摘要
探讨了山楂果胶低聚半乳糖醛酸提取物对中波紫外线(ultraviolet B,UVB)辐射后人永生化表皮角质形成细胞(HaCaT)的氧化损伤和光老化的保护作用。体外培养HaCaT细胞,分为对照组(HaCaT细胞不照射UVB,不给予山楂果胶低聚半乳糖醛酸提取物处理)、UVB辐射组(30 mJ/cm^2)、低剂量(5μg/mL)山楂果胶低聚半乳糖醛酸+30 mJ/cm^2 UVB辐射组、高剂量(10μg/mL)山楂果胶低聚半乳糖醛酸+30 mJ/cm^2 UVB辐射组。以噻唑蓝比色法检测细胞活力,采用试剂盒检测超氧化物歧化酶(superoxide dismulase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)和乳酸脱氢酶(lactate dehydrogenase,LDH)活力,丙二醛(malondialdehyde,MDA)含量,羟脯氨酸(hydroxyproline,Hyp)质量浓度和活性氧(reactive oxygen species,ROS)水平。采用酶联免疫吸附测定实验检测肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白细胞介素-6(interleukin-6,IL-6)质量浓度和基质金属蛋白酶-1(matrix metalloproteinase 1,MMP-1)含量。结果表明,经UVB照射后,Ha Ca T细胞活力明显降低。低、高剂量的山楂果胶低聚半乳糖醛酸提取物对经UVB辐射的HaCaT细胞具有显著的增殖作用(P<0.05);SOD、GSH-Px、CAT活力增加;LDH活力、ROS水平和MDA含量下降;Hyp质量浓度增加;MMP-1含量和TNF-α、IL-6质量浓度下降,且与辐射组比较均具有显著或极显著差异(P<0.05或P<0.01)。实时荧光定量聚合酶链式反应检测MMP-1、TNF-α及IL-6 m RNA表达水平与其含量或质量浓度检测结果一致。因此,山楂果胶低聚半乳糖醛酸对UVB辐射Ha Ca T细胞的氧化损伤及光老化具有保护作用。
n this study,the protective effects of hawthorn pectin oligogalacturonide extract against oxidative damage and photoaging induced by ultraviolet B(UVB)in HaCaT cells were explored.HaCaT cells were cultured and divided into four groups,including control group,radiation(30 mJ/cm2)group,low-dose(5μg/mL oligogalacturonide extract)with 30 mJ/cm2 radiation group,and high-dose(10μg/mL oligogalacturonide extract)with 30 mJ/cm2 radiation group.Cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium(MTT)method.The activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT)and lactate dehydrogenase(LDH)were detected using commercial kits,and the contents of malondialdehyde(MDA),hydroxyproline(Hyp)and reactive oxygen species(ROS)were also measured.The secretion levels of tumor necrosis factor alpha(TNF-α),interleukin-6(IL-6)and matrix metalloproteinase 1(MMP-1)were evaluated using enzyme-linked immunosorbent assay.The results showed that the cell viability of HaCaT cells significantly decreased after UVB exposure.Low-and high-dose hawthrown pectin oligogalacturonide extract had a significant growth promoting effect on UVB-irradiated HaCaT cells(P<0.05).In addition,the extract significantly increased the activities of SOD,GSH-Px,and CAT,decreased LDH activity,ROS level and MDA content,augmented hydroxyproline concentration,and reduced the secretion levels of MMP-1,TNF-αand IL-6 compared with the model group.The mRNA expression levels of MMP-1,TNF-αand IL-6 as determined by realtime polymerase chain reaction were in agreement with the production of MMP-1,TNF-α,and IL-6.Therefore,hawthrown pectin oligogalacturonide has a protective effect against UVB radiation-induced oxidative damage and photoaging in HaCaT cells.
作者
刘素稳
吴瞻邑
由璐
常学东
LIU Suwen;WU Zhanyi;YOU Lu;CHANG Xuedong(College of Food Science and Technology,Hebei Normal University of Science and Technology,Qinhuangdao 066604,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第21期210-218,共9页
Food Science
基金
河北省科技计划项目(16237113D)
秦皇岛市科技计划项目(201402B022)
