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
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摘要
目的观察两色金鸡菊黄酮类化合物黄诺玛苷对高糖诱导的人脐静脉内皮细胞(HUVEC)氧化应激及血管内皮生长因子(VEGF)和细胞间黏附分子-1(ICAM-1)蛋白表达的影响。方法将HUVEC细胞分为正常对照组(NG)、高糖模型组(HG)及不同浓度黄诺玛苷组(50、100、200、400μmol/L)。酶联免疫吸附测定法(ELASA)检测细胞上清液中丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)的活性。用Western blot方法检测细胞中VEGF和ICAM-1蛋白表达水平。结果与NG组比较,HG组MDA明显升高,SOD、CAT、GSH-Px明显降低(P<0.05或P<0.01)。与HG组比较,黄诺玛苷干预后,MDA含量显著降低(P<0.05),SOD、CAT、GSH-Px活力显著上升(P<0.05)。Western blot结果显示,与NG组比较,HG组VEGF和ICAM-1蛋白表达量增多(P<0.05或P<0.01)。与HG组比较,黄诺玛苷干预后VEGF和ICAM-1蛋白表达量随浓度递增而依次减少(P<0.05或P<0.01)。结论黄诺玛苷可能通过改善氧化应激水平和下调VEGF、ICAM-1的表达对高糖诱导HUVEC损伤具有一定保护作用。
Objective To observe the effects of flavanomarein glycosides of Coreopsis tinctoria Nutt. on oxidative stress and expression of vascular endothelial growth factor(VEGF) and intercellular cell adhesion molecule-1(ICAM-1) protein in human umbilical vein endothelial cells(HUVEC) induced by high glucose. Methods The cultured HUVEC cells were divided into normal control group(NG group), high glucose model group(HG group) and different concentrations of flavanomarein group(50, 100, 200, 400 μmol/L). The activity of malondialdehyde(MDA), superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px) in supernatant were detected by enzyme-linked immunosorbent assay(ELASA). The expression levels of VEGF and ICAM-1 protein in cells were detected by Western blot method.Results Compared with NG group, the content of MDA in HG group was significantly increased, the levels of SOD,CAT and GSH-Px in HG group were significantly decreased(P<0.05 or P<0.01). Compared with HG group, after flavanomarein intervention, the content of MDA in HG group was significantly decreased, CAT, GSH-Px activity increased significantly(P<0.05). The results of Western blot showed that compared with NG group, the expression of VEGF and ICAM-1 protein in HG group increased(P<0.05 or P<0.01). Compared with HG group, after flavanomarein intervention, the expression of VEGF and ICAM-1 protein increased with the increase of concentration followed by reduc tion(P<0.05 or P<0.01). Conclusion Flavanomarein has a protective effect on HUVEC induced by high glucose may be through improving the level of oxidative stress and decreasing the levels of VEGF and ICAM-1.
Objective To observe the effects of flavanomarein glycosides of Coreopsis tinctoria Nutt. on oxidative stress and expression of vascular endothelial growth factor(VEGF) and intercellular cell adhesion molecule-1(ICAM-1) protein in human umbilical vein endothelial cells(HUVEC) induced by high glucose. Methods The cultured HUVEC cells were divided into normal control group(NG group), high glucose model group(HG group) and different concentrations of flavanomarein group(50, 100, 200, 400 μmol/L). The activity of malondialdehyde(MDA), superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px) in supernatant were detected by enzyme-linked immunosorbent assay(ELASA). The expression levels of VEGF and ICAM-1 protein in cells were detected by Western blot method.Results Compared with NG group, the content of MDA in HG group was significantly increased, the levels of SOD,CAT and GSH-Px in HG group were significantly decreased(P<0.05 or P<0.01). Compared with HG group, after flavanomarein intervention, the content of MDA in HG group was significantly decreased, CAT, GSH-Px activity increased significantly(P<0.05). The results of Western blot showed that compared with NG group, the expression of VEGF and ICAM-1 protein in HG group increased(P<0.05 or P<0.01). Compared with HG group, after flavanomarein intervention, the expression of VEGF and ICAM-1 protein increased with the increase of concentration followed by reduc tion(P<0.05 or P<0.01). Conclusion Flavanomarein has a protective effect on HUVEC induced by high glucose may be through improving the level of oxidative stress and decreasing the levels of VEGF and ICAM-1.
引文
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[19]Fan WY,Liu NP.Meta-analysis of association between K469E polymorphism of the ICAM-1 gene and retinopathy in type 2 diabetes[J].Int J Ophthalmol,2015,8(3):603-607.
[20]Simoes 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.
[21]Rangasamy S,Mcguire PG,Das A.Diabetic retinopathy and inflammation:novel therapeutic targets[J].Middle East Afr J Ophthalmol,2012,19(1):52-59.
[22]Nakao S,Arima M,Ishikawa K,et al.Intravitreal antiVEGF therapy blocks inflammatory cell infiltration and re-entry into the circulation in retinal angiogenesis[J].Invest Ophth Vis Sci,2012,53(7):4323-4328.
[2]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.
[3]Liu M,Zhang Y,Liu X,et al.Protective effect ofα-melanocyte stimulating hormone on glutamate-induced retinal excitotoxicity[J].Zhonghua Shiyan Yanke Zazhi,2013,31(5):440-445.
[4]李冬明.昆仑雪菊的药学研究进展[J].浙江中医杂志,2012,47(10):776-777.
[5]古扎力努尔·艾尔肯,李新霞,毛新民等.新疆两色金鸡菊中绿原酸与总黄酮含量测定及指纹图谱的建立[J].西北药学杂志,2013,28(3):248-251.
[6]兰怡,卢伟,李琳琳,等.两色金鸡菊醇提物对实验性糖尿病大鼠血糖血脂的影响[J].中国药物应用与监测,2014,11(5):272-275.
[7]木合布力·阿布力孜,张燕,景兆均,等.新疆昆仑雪菊化学成分的初步定性研究[J].新疆医科大学学报,2010,33(6):628-630.
[8]卢伟,兰怡,李琳琳,等.两色金鸡菊醇提物对自发性高血压大鼠血压和血浆ET,AngⅡ,CGRP的影响[J].中国实验方剂学杂志,2014,20(5):149-153.
[9]张瑞,李新霞,毛新民,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(6):73-74.
[10]陈瑶,李新霞,李琳琳,等.两色金鸡菊血清药物化学研究[J].新疆医科大学学报,2017,40(4):420-424.
[11]李卉,张雨洁,姚蓝,等.两色金鸡菊乙酸乙酯提取物对糖尿病大鼠视网膜VEGF和ICAM1表达的影响[J].新疆医科大学学报,2017,40(4):411-414.
[12]Tryggestad JB,Vishwanath A,Jiang S,et al.Influence of gestational diabetes mellitus on human umbilical vein endothelial cell mi RNA[J].Clin Sci,2016,130(21):1955-1967.
[13]Krause BJ,Costello PM,Munozurrutia E,et al.Role of DNA methyltransferase 1 on the altered e NOS expression in human umbilical endothelium from intrauterine growth restricted fetuses[J].Clin Epigenetics,2013,8(9):944-952.
[14]Profumo E,Buttari B,D'Arcangelo D,et al.The Nutraceutical Dehydrozingerone and Its Dimer Counteract Inflammation-and Oxidative Stress-Induced Dysfunction of In Vitro Cultured Human Endothelial Cells:A Novel Perspective for the Prevention and The ra py of Atherosclerosis[J].Oxid Med Cell Longev,2016,2016:1246485.
[15]Adeshara KA,Diwan AG,Jagtap TR,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].Complicat,2017,31(2):439-448.
[16]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.
[17]Morita A,Ushikubo H,Mori A,et al.A delay in vascularization induces abnormal astrocyte proliferation and migration in the mouse retina[J].Dev Dyn,2017,246(3):186-200.
[18]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.
[19]Fan WY,Liu NP.Meta-analysis of association between K469E polymorphism of the ICAM-1 gene and retinopathy in type 2 diabetes[J].Int J Ophthalmol,2015,8(3):603-607.
[20]Simoes 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.
[21]Rangasamy S,Mcguire PG,Das A.Diabetic retinopathy and inflammation:novel therapeutic targets[J].Middle East Afr J Ophthalmol,2012,19(1):52-59.
[22]Nakao S,Arima M,Ishikawa K,et al.Intravitreal antiVEGF therapy blocks inflammatory cell infiltration and re-entry into the circulation in retinal angiogenesis[J].Invest Ophth Vis Sci,2012,53(7):4323-4328.