血凝集素样氧化型低密度脂蛋白受体-1在氧化型低密度脂蛋白诱导的巨噬细胞氧化应激中的作用  

Effects of Lectin-like oxidized low-density lipoprotein receptor-1 on oxidative stress of macrophages induced by oxidized low-density lipoprotein

作　　者：智丽霞 王康[1] 殷利茜 马茴煌 李姚娜 杨慧宇[1] ZHI Lixia;WANG Kang;YIN Lixi;MA Huihuang;LI Yaona;YANG Huiyu(Department of Cardiovascular Medicine,Second Hospital of Shanxi Medical University,Taiyaun 037000,China)

机构地区：[1]山西医科大学第二医院心血管内科,太原037000

出　　处：《国际心血管病杂志》2022年第5期293-298,共6页International Journal of Cardiovascular Disease

基　　金：国家自然科学基金(81700407)。

摘　　要：目的:探讨血凝集素样氧化型低密度脂蛋白受体-1(LOX-1)在氧化型低密度脂蛋白(ox-LDL)诱导的巨噬细胞氧化应激中的作用。方法:将生长至对数增殖期的巨噬细胞随机分成对照组、空质粒对照组(pCon组)、ox-LDL组、LOX-1-小干扰RNA(siRNA)+ox-LDL组,分别测定各组丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性,荧光显微镜及流式细胞仪检测各组巨噬细胞活性氧(ROS)水平,实时定量聚合酶链反应(PCR)及Western blot法检测各组还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX)核心酶NOX2及亚组分p22phox的mRNA及蛋白表达水平。结果:(1)与对照组、pCon组比较,oxLDL组MDA含量明显升高,SOD活性明显下降;与ox-LDL组比较,LOX-1-siRNA+oxLDL组MDA含量明显下降,SOD活性明显升高。(2)荧光显微镜、流式细胞仪检测示oxLDL组巨噬细胞ROS平均荧光强度较正常对照组、pCon组显著增加,而LOX-1-siRNA+ox-LDL组较ox-LDL组显著降低,pCon组与对照组相比差异无统计学意义。(3)实时定量PCR及Western blot检测示ox-LDL组NOX2、p22phox的mRNA和蛋白表达水平较对照组、pCon组显著升高,LOX-1-siRNA+ox-LDL组NOX2、p22phox的m RNA和蛋白表达水平较ox-LDL组则明显下降。结论:LOX-1与ox-LDL作用可使巨噬细胞NOX2、p22phox表达增加,进而促进ROS生成,LOX-1在ox-LDL诱导的巨噬细胞氧化应激中起促进作用。Objective:To evaluate the role of Lectin-like oxidized low-density lipoprotein receptor-1 in oxidative stress of macrophages induced by oxidized low-density lipoprotein(ox-LDL).Methods:Macrophages growing to logarithmic proliferation stage were randomly divided into four groups:control group,empty plasmid control group(pCon group),oxLDL group,and LOX-1-siRNA+ ox-LDL group.Level of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) were determined by biochemical methods,and level of ROS was assessed by fluorescence microscope and flow cytometry.Real-time PCR and western blot were used to detect mRNA and protein expression of NOX2 and p22phox.Results:(1) Compared with control group,levels of MDA were increased and activities of SOD were decreased significantly in ox-LDL group.These changes in MDA levels and SOD activities were more obvious in LOX-1-siRNA+ ox-LDL group.(2) The average fluorescence intensity of ROS was significantly elevated in ox-LDL group compared with control group,while that in LOX-1-siRNA+ ox-LDL group was significantly decreased compared with ox-LDL group.There was no statistical difference between pCon group and control group.(3) The mRNA and protein expression levels of NOX2 and p22phox were significantly higher in ox-LDL group than those in control group and pCon group.These values were significantly reduced in LOX-1-siRNA+ ox-LDL group compared with ox-LDL group.Conclusion:An interaction between LOX-1 and ox-LDL could increase the expression levels of NOX2 and p22phox in macrophages and enhance the production of ROS,suggesting that LOX-1 may play a driving role in ox-LDL-induced oxidative stress in macrophages.

关 键 词：血凝集素样氧化型低密度脂蛋白受体-1 氧化型低密度脂蛋白 巨噬细胞 氧化应激 

分 类 号：R392[医药卫生—免疫学]