硒甲基硒代半胱氨酸对THP-1细胞源性泡沫细胞炎症因子表达的影响及其机制  

Effect of L-Se-methylselenocysteine on expression of inflammatory factors in THP-1 cell-derived foam cells and its mechanism

作　　者：陈丹 袁林 高星星 夏燕 张宗星 李玮怡 汪莲开 CHEN Dan;YUAN Lin;GAO Xingxing;XIA Yan;ZHANG Zongxing;LI Weiyi;WANG Liankai(Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China;Hubei Provincial Clinical Medical Research Center for Nephropathy,Enshi 445000,China;Health Science Center,Hubei Minzu University,Enshi 445000,China)

机构地区：[1]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [2]湖北省肾脏病临床医学研究中心,湖北恩施445000 [3]湖北民族大学医学部,湖北恩施445000

出　　处：《中国病理生理杂志》2024年第12期2218-2225,共8页Chinese Journal of Pathophysiology

基　　金：恩施州科技计划项目(No.XYJ2021000042);湖北民族大学附属民大医院开放基金项目(No.OIR202304Q);风湿性疾病发生与干预湖北省重点实验室开放基金项目(No.PT022313)。

摘　　要：目的:探究硒甲基硒代半胱氨酸(L-SeMSC)抑制人单核白血病细胞THP-1源性巨噬细胞泡沫化的作用机制。方法:先用佛波酯(PMA)诱导THP-1细胞源性巨噬细胞,再用氧化低密度脂蛋白(ox-LDL)诱导THP-1巨噬细胞源性泡沫细胞模型。CCK-8法和流式细胞术检测不同浓度L-SeMSC对THP-1细胞活性和凋亡的影响;油红O染色观察脂滴的沉积;并通过qRT-PCR法检测白细胞介素1β(IL-1β)、细胞间黏附分子1(ICAM-1)、脂肪酸转位酶(FAT/CD36)、溶质载体家族27成员4(SLC27A4)和超氧化物歧化酶1(SOD1)的mRNA水平;Western blot检测IL-6、IL-1β、肿瘤坏死因子α(TNF-α)、基质金属蛋白酶9(MMP9)及核因子κB(NF-κB)信号相关蛋白水平。结果:25~200μmol/L的L-SeMSC增加细胞活力,不同浓度的L-SeMSC对细胞凋亡没有显著影响。与ox-LDL组相比,LSeMSC组细胞脂滴沉积明显减少;CD36、SLC27A4、IL-1β和ICAM-1的mRNA水平显著降低,同时抗氧化酶SOD1的mRNA水平明显增加;IL-1β、IL-6、TNF-α、MMP9及磷酸化p65(p-p65)蛋白水平均明显减少。结论:L-SeMSC可能具有抗动脉粥样硬化作用,其通过降低炎性因子的水平、抑制泡沫化的发生、减少巨噬细胞脂滴的形成,相关抗炎机制可能主要是通过调控NF-κB信号通路中的关键蛋白表达。AIM:To investigate the mechanism by which L-Se-methylselenocysteine(L-SeMSC)inhibits foam cell formation in THP-1-derived macrophages from human monocytic leukemia cells.METHODS:THP-1-derived macrophages were first induced using phorbol 12-myristate 13-acetate(PMA),followed by the use of oxidized low-density lipoprotein(ox-LDL)to establish a foam cell model.The effects of various concentrations of L-SeMSC on THP-1 cell viability and apoptosis were assessed using the CCK-8 assay and flow cytometry.Lipid droplet accumulation was visualized through oil red O staining.Additionally,the mRNA levels of interleukin-1β(IL-1β),intercellular adhesion molecule 1(ICAM-1),fatty acid translocase(FAT/CD36),solute carrier family 27 member 4(SLC27A4),and superoxide dismutase 1(SOD1)were measured using qRT-PCR.The levels of inflammatory proteins,including IL-6,IL-1β,tumor necrosis factor-α(TNF-α),matrix metalloproteinase 9(MMP9),and nuclear factor-κB(NF-κB)signaling-related proteins in activated B cells were assessed via Western blot.RESULTS:Treatment with L-SeMSC at concentrations ranging from 25 to 200μmol/L enhanced cell viability,with no significant impact on apoptosis observed across the different concentra‑tions. The accumulation of lipid droplets was notably reduced in the L-SeMSC group compared to the ox-LDL group. The mRNA levels of lipid transport-related genes (CD36 and SLC27A4) and inflammation-related genes (IL-1β and ICAM-1) were significantly decreased, while SOD1 mRNA levels increased significantly. Furthermore, the expression levels of in-flammatory proteins IL-1β, IL-6, TNF-α, MMP9, and phosphorylated p65 (p-p65) were markedly reduced. CONCLU⁃ SION: L-SeMSC may exert anti-atherosclerotic effects by decreasing inflammatory factor levels, inhibiting foam cell for-mation, and reducing lipid droplet accumulation in macrophages. The associated anti-inflammatory mechanism appears to primarily involve the modulation of key proteins in the NF-κB signaling pathway.

关 键 词：硒甲基硒代半胱氨酸 动脉粥样硬化 炎症因子 脂滴形成 

分 类 号：R714.252[医药卫生—妇产科学] R364.5[医药卫生—临床医学] Q28[生物学—细胞生物学]