A stop-flow comprehensive two-dimensional HK-2 and HK-2/CIKI cell membrane chromatography comparative analysis system for screening the active ingredients from Pyrrosia calvata (Bak.) Ching against crystal-induced kidney injury
A stop-flow comprehensive two-dimensional HK-2 and HK-2/CIKI cell membrane chromatography comparative analysis system for screening the active ingredients from Pyrrosia calvata (Bak.) Ching against crystal-induced kidney injury
复制标题
停流综合二维HK-2和HK-2/CIKI细胞膜色谱比较分析系统筛选石韦抗晶体肾损伤活性成分
DOI：
10.1016/j.jpba.2020.113825
复制
发表时间：
2021-01-23
影响因子：
3.4
通讯作者：
Chen, Xiaofei
中科院分区：
文献类型：
--
作者：
Pan, Pengchao;Cheng, Jin;Chen, Xiaofei
晶体诱导的肾损伤（CIKI）是肾结石形成的基本病理变化，其分子机制尚不清楚。石韦Pyrrosia calvata（巴克.）清已被用于民间医药治疗尿石症多年。为进一步阐明石韦抗尿石症的药效物质及其作用机制，本研究建立了HK-2和HK-2/CIKI细胞膜色谱（CMC）二维对比分析系统，用于筛选石韦（巴克.）Ching对Ciki。二维CMC综合对比分析系统具有较好的选择性，对8种组分进行了筛选和鉴定。其中，芒果苷对HK-2/CIKI CMC柱的亲和力高于HK-2 CMC柱，并被选择用于进一步的有效性验证。细胞增殖实验表明芒果苷对草酸钠（NaOX）刺激的HK-2细胞具有保护作用。此外，在CIKI的啮齿动物模型中，芒果苷减少草酸钙（CaOX）晶体在小鼠肾脏中的沉积，减轻对肾组织的病理损伤，并抑制由CaOX晶体引起的OPN、MCP 1和CD 44表达的上调。所建立的二维CMC比较分析系统可用于中药中具有疾病特异性的活性成分的筛选，也适用于其他细胞模型。(C)2020爱思唯尔B. V.保留所有权利。
Crystal-induced kidney injury (CIKI) is the fundamental pathological change during nephrolithiasis, although the molecular mechanism is still unclear. Pyrrosia calvata (Bak.) Ching has been used in folk medicine to treat urolithiasis for years. To clarify the pharmacodynamic substances and the mechanism of its antiurolithiasis effects, in this study, a novel, stop-flow, comprehensive, two-dimensional (2D) HK-2 and HK-2/CIKI cell membrane chromatography (CMC) comparative analysis system was developed to screen for the potential active ingredients from Pyrrosia calvata (Bak.) Ching against CIKI. The comprehensive 2D CMC comparative analysis system showed satisfactory selectivity, and eight ingredients were screened and identified by this system. Among them, mangiferin exhibited higher affinity for the HK-2/CIKI CMC column than the HK-2 CMC column and was selected for further efficacy verification. Cell proliferation assays showed that mangiferin could protect HK-2 cell viability after stimulation with sodium oxalate (NaOX). Additionally, in a rodent model of CIKI, mangiferin decreased the deposition of calcium oxalate (CaOX) crystals in mouse kidneys, alleviated the pathological damage to kidney tissue, and inhibited the upregulation of OPN, MCP1, and CD44 expression caused by CaOX crystals. The established comprehensive 2D CMC comparative analysis system can be applied to screen active ingredients with disease specificity from traditional Chinese medicine (TCM) and is suitable for other cell models. (C) 2020 Elsevier B.V. All rights reserved.