| 闫成秀,于广利,张烜,等.岩藻聚糖硫酸酯F2靶向SGLT2的抗糖尿病肾病活性研究[J].中国海洋药物,2021,40(6):13-20. |
| 岩藻聚糖硫酸酯F2靶向SGLT2的抗糖尿病肾病活性研究 |
| Study on SGLT2 targeted anti-diabetic nephropathy activity of fucoidan F2 |
| 投稿时间：2021-03-31 修订日期：2021-04-30 |
| DOI： |
| 中文关键词: 岩藻聚糖硫酸酯 钠离子葡萄糖共转运蛋白2 糖尿病肾病 信号通路 |
| English Keywords:fucoidan sodium-glucose cotransporter 2 diabetic nephropathy signal pathway |
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| 中文摘要: |
| 目的 旨在从细胞水平上探讨岩藻聚糖硫酸酯F2对SGLT2的抑制作用及抗糖尿病肾病(DN)活性,并阐述其抗DN机制。方法 通过Q-Sepharose Fast Flow强阴离子交换层析法对墨角藻来源岩藻聚糖硫酸酯(FvF)进行分离纯化,并采用高效液相、离子色谱等方法测定其分子量、单糖组成、硫酸根含量等基本理化性质,利用流式细胞术,通过评价样品对SGLT2稳转细胞转运葡萄糖能力的影响,来分析样品对SGLT2的抑制活性,确定活性最优化合物。通过计算机模拟技术,探讨了岩藻寡糖与SGLT2的相互作用。采用大鼠肾系膜细胞(HBZY-1细胞),利用高糖诱导,建立HBZY-1细胞增殖模型,通过MTT法评价F2的抗DN活性,并利用Western Blot方法探究最优化合物对HBZY-1细胞模型中SphK1、NF-κB、p38、ERK1/2蛋白的影响。结果 纯化后获得了5个组分,SGLT2抑制剂筛选确定F2为最优组分,其具有分子量低、硫酸根含量高且单糖组成较为单一的特点,并且岩藻糖中硫酸根含量和聚合度影响其与SGLT2的相互作用,而2-硫酸岩藻八糖与SGLT2的亲和力最强。成功建立了HBZY-1细胞增殖模型,发现F2在较低浓度下即可发挥抗肾系膜增生作用,并且其活性随着浓度增加而增强,且F2是通过抑制SphK1、NF-κB、p38蛋白的表达,抑制ERK1/2蛋白的磷酸化来发挥抗DN活性。 |
| English Summary: |
| Objective To study fucoidan F2 about its inhibitory activity against SGLT2 along with its anti-DN activity and mechanism at the cellular level. Methods Fucoidan from Fucus vesiculosus (FvF) was purified by Q-Sepharose Fast Flow strong anion exchange chromatography, and their basic physical and chemical properties including molecular weight, monosaccharide composition and sulfate content were elucidated by high performance liquid chromatography and ion chromatography. In order to select the compounds with the best inhibition activity against SGLT2 stable cell line, the effect of samples on the ability of transport glucose were tested by flow cytometry. The interaction between fucus oligosaccharide and SGLT2 was analyzed by computer modeling. The proliferation model induced by high glucose was established in rat mesangial cells (HBZY-1 cells). The anti-DN activity of F2 was evaluated by MTT method, and the effect of optimal compounds on SphK1, NF-κB, p38 and ERK1/2 proteins in HBZY-1 cell model were investigated by Western Blot method. Results Five components were obtained after purification, and the optimal F2 component was determined by SGLT2 inhibitor screening, which has the characteristics of low molecular weight, high sulfate content and single monosaccharide composition. And 2-salfutated Fucose with eight degree of polymerization exhibit the strongest affinity interaction with SGLT2 in the research due to the effect of sulfate content and molecular weight of F2. The proliferation model of HBZY-1 cells was established successfully. It was found that F2 could inhibit mesangial proliferation at a low concentration, and its anti-mesangial proliferation effect was enhanced with the increase of concentration. And F2 exerted its anti-DN activity by inhibiting the expression of SphK1, NF-κB, p38 protein and phosphorylation of ERK1/2 protein. |
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