摘要
目的观察生地环烯醚萜苷类成分梓醇、毛蕊花糖苷对晚期糖基化终产物（AGEs）损伤人脐静脉内皮细胞（HUVEC）的保护作用。方法体外培养HUVEC,将细胞分为正常对照组、模型组、梓醇组及毛蕊花糖苷组,并设氨基胍组作为阳性对照。加入终浓度为0.05、0.1、0.5、1、5、10、100μmol/L的药物分别预孵1h后,加入200mg/L的AGEs作为刺激物继续培养,24h后用MTT法检测并计算药物对AGEs致HUVEC损伤的保护率。体外培养HUVEC,加入氨基胍及梓醇、毛蕊花糖苷（终浓度分别为1、10、100μmol/L）预孵1h后,加入终浓度为200mg/L的AGEs,另设模型组及空白对照组,24h后取各组细胞培养上清液,试剂盒测定超氧化物歧化酶（SOD）、丙二醛（MDA）、活性氧（ROS）和NO;ELISA法测定内皮素（ET）、血管紧张素Ⅱ（AngⅡ）、转化生长因子-（TGF-β）及白介素1（IL-1）。结果生地环烯醚萜苷类成分梓醇、毛蕊花糖苷（0.05、0.1、0.5、1、5、10、100μmol/L）可减轻AGEs致HUVEC的损伤。与空白对照组比较,模型组细胞上清液中ROS、AngⅡ、MDA含量增加（P〈0.05-0.01）,SOD活力和NO水平降低（P〈0.05-0.01）,TGF-β、IL-1β分泌增加（P〈0.05-0.01）。而梓醇、毛蕊花糖苷给药组（1、10、100μmol/L）能不同程度的降低ROS、AngⅡ、MDA含量,升高SOD活力和NO水平,减少TGF-、IL-1分泌,与模型组比较具有显著性差异（P〈0.05-0.01）。结论生地环烯醚萜苷类成分梓醇、毛蕊花糖苷可通过抑制氧化应激、调节内皮细胞分泌功能、抑制炎症因子及抗细胞粘连等保护血管内皮细胞。
OBJECTIVE To observe the protective effects of catalpol and verbascoside as habitat iridoid glycoside compo nents of Radix Rehmanniae respectively, on human umbilical vein endothelial cells （HUVEC） injured by advanced glycation end products （AGEs）. METHODS In vitro cultured HUVEC were divided into normal control group, model group, catalpol group and verbascoside group, while aminoguanidine group was set as a positive control. Each compound was diluted at con- centrations of 0.05, 0.1, 0.5, 1.5, 10, 100μmol/L and added into corresponding group. After pre-incubated for 1 h, 200 mg/ L of AGEs was added as a stimulus in all groups, except the normal control group. The HUVEC viabiIity was assessed by MTT treatment and the OD values were measured after 24 h incubation. To determine superoxide dismutase （SOD）, malondi- aldehyde （MDA）, reactive oxygen species （ROS） and NO in cell culture supernatant of each group by the corresponding kit, as well as to evaluate endothelin （ET）, angiotensin Ⅱ （AngⅡ）, transforming growth factor-β （TGF-β） and interleukin-1β （IL-1β） by ELISA, HUVEC were cultured in the same condition described above, with compounds treatment at concentrations of 1, 10 and 100μmol/L respectively. RESULTS Habitat iridoid glycoside components of Radix Rehmanniae, catalpol and ver- bascoside （0.05, 0.1, 0.5, 1.5, 10, 100 μmol/L）, could both attenuate the HUVEC injury induced by AGEs. Compared with the control group, the concentrations of ROS, Ang Ⅱ and MDA in the model group increased （P〈0.05-0.01）, the con- tents of SOD and NO decreased （P〈0.05-0.01）, and the secretion of TGF-β and IL-1β increased （P〈0.05-0.01）. To va tying degrees, catalpol and verbascoside （1, 10, 100 μmol/L） could decrease the concentrations of ROS, Ang Ⅱ and MDA, increase the levels of SOD and NO, and reduce the secretion of TGF-β and IL-1β, which showed a significant difference （P〈0. 05-0.01）, compared with the model group. CONCLUSION Catalpol and verbascoside as habitat iridoid glycoside compo- nents of Radix Rehmanniae can protect AGEs injured HUVEC by suppressing oxidative stress, regulate the secretion of endo- thelial cell and inhibit inflammatory cytokines and cell adhesion, etc.
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2015年第1期55-59,共5页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金(81073111)
