朱日很滴丸含药血清对ox-LDL损伤的HUVEC转录因子Nrf2核转位的影响  

Effect of Serum Containing Zhurihen Dropping Pill on the Nuclear Translocation of Transcription Factor Nrf2 in HUVEC Damaged by ox-LDL

作　　者：王跃武[1] 韩瑞兰 郭俊英 邢煜舒 Wang Yuewu;Han Ruilan;Guo Junying;Xing Yushu(Engineering Research Center for New Drug Screening,Inner Mongolia Medical University,Inner Mongolia Autonomous Region,Hohhot 010110;Department of Pharmacology,Inner Mongolia Medical University,Inner Mongolia Autonomous Region,Hohhot 010110)

机构地区：[1]内蒙古医科大学新药筛选工程研究中心,呼和浩特010110 [2]内蒙古医科大学药学院,呼和浩特010110

出　　处：《中药药理与临床》2019年第5期121-125,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：内蒙古自然科学基金(2017MS08123);内蒙古自治区高等教育基金项目(NZRY17118)

摘　　要：目的:研究朱日很滴丸含药血清对经ox-LDL损伤的HUVEC的保护作用,探讨其可能的作用机制。方法:将氧化低密度脂蛋白(ox-LDL)与人脐静脉内皮细胞(HUVEC)共孵育,建立内皮细胞损伤模型,加入朱日很滴丸含药血清进行干预。MTS法检测朱日很滴丸含药血清对氧化损伤内皮细胞的活性影响;实时荧光定量PCR法(qPCR)和蛋白免疫印迹法(Westorn blot)检测朱日很滴丸含药血清对内皮细胞氧化损伤模型中Nrf2的mRNA水平和蛋白表达的影响;细胞免疫荧光法检测HUVEC中Nrf2蛋白的核转位情况。结果:与空白对照组比较,模型组、溶媒对照血清组细胞存活率明显降低,Nrf2 mRNA及蛋白表达明显下调,核质比明显降低。与溶媒对照血清组相比,朱日很滴丸各剂量20%含药血清组可提高氧化损伤的内皮细胞的活性,Nrf2的mRNA水平和蛋白表达明显上调,朱日很滴丸0.79 g/kg组20%含药血清,Nrf2蛋白的核、质比明显增加;20%含药血清各剂量组细胞核中Nrf2荧光强度明显增强,且有剂量依赖性。结论:朱日很滴丸可明显降低HUVEC的氧化应激损伤,其作用机制可能与促进转录因子Nrf2核转位有关。Objective:To study the protective effect of serum containing Zhurihen Dropping Pills on HUVEC(human umbilical vein endothelial cells)injured by ox-LDL and explore its possible mechanism.Methods:Oxidative low density lipoprotein(ox-LDL)was co-incubated with HUVEC to establish the endothelial cell injury model.Serum containing Zhurihen dropping pills was added to intervene the model.MTS method was used to detect the activity of endothelial cells by oxidative damage.Real-time quantitative PCR(qPCR)and Western blot method were used to observe the levels of protein expressions and mRNA of Nrf2.Cellular immunofluorescence assay was used to detect the nuclear translocation of Nrf2 protein in HUVEC.Results:Compared with the control group,cell viability was decreased,the levels of protein expressions and mRNA of Nrf2 were significantly down regulated,the nucleo-cytoplasmic ratio was significantly decreased in the model group.Compared with the model group,20%serum containing Zhurihen Dropping Pills in all doses of pills groups increased the activity of endothelial cells damaged by oxidative stress.The levels of Nrf2 mRNA and protein in drug-containing serum group was significantly increased.Compared with the model group,the nucleo-cytoplasmic ratio was significantly increased in 0.79 g/kg serum containing Zhurihen Dropping Pills group.The fluorescence intensity of Nrf2 in the nuclei was enhanced in all drug groups,in dose dependent manner.Conclusion:Zhurihen Dropping Pills can significantly alleviate the oxidative stress injury of HUVEC,its mechanism of antioxidant damage may be related to the promotion of nuclear translocation of transcription factor Nrf2.

关 键 词：朱日很滴丸 OX-LDL HUVEC NRF2 

分 类 号：R28[医药卫生—中药学]