
| DOI： |
| 中文关键词: 等鞭金藻 多肽 HUVEC 抗炎 脂多糖 |
| English Keywords:Isochrysis polypeptide HUVECs anti-inflammatory LPS |
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| 中文摘要: |
| 目的 探究氨基酸序列为Ile-Ile-Ala-Val-Glu-Ala-Gly-Cys的等鞭金藻多肽IEC对LPS诱导人脐静脉内皮细胞（HUVEC）炎症反应的保护作用。方法 用噻唑蓝（MTT）法确定细胞活力，DCFH-DA探针法和DAF-FM DA探针法分别测定活性氧（ROS）和NO的表达情况，蛋白免疫印迹法检测细胞间Toll样受体（TLR4）、一氧化氮合酶（NOS-2）和环氧合酶（COX-2）蛋白的表达水平，酶联免疫吸附法测定白细胞介素-6 （IL-6）和肿瘤坏死因子（TNF-α）的释放量以及分子对接分析IEC与COX-2、TLR-4蛋白的相互作用。结果 MTT法证明IEC对HUVEC细胞无明显毒性作用(P>0.05)；与对照组相比，随着实验组多肽浓度增加，可以明显降低ROS和NO的释放(P<0.05)，炎症相关细胞因子TLR-4、COX-2、NOS-2和IL-6、TNF-α的表达也明显逐渐减少(P<0.000 1)；分子对接结果表明，IEC与COX-2、TLR4能形成稳定的化学键，说明存在相互作用的可能性。结论 IEC具有良好的抑制ROS和NO生成效果，并可明显抑制TLR-4/NOS-2/COX-2信号通路的激活以及抑制炎症因子的表达，是潜在的炎症抑制剂。 |
| English Summary: |
| Objective To explore the protective effect of the Isochrysis polypeptide IEC with amino acid sequence Ile-Ile-Ala-Val-Glu-Ala-Gly-Cys on LPS-induced inflammatory response in human umbilical vein endothelial cells (HUVEC). Methods MTT method was used to determine cell viability, DCFH-DA probe method and DAF-FM DA probe method to measure the expression of reactive oxygen species (ROS) and nitric oxide (NO), respectively, and Western blotting was used to detect intercellular Toll-like receptors (TLR4), nitric oxide synthase (NOS-2) and Cyclooxygenase (COX-2) protein expression level, enzyme-linked immunosorbent assay to determine the release of interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) and molecular docking analysis IEC and COX- 2. The interaction of TLR-4 protein. Results The MTT method proved that IEC had no obvious toxic effect on HUVEC cells (P>0.05); compared with the control group, as the peptide concentration of the experimental group increased, the release of ROS and NO could be significantly reduced (P<0.05), inflammation-related cytokine TLR- 4. The expressions of COX-2, NOS-2, IL-6, and TNF-α are also significantly reduced (P<0.000 1); the molecular docking results show that IEC, COX-2 and TLR4 can form stable chemical bonds, indicating that there is a mutual Possibility of function. Conclusion IEC has a good effect of inhibiting ROS and NO generation, and can significantly inhibit the activation of TLR-4/NOS-2/COX-2 signaling pathway and inhibit the expression of inflammatory factors. It is a potential inhibitor of inflammation. |
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