华夏小葱制剂对氧化低密度脂蛋白诱导内皮细胞损伤的影响及相关机制  

Protective effect of Huaxia shallot preparation on human umbilical vein endothelial cell injury induced by oxidized low density lipoprotein and its mechanism

作　　者：张介眉[1] 郭洁[1] 涂欣[2] 时昭红[1] 郝建军[1] 柯于鹤[1] 关江峰[1] 何娟娟[1] 

机构地区：[1]武汉市第一医院中西医结合心血管疾病研究室,湖北武汉430022 [2]华中科技大学生命科学与技术学院人类基因组研究中心,湖北武汉430074

出　　处：《中西医结合学报》2007年第6期675-680,共6页Journal of Chinese Integrative Medicine

基　　金：湖北省自然科学基金(No.2006ABA327)

摘　　要：目的:观察人脐静脉内皮细胞在氧化型低密度脂蛋白(oxidized low density lipoprotein,Ox-LDL)损伤作用下,其细胞形态、活力和凋亡率的改变及华夏小葱制剂的干预作用,并探讨其可能的分子作用靶点。方法:体外培养脐静脉内皮细胞,制备Ox-LDL,将100μg/L Ox-LDL和不同药物分别作用于内皮细胞,24h后观察各组的细胞形态,显微镜下细胞计数,甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法测定各组和空白对照组的细胞活力以及细胞上清液中一氧化氮(nitric oxide,NO)含量和超氧化物歧化酶(superoxide dismutase,SOD)的活性,用逆转录-多聚酶链反应(reverse transcription polymerase chain reac- tion,RT-PCR)技术结合图像分析技术观察过氧化物酶体增殖激活受体γ(peroxisome proliferators activa- ted receptorγ,PPARγ)及内支型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)表达的变化,用蛋白免疫印迹(Western blot)技术测定细胞PPAR-γ及eNOS蛋白的表达。结果:Ox-LDL能抑制内皮细胞分泌NO的活性并增加内皮细胞的凋亡,与空白对照组比较差异有统计学意义(P<0.05)。华夏小葱制剂能通过上调PPAR-γ及eNOS的表达来刺激内皮细胞分泌NO的活性,同时减少内皮细胞的凋亡,与Ox-LDL组比较差异有统计学意义(P<0.05),其效果仅次于平福注射液,优于丹参滴丸及立普妥。结论:华夏小葱制剂使内皮细胞NO活性增强的同时降低了细胞凋亡率,其保护内皮细胞的作用机制可能与调节PPAR-γ及eNOS表达有关。Objective： To observe the protective effect of Huaxia cell （HUVEC） injury induced by oxidized low density shallot preparation on human umbilical vein endothelia poprotein （Ox-LDL） in vitro. Methods： Ox-LDL was prepared and identified, and HUVECs were cultured. After 2-hour intervention of different drugs and 24-hour following intervention of Ox-LDL, the number of HUVECs was observed by phase contrast optical microscope and the activity of the HUVECs was observed by methyl thiazolyl tetrazolium （MTT） technique. Superoxide dismutase （SOD） activity and nitric oxide （NO） content were assayed by respective kit. The protein expressions and mRNA levels of peroxisome proliferators activated receptor γ （PPAR-γ） and endothelial nitric oxide synthase （eNOS） were measured by western blot technique and reverse transcription polymerase chain reaction （RT-PCR）. Results： Ox-LDL could increase the apoptosis rate of the HUVECs and decrease the NO release as compared with the blank control group （P〈0.05）. These effects induced by Ox-LDL were all significantly inhibited by Huaxia shallot preparation. It could up-regulate the protein expressions and mRNA levels of PPAR-γ and eNOS significantly （P〈0.05）. Huaxia shallot preparation could decrease the apoptosis rate of the HUVECs. Conclusion： Ox-LDL may be involved in the initiation and progression of atherosclerosis by injuring the endothelial cells directly and may cause the endothelial dysfunction. Huaxia shallot preparation can protect against Ox-LDL induced endothelial cell injury by up-regulating the protein expressions and mRNA levels of PPAR-γ and eNOS. It suggests that Huaxia shallot preparation may play a role in the prevention and treatment of cardiovascular disease.

关 键 词：动脉粥样硬化 内皮 低密度脂蛋白类 一氧化氯合酶 过氧化物酶类 小葱 

分 类 号：R96[医药卫生—药理学]