马里苷对高糖损伤人脐静脉内皮细胞的作用
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摘要
目的观察两色金鸡菊黄酮类化合物马里苷对高糖损伤人脐静脉内皮细胞(HUVEC)的作用。方法取对数生长期的细胞,分为正常对照组(NG)、高糖模型组(HG)及马里苷不同浓度(50、100、200、400μmol/L)给药组。采用CCK-8法检测细胞活力;ELASA法检测MDA含量及SOD、CAT、GSH-PX活力; Western blot法检测炎性因子VEGF、ICAM-1、MCP-1在HUVEC细胞中的蛋白表达。结果与NG组比较,及HG组细胞活力明显降低(P<0.05); MDA含量显著升高(P<0.05),SOD、CAT、GSH-PX活性降低(P<0.05);与HG组(50 mmol/L)比较,马里苷药物干预组(50、100、200、400μmol/L)细胞活性明显增加,差异有统计学意义(P<0.05),且随浓度增加细胞活性明显增加。Western blot蛋白印迹结果显示,与NG组比较,HG组VEGF、ICAM-1、MCP-1蛋白表达明显增加,差异有统计学意义(P<0.05);与HG组比较,马里苷干预组VEGF、ICAM-1、MCP-1蛋白表达显著降低,差异有统计学意义(P<0.05)。结论马里苷对高糖损伤的HUVEC细胞具有一定保护作用。
Objective To observe the effects of marein,a flavonoid compound in plains coreopsis,on high glucose induced damage in human umbilical vein endothelial cells( HUVEC). Methods The cells in logarithmic growth phase were taken and divided into the normal group,high glucose model group and marein groups with different concentrations( 50,100,200,400 μmol/L). The cell viability was detected by CCK-8. The content of malondialdehyde( MDA),the activities of superoxide dismutase( SOD),catalase( CAT) and glutathione-peroxidase( GSH-PX) were assessed by enzyme-linked immunosorbent assay( ELISA),the protein expressions of vascular endothelial factor( VEGF),intercellular adhesion molecule-1( ICAM-1) and monocyte chemoattractant protein-1( MCP-1) in HUVEC were detected by Western blot. Results Compared with the normal group,the cell viability was significantly decreased in high glucose model groups( P<0.05),while the MDA content was significantly increased( P<0.05) and the activities of SOD,CAT and GSH-PX were decreased( P<0.05).Compared with the high glucose model group( 50 mmol/L),the cell viability was obviously increased in marein treatment groups( 50,100,200 and400 μmol/L),with statistically significant differences( P<0.05),and the cell viability was significantly increased with the increasing concentration of marein. The results of Western blot showed that the protein expressions of VEGF,ICAM-1 and MCP-1 were increased significantly in the high glucose group compared with the normal control group,with statistically significant differences( P<0.05). Compared with the model group,the protein expressions of VEGF,ICAM-1 and MCP-1 were significantly decreased in the marein treatment groups,with statistically significant differences( P<0.05). Conclusion Marein exerts a certain protective effect on high glucose-induced HUVEC damage.
Objective To observe the effects of marein,a flavonoid compound in plains coreopsis,on high glucose induced damage in human umbilical vein endothelial cells( HUVEC). Methods The cells in logarithmic growth phase were taken and divided into the normal group,high glucose model group and marein groups with different concentrations( 50,100,200,400 μmol/L). The cell viability was detected by CCK-8. The content of malondialdehyde( MDA),the activities of superoxide dismutase( SOD),catalase( CAT) and glutathione-peroxidase( GSH-PX) were assessed by enzyme-linked immunosorbent assay( ELISA),the protein expressions of vascular endothelial factor( VEGF),intercellular adhesion molecule-1( ICAM-1) and monocyte chemoattractant protein-1( MCP-1) in HUVEC were detected by Western blot. Results Compared with the normal group,the cell viability was significantly decreased in high glucose model groups( P<0.05),while the MDA content was significantly increased( P<0.05) and the activities of SOD,CAT and GSH-PX were decreased( P<0.05).Compared with the high glucose model group( 50 mmol/L),the cell viability was obviously increased in marein treatment groups( 50,100,200 and400 μmol/L),with statistically significant differences( P<0.05),and the cell viability was significantly increased with the increasing concentration of marein. The results of Western blot showed that the protein expressions of VEGF,ICAM-1 and MCP-1 were increased significantly in the high glucose group compared with the normal control group,with statistically significant differences( P<0.05). Compared with the model group,the protein expressions of VEGF,ICAM-1 and MCP-1 were significantly decreased in the marein treatment groups,with statistically significant differences( P<0.05). Conclusion Marein exerts a certain protective effect on high glucose-induced HUVEC damage.
引文
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[4]LIU B,LI C P,WANG W Q,et al.Lignans extracted from Eucommia Ulmoides Oliv.protects against Ages-induced retinal endothelial cell injury[J].Cell Physiol Biochem,2016,39(5):2044-2054.
[5]于晓忠,李金涛,刘志广.血清胆红素水平与2型糖尿病视网膜病变的关系[J].贵阳中医学院学报,2013,35(6):171-172.
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[9]刘伟新,邓继华,徐鸿.一种金鸡菊花的生药学研究[J].中国民族医药杂志,2009,15(1):24-25.
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[11]李卉.新疆两色金鸡菊对糖尿病视网膜病变的影响及分子机理研究[D].乌鲁木齐:新疆医科大学,2017.
[12]于晓忠,李金涛,刘志广.血清胆红素水平与2型糖尿病视网膜病变的关系[J].贵阳中医学院学报,2013,35(6):171-172.
[13]张瑞,李新霞,李琳琳,等.HPLC法测定两色金鸡菊提取物中Flavanomarein和Marein的含量[J].安徽农业科学,2015,43(6):73-74,128.
[14]古扎力努尔·艾尔肯,王健,兰怡,等.两色金鸡菊花提取物的抗氧化活性测定及TLC-Bio分析抗氧化活性成分[J].新疆医科大学学报,2016,39(7):858-861.
[15]高飞.雪菊总黄酮降血脂作用及其机制研究[D].苏州:苏州大学,2016.
[16]刘菁,姚蓝,毕晓娟,等.两色金鸡菊黄酮成分马里苷对胰岛β细胞的保护作用[J].新疆医科大学学报,2017,40(4):428-432.
[17]TRYGGESTAD J B,VISHWANATH A,JIANG S,et al.Influence of gestational diabetes mellitus on hμMan u Mbilical vein endothelial cell mi RNA[J].Clin Sci,2016,130(21):1955-1967.
[18]KRAUSE B J,COSTELLO P M,MUOZURRUTIA E,et al.Role of DNA methyltransferase 1 on the altered e NOS expression in hμManμMbilical endotheliμM from intrauterine growth restricted fetuses[J].Clin Epigenetics,2013,8(9):944-952.
[19]PROFUMO E,BUTTARI B,D’ARCANGELO D,et al.The nutraceutical dehydrozingerone and its dimer counteract inflammationand oxidative stress-induced dysfunction ofin vitrocultured human endothelial cells:a novel perspective for the prevention and therapy of atherosclerosis[J].Oxid Med Cell Longev,2016(3):1-12.
[20]ADESHARA K A,DIWAN A G,JAGTAP T R,et al.Relationship between plasma glycation with membrane modification,oxidative stress and expression of glucose trasporter-1 in type 2 diabetes patients with vascular complications[J].J Diabetes Complicat,2017,31(2):439-448.
[21]ARDEN GEOFFREY B,SIVAPRASAD SOBHA.Hypoxia and oxidative stress in the causation of diabetic retinopathy[J].Curr Diabetes Rev,2011,7(5):291-304.
[22]王莉莉,刘堃,许迅,等.糖尿病性视网膜病变抗血管内皮生长因子治疗进展[J].上海交通大学学报(医学版),2012,32(2):219-225.
[23]MORITA A,USHIKUBO H,MORI A,et al.A delay in vascularization induces abnormal astrocyte proliferation and migration in the mouse retina[J].Dev Dynam,2017,246(3):186-200.
[24]AMATO R,BIAGIONI M,CAMMALLERI M,et al.VEGF as a survival factor in ex vivo models of early diabetic retinopathy[J].Invest Ophthalmol Vis Sci,2016,57(7):3066-3076.
[25]FAN W Y,LIU N P.Meta-analysis of association between K469Epolymorphism of the ICAM-1 gene and retinopathy in type 2 diabetes[J].Int J Ophthalmol,2015,8(3):603-607.
[26]SIMES MJ,LOBO C,EGAS C,et al.Genetic variants in ICAM1,PPARGC1A and MTHFR are potentially associated with different phenotypes of diabetic retinopathy[J].Acta Ophthalmol,2014,232(3):156-162.
[27]CAPEANS C,DE ROJAS M V,LOJO S,et al.C-C chemokines in the vitreous of patients with proliferative vitreoretinopathy and proliferative diabetic retinopathy[J].Retina,1998,18(6):546-550.
[28]MIGDAL C,BOTTON J,EL A Z,et al.Reactivity of chemical sensitizers toward amino acids in cellulo plays a role in the activation of the Nrf2-ARE pathway in human monocyte dendritic cells and the THP-1 cell line[J].Toxicol Sci,2013,133(2):259.
[29]陈慷,胡世兴,邓新国,等.单核细胞趋化蛋白-1在早期糖尿病大鼠视网膜中的表达及意义[J].中华实验眼科杂志,2005,23(1):23-25.
[30]LIBBY P,SUKHOVA G,LEE R T,et al.Cytokines regulate vascular functions related to stability of the atherosclerotic plaque[J].JCardiovasc Pharmacol,1995,25(Suppl 2):S9-S12.
[31]SOZZANI S,LOCATI M,ZHOU D,et al.Receptors,signal transduction,and spectrum of action of monocyte chemotactic protein-1 and related chemokines[J].J Leukocyte Biol,1995,57(5):788-794.
[32]RANGASAMY S,MCGUIRE P G,DAS A.Diabetic retinopathy and inflammation:novel therapeutic targets[J].Meago,2012,19(1):52-59.
[33]NAKAO S,ARIMA M,ISHIKAWA K,et al.Intravitreal anti-VEGFtherapy blocks inflammatory cell infiltration and re-entry into the circulation in retinal angiogenesis[J].Invest Ophth Vis Sc,2012,53(7):4323-4328.
[34]陈永钧,龙晓英,潘素静,等.黄酮类化合物的药效机制及构效关系研究进展[J].中国实验方剂学杂志,2013,19(11):337-344.
[35]张红雨.黄酮类抗氧化剂结构活性关系的理论解释[J].中国科学,1999,29(1):91-96.
[36]王丽丽,李琳琳,张楠楠,等.黄诺玛苷对高糖诱导HUVEC细胞氧化应激及VEGF和ICAM-1蛋白表达的影响[J].中国医药导报,2018,15(10):8-12.
[2]CHEUNG N,TIKELLIS G,WANG J J.Diabetic retinopathy[J].New Engl J Med,2010,114(11):2098.
[3]YAU J W Y,ROGERS S L,KAWASAKI R,et al.Global Prevalence and major risk factors of diabetic retinopathy[J].Diabetes Care,2016,35(3):556-564.
[4]LIU B,LI C P,WANG W Q,et al.Lignans extracted from Eucommia Ulmoides Oliv.protects against Ages-induced retinal endothelial cell injury[J].Cell Physiol Biochem,2016,39(5):2044-2054.
[5]于晓忠,李金涛,刘志广.血清胆红素水平与2型糖尿病视网膜病变的关系[J].贵阳中医学院学报,2013,35(6):171-172.
[6]ZHANG L,DONG L,LIU X,et al.α-Melanocyte-stimulating hormone protects retinal vascular endothelial cells from oxidative stress and apoptosis in a rat model of diabetes[J].PLoS One,2014,9(4):e93433.
[7]CHERNYKH V V,VARVARINSKY E V,SMIRNOV E V,et al.Proliferative and inflammatory factors in the vitreous of patients with proliferative diabetic retinopathy[J].Indian J Ophthalmol,2015,63(1):33-36.
[8]WANG Y,SUBRAMANIAN P,SHEN D,et al.Pigment epitheliumderived factor reduces apoptosis and pro-inflammatory cytokine gene expression in a murine model of focal retinal degeneration[J].Asn Neuro,2013,5(5):3522-3528.
[9]刘伟新,邓继华,徐鸿.一种金鸡菊花的生药学研究[J].中国民族医药杂志,2009,15(1):24-25.
[10]姜保平.两色金鸡菊茶饮对2型糖尿病胰岛素抵抗的预防作用及机制研究[D].北京:北京协和医学院,2015.
[11]李卉.新疆两色金鸡菊对糖尿病视网膜病变的影响及分子机理研究[D].乌鲁木齐:新疆医科大学,2017.
[12]于晓忠,李金涛,刘志广.血清胆红素水平与2型糖尿病视网膜病变的关系[J].贵阳中医学院学报,2013,35(6):171-172.
[13]张瑞,李新霞,李琳琳,等.HPLC法测定两色金鸡菊提取物中Flavanomarein和Marein的含量[J].安徽农业科学,2015,43(6):73-74,128.
[14]古扎力努尔·艾尔肯,王健,兰怡,等.两色金鸡菊花提取物的抗氧化活性测定及TLC-Bio分析抗氧化活性成分[J].新疆医科大学学报,2016,39(7):858-861.
[15]高飞.雪菊总黄酮降血脂作用及其机制研究[D].苏州:苏州大学,2016.
[16]刘菁,姚蓝,毕晓娟,等.两色金鸡菊黄酮成分马里苷对胰岛β细胞的保护作用[J].新疆医科大学学报,2017,40(4):428-432.
[17]TRYGGESTAD J B,VISHWANATH A,JIANG S,et al.Influence of gestational diabetes mellitus on hμMan u Mbilical vein endothelial cell mi RNA[J].Clin Sci,2016,130(21):1955-1967.
[18]KRAUSE B J,COSTELLO P M,MUOZURRUTIA E,et al.Role of DNA methyltransferase 1 on the altered e NOS expression in hμManμMbilical endotheliμM from intrauterine growth restricted fetuses[J].Clin Epigenetics,2013,8(9):944-952.
[19]PROFUMO E,BUTTARI B,D’ARCANGELO D,et al.The nutraceutical dehydrozingerone and its dimer counteract inflammationand oxidative stress-induced dysfunction ofin vitrocultured human endothelial cells:a novel perspective for the prevention and therapy of atherosclerosis[J].Oxid Med Cell Longev,2016(3):1-12.
[20]ADESHARA K A,DIWAN A G,JAGTAP T R,et al.Relationship between plasma glycation with membrane modification,oxidative stress and expression of glucose trasporter-1 in type 2 diabetes patients with vascular complications[J].J Diabetes Complicat,2017,31(2):439-448.
[21]ARDEN GEOFFREY B,SIVAPRASAD SOBHA.Hypoxia and oxidative stress in the causation of diabetic retinopathy[J].Curr Diabetes Rev,2011,7(5):291-304.
[22]王莉莉,刘堃,许迅,等.糖尿病性视网膜病变抗血管内皮生长因子治疗进展[J].上海交通大学学报(医学版),2012,32(2):219-225.
[23]MORITA A,USHIKUBO H,MORI A,et al.A delay in vascularization induces abnormal astrocyte proliferation and migration in the mouse retina[J].Dev Dynam,2017,246(3):186-200.
[24]AMATO R,BIAGIONI M,CAMMALLERI M,et al.VEGF as a survival factor in ex vivo models of early diabetic retinopathy[J].Invest Ophthalmol Vis Sci,2016,57(7):3066-3076.
[25]FAN W Y,LIU N P.Meta-analysis of association between K469Epolymorphism of the ICAM-1 gene and retinopathy in type 2 diabetes[J].Int J Ophthalmol,2015,8(3):603-607.
[26]SIMES MJ,LOBO C,EGAS C,et al.Genetic variants in ICAM1,PPARGC1A and MTHFR are potentially associated with different phenotypes of diabetic retinopathy[J].Acta Ophthalmol,2014,232(3):156-162.
[27]CAPEANS C,DE ROJAS M V,LOJO S,et al.C-C chemokines in the vitreous of patients with proliferative vitreoretinopathy and proliferative diabetic retinopathy[J].Retina,1998,18(6):546-550.
[28]MIGDAL C,BOTTON J,EL A Z,et al.Reactivity of chemical sensitizers toward amino acids in cellulo plays a role in the activation of the Nrf2-ARE pathway in human monocyte dendritic cells and the THP-1 cell line[J].Toxicol Sci,2013,133(2):259.
[29]陈慷,胡世兴,邓新国,等.单核细胞趋化蛋白-1在早期糖尿病大鼠视网膜中的表达及意义[J].中华实验眼科杂志,2005,23(1):23-25.
[30]LIBBY P,SUKHOVA G,LEE R T,et al.Cytokines regulate vascular functions related to stability of the atherosclerotic plaque[J].JCardiovasc Pharmacol,1995,25(Suppl 2):S9-S12.
[31]SOZZANI S,LOCATI M,ZHOU D,et al.Receptors,signal transduction,and spectrum of action of monocyte chemotactic protein-1 and related chemokines[J].J Leukocyte Biol,1995,57(5):788-794.
[32]RANGASAMY S,MCGUIRE P G,DAS A.Diabetic retinopathy and inflammation:novel therapeutic targets[J].Meago,2012,19(1):52-59.
[33]NAKAO S,ARIMA M,ISHIKAWA K,et al.Intravitreal anti-VEGFtherapy blocks inflammatory cell infiltration and re-entry into the circulation in retinal angiogenesis[J].Invest Ophth Vis Sc,2012,53(7):4323-4328.
[34]陈永钧,龙晓英,潘素静,等.黄酮类化合物的药效机制及构效关系研究进展[J].中国实验方剂学杂志,2013,19(11):337-344.
[35]张红雨.黄酮类抗氧化剂结构活性关系的理论解释[J].中国科学,1999,29(1):91-96.
[36]王丽丽,李琳琳,张楠楠,等.黄诺玛苷对高糖诱导HUVEC细胞氧化应激及VEGF和ICAM-1蛋白表达的影响[J].中国医药导报,2018,15(10):8-12.