对高糖损伤人脐静脉内皮细胞具有保护作用的天然植物提取物的筛选

摘要

目的 从五种药食同源植物(山药、枸杞、桑叶、银杏叶、桔梗)及丹参的提取物中筛选对高糖诱导的人脐静脉内皮细胞(HUVEC)具有保护作用的活性物质.方法 采用MTT方法检测分别由山药、枸杞、桑叶、银杏叶、桔梗、丹参提取物及盐酸曲美他嗪预处理的高糖诱导HUVEC细胞的存活率,初步筛选出对高糖诱导HUVEC细胞具有保护作用的活性物质及作用浓度.再采用微板法检测各组LDH、TNOS及iNOS的含量,硝酸还原酶法检测各组NO含量, ELISA法检测各组ET-1含量,苏木素伊红(HE)染色观察各组细胞形态学变化.结果 枸杞多糖(400 μg·mL-1)、桑叶黄酮(350 μg·mL-1)、银杏叶黄酮(125 μg·mL-1)对高糖诱导的HUVEC细胞活性保护作用明显,对抑制LDH、ET-1、iNOS和促进NO表达均有非常显著的效果(P<0.01);与空白对照组相比,细胞形态变化不大,排列紧密,呈不规则状,数量多,少数呈圆形.结论 枸杞多糖、桑叶黄酮及银杏叶黄酮对高糖诱导的人脐静脉内皮细胞损伤具有保护作用,这三种活性物质均来源于药食同源植物,具有安全可靠的特性,可以作为改善心血管疾病患者健康状况的保健类食品原料.%Objective To screen the active substances from the extracts of five edible medicinal plants (yam,Lycium barbarum,mulberry leaf,ginkgo leaf,Platycodon grandiflorum) and Salvia which have protective effect on high glucose-induced injury in human umbilical vein endothelial cells (HUVEC).Methods The viability of HUVEC was detected by MTT assay after treatment with yam extract,Lycium barbarum extract,mulberry leaf extract,ginkgo leaf extract,Platycodon grandiflorum extract or trimetazidine hydrochloride for the coarse screening of active substances.The levels of LDH,TNOS and iNOS were measured using micro plate method.The levels of NO and ET-1 were determined using nitrate reductase and ELISA,respectively.The morphological changes in HUVEC were observed by HE staining.Results Treatment with Lycium barbarum polysaccharide (400 μg·mL-1),mulberry leaf flavonoids (350 μg·mL-1) or Ginkgo leaf flavonoids (125 μg·mL-1) increased cell viability,reduced LDH,ET-1 and iNOS levels and promoted NO expression in high glucose-treated HUVEC (P<0.01).A large number of HUVEC were arranged densely with irregular shape,and a few cells were round in shape.Conclusion Lycium barbarum polysaccharide,mulberry leaf flavonoids and Ginkgo leaf flavonoids can protect against high glucose-induced injury in HUVEC.The three active substances are derived from edible medicinal plants and have safe and reliable characteristics.Therefore,they can be used as the healthy food ingredients for improving the health of patients with cardiovascular disease.