摘要
晚期糖基化终产物(advanced glycation end products,AGEs)是食品在热加工和贮藏过程中形成的一类化合物,与心血管疾病、糖尿病、肾病等多种慢性病的发生有重要联系。本实验研究了含有AGEs的面包皮水提取物(bread crust extract,BCE)对人肾小管上皮细胞(human kidney tubular epithelial cells,HKCs)内氧化应激的影响及细胞损伤。采用不同质量浓度的BCE处理HKCs,并经噻唑蓝法确定后续BCE处理质量浓度;通过检测细胞内活性氧(reactive oxygen species,ROS)、总超氧化物歧化酶(superoxide dismutase,SOD)、脂质过氧化产物丙二醛(malondialdehyde,MDA)及培养上清液中乳酸脱氢酶(lactic dehydrogenase,LDH)水平,评价BCE对HKCs胞内氧化应激平衡及细胞损伤程度的影响。结果表明:不同质量浓度的BCE处理24 h后,HKCs胞内ROS水平随着BCE质量浓度的增大不断升高,且在8 mg/m L和12 mg/m L时胞内ROS水平达到最大值;与对照组相比,胞内MDA含量在BCE质量浓度为4 mg/m L和8 mg/m L时极显著增加(P<0.01),但BCE质量浓度达12 mg/m L时,MDA含量却显著降低(P<0.05)。相反,胞内总SOD活力却随着BCE质量浓度的增大而降低,在8 mg/m L和12 mg/m L时,胞内总SOD活力发生极显著降低(P<0.01)。胞外分泌型LDH水平随着BCE处理质量浓度的增大而升高,在BCE质量浓度为8 mg/m L和12 mg/m L时,LDH活力发生极显著增加(P<0.01)。因此,BCE可以通过提高HKCs胞内活性氧水平,引起胞内总SOD活力降低,进而促进细胞发生脂质过氧化反应,改变细胞膜通透性,使胞外LDH活力升高,导致细胞严重受损。
Advanced glycation end products （AGEs） are a class of compounds that are formed during thermal processing and storage of foods, which are signifcantly associated with various chronic diseases, such as cardiovascular diseases, diabetes, and kidney diseases. In the present study, bread crust extract （BCE）, rich in AGEs, was obtained from bread crust through water extraction, and we investigated if BCE could cause intracellular oxidative stress and cell damage in human kidney tubular epithelial cells （HKCs）. HKCs were treated with different BCE concentrations for determining the appropriate concentration of BCE by 3-（4,5）-dimethylthiahiazo （-z-y1）-3,5-di-phenytetrazoliumromide assay for use in subsequent experiments. The effect of BCE on intracellular oxidative stress balance and the degree of cell damage was evaluated by detecting the levels of intracellular reactive oxygen species （ROS）, total superoxide dismutase （SOD） and malondialdehyde （MDA） and lactic dehydrogenase （LDH） in supernatants. The results indicated that the level of ROS in HKCs was increased along with increasing BCE concentration after being cultured with different concentrations of BCE for 24 h, and reached a maximum value at 8 and 12 mg/mL. Intracellular MDA contents increased signifcantly at 4 and 8 mg/mL of BCE （P 〈 0.01）, but markedly declined at 12 mg/mL （P 〈 0.05）. On the contrary, intracellular SOD activity was reduced with increasing concentration of BCE, and signifcantly decreased at BCE concentrations of 8 and 12 mg/mL. The level of secreted LDH was increased with increasing BCE concentrations, and the increase was highly signifcant at 8 and 12 mg/mL （P 〈 0.01）.Therefore, BCE can cause serious damage to HKCs via elevating intracellular ROS levels, inhibiting total SOD activity and thereby promoting lipid peroxidation reaction, altering the cell membrane permeability, and consequently improving extracellular LDH activity.
作者
贾本盼
袁晓金
范智义
胡静
李巨秀
JIA Benpan;YUAN Xiaojin;FAN Zhiyi;HU Jing;LI Juxiu(College of Food Science and Engineering, Northwest A ＆ F University, Yangling 712100, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第1期136-141,共6页
Food Science
基金
国家自然科学基金面上项目(31471579)
关键词
人肾小管上皮细胞
面包皮水提取物
氧化损伤
脂质过氧化
human kidney tubular epithelial cells
bread crust extract
oxidative damage
lipid peroxidation
