木豆叶、鲜地黄提取物对ox-LDL损伤的HUVEC的保护作用  

The Protective Effect of Cajanus Cajan Leaf Extract and Rehmannia Root Extract on ox-LDLinduced HUVEC Injury

作　　者：刘长河[1] 李开言[1] 张雪侠[1] 王艳艳[1] 李华妮 葛文静 刘方洲[1] 王晓丽[1] LIU Changhe;LI Kaiyan;ZHANG Xuexia;WANG Yanyan;LI Huani;GE Wenjing;LIU Fangzhou;WANGXiaoli(Henan Provincial Academy of Traditional Chinese Medicine and Pharmacy,Zhengzhou 450004 Henan,China)

机构地区：[1]河南省中医药研究院,河南郑州450004

出　　处：《中药新药与临床药理》2021年第8期1073-1078,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：河南省中医药科学研究专项(2015ZY02010、2016ZY2021、2017ZY2037、2018ZY2025);中国中医科学院中药研究所新药联合创制项目(2011xycz-04);郑州市金水区科技攻关项目(金科2014-33)。

摘　　要：目的研究木豆叶、鲜地黄提取物(DMT)对氧化低密度脂蛋白(ox-LDL)诱导损伤的人脐静脉内皮细胞(HUVEC)的影响,初步探讨DMT防治动脉粥样硬化的机制。方法应用血清药理学方法制备正常对照、辛伐他汀、DMT血清,以供细胞实验使用。采用ox-LDL损伤HUVEC,与各组药物共培养,即空白组、模型组、辛伐他汀组(15%辛伐他汀血清)及DMT低(5%DMT血清)、中(10%DMT血清)、高剂量组(15%DMT血清),24 h后MTT法检测细胞活力;收集细胞,流式细胞术检测细胞内活性氧(ROS)含量;硝酸还原酶法检测上清液NO含量;ELISA法检测上清液内皮素(ET-1)、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)含量;Western Blot法检测细胞凝集素样氧化低密度脂蛋白受体(Lox-1)、Caspase-3、Cytc蛋白的表达。结果与模型组比较,辛伐他汀组及DMT低、中、高剂量组细胞活力明显提高(P<0.01);DMT低剂量组ROS水平明显降低(P<0.05);辛伐他汀组细胞上清液ET-1、VCAM-1含量及Lox-1、Caspase-3、Cytc蛋白含量明显降低(P<0.05,P<0.01);DMT低剂量组细胞Caspase-3、Cytc蛋白含量明显降低(P<0.01);DMT中剂量组细胞上清液ET-1、VCAM-1含量及Lox-1、Caspase-3、Cytc蛋白含量明显降低(P<0.05,P<0.01),细胞上清液中NO含量明显升高(P<0.01);DMT高剂量组细胞上清液ET-1、VCAM-1含量及Lox-1、Cytc蛋白含量明显降低(P<0.01),NO含量明显升高(P<0.01)。结论DMT能够抑制ox-LDL诱导的HUVEC损伤,恢复其收缩舒张分泌功能,其机制可能与影响Lox-1介导的内皮细胞氧化应激有关。Objective To study the influence of extracts of Cajanus cajan leaf and Rehmannia root(DMT)on human umbilical vein endothelial cells(HUVEC),which were damaged by oxidized low-density lipoprotein(ox-LDL)and to preliminarily discuss the mechanism of prevention and treatment of atherosclerosis by DMT.Methods The serum containing the contrast,simvastatin and DMT was prepared and used in cell experiment.Ox-LDL was used to induce HUVEC injury and corresponding drugs were given for co-culture.Cultured HUVEC contained blank control group,model group,simvastatin group(15%simvastatin serum),DMT low dosage group(5%DMT serum),DMT medium dosage group(10%DMT serum),DMT high dosage group(15%DMT serum).The MTT method was used to detect the cell viability after 24 h culture.Flow cytometry was used to detect the reactive oxygen(ROS).The nitrate reductase activity method was used to detect the content of nitric oxide.The enzyme-linked immunosorbent assay(ELISA)was used to detect the content of endothelins(ET-1),intercellular cell adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in the supernatants.Western blot method was used to detect the protein expression of lectin-like oxidized low density lipoprotein receptor-1(Lox-1),Caspase-3 and Cytc.Results Compared with model group,the viability of HUVEC in 4 groups(simuvstatin group,DMT low dosage group,DMT medium dosage group and DMT high dosage group)was obviously improved(P<0.01).The level of ROS in DMT low dosage group was significantly decreased(P<0.05).The contents of ET-1,VCAM-1 and the expressions of Lox-1,Caspase-3 and Cytc protein in simuvstatin group were significantly reduced(P<0.05,P<0.01).The contents of Caspase-3、Cytc protein in DMT low dosage group were significantly reduced(P<0.01).The contents of ET-1,VCAM-1 and the expressions of Lox-1,Caspase-3 and Cytc protein in DMT medium dosage group were significantly reduced(P<0.05,P<0.01)and the content of nitric oxide in the cultured supernatant was significantly increased(P<0.01).The contents of ET-1�

关 键 词：木豆叶 鲜地黄 氧化低密度脂蛋白(ox-LDL) 人脐静脉内皮细胞(HUVEC) 细胞凝集素样氧化低密度脂蛋白受体(Lox-1) 氧化应激 

分 类 号：R285.5[医药卫生—中药学]