LncRNA H19在THP-1来源泡沫细胞脂质蓄积中的作用及机制  

Role and mechanism of LncRNA H19 in lipid accumulation of THP-1-derived foam cells

作　　者：阿曼古丽·如则 王雪梅 尼鲁帕尔·谢甫开提 赵翎 赵帮豪 霍强 高晓明[1,2] Amanguli Ruze;WANG Xuemei;Nilupaer Xiepukaiti;ZHAO Ling;ZHAO Banghao;HUO Qiang;GAO Xiaoming(State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia;Xinjiang Key Laboratory of Medical Animal Model Research,Xinjiang Medical University,Urumqi 830054,China;Department of Hygiene,School of Public Health,Xi’an Medical University,Xi’an 710021,China;Department of Cardiology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]省部共建中亚高发病成因与防治国家重点实验室 [2]新疆医学动物模型研究重点实验室,乌鲁木齐830054 [3]西安医学院公共卫生学院卫生学教研室,西安710021 [4]新疆医科大学第一附属医院心脏中心,乌鲁木齐830054

出　　处：《新疆医科大学学报》2023年第1期50-55,共6页Journal of Xinjiang Medical University

基　　金：新疆重点实验室开放课题项目(2018D04030);国家自然科学基金地区基金项目(82060073)。

摘　　要：目的探讨长链非编码RNA(Long non-coding RNA,LncRNA)H19在人单核细胞THP-1源巨噬细胞脂质蓄积中的作用及可能的分子机制。方法THP-1细胞经佛波酯(PMA)和氧化型低密度脂蛋白(ox-LDL)孵育诱导成为泡沫细胞,构建体外动脉粥样硬化(泡沫细胞)模型。利用Lipofectamine TM 3000转染试剂,将H19小干扰RNA(siRNA)转染到THP-1细胞以沉默H19基因;油红O染色及异丙醇提取定量的方法检测细胞内脂质蓄积程度;实时荧光定量PCR检测LncRNA H19 mRNA表达水平;Western blot检测过氧化物酶体增殖激活受体(PPAR-α)蛋白表达水平。结果油红O染色及胆固醇检测结果提示泡沫细胞模型构建成功。RT-PCR检测显示,ox-LDL诱导成功的THP-1源泡沫细胞模型中LncRNA H19 mRNA相对表达量明显增加(P<0.01),而在H19 siRNA沉默后,THP-1源泡沫细胞中脂质含量明显减少(P<0.01)。Western blot检测结果发现,ox-LDL刺激后与对照组相比,PPAR-α蛋白表达水平明显降低,而在H19 siRNA沉默组,PPAR-α蛋白表达水平显著回升(P<0.01)。结论LncRNA H19在THP-1来源泡沫细胞中高表达,从而抑制PPAR-α的表达,进而增加THP-1来源巨噬细胞的脂质吞噬,可能成为治疗动脉粥样硬化的潜在靶点。Objective To investigate the role of long noncoding RNA(LncRNA H19)H19 in lipid accumulation of THP-1 macrophages and its possible molecular mechanism.Methods THP-1 cells were incubated with PMA and ox-LDL and induced into foam cells to construct an in vitro atherosclerosis model.Using Lipofectamine 3000 transfection reagent,H19 small interfering RNA(siRNA)was transfected into THP-1 cells to silence H19 gene;the degree of intracellular lipid accumulation was detected by oil red O staining and isopropanol extraction;the expression level of LncRNA H19 mRNA was detected by quantitative real-time PCR,and the expression level of peroxisome proliferation activated receptor(PPAR-α)protein was detected by Western blot.Results The results of oil red O staining and cholesterol detection indicated that the foam cell model was successfully constructed.RT-PCR analysis showed that the relative expression of LncRNA H19 mRNA in THP-1-derived foam cells induced by ox-LDL were significantly increased(P<0.01),while the lipid content in THP-1-derived foam cells were decreased significantly after silencing with H19 siRNA(P<0.01).Western blot analysis showed that after ox-LDL stimulation,the expression level of PPAR-αwas significantly decreased compared with control group,while in H19 siRNA silencing group,the expression level of PPAR-αwas increased significantly(P<0.01).Conclusion LncRNA H19 is highly expressed in THP-1-derived foam cells,thus inhibiting the expression of PPAR-αand further increasing the lipid accumulation of THP-1-derived macrophages,which may become a potential target for the treatment of atherosclerosis.

关 键 词：LncRNA H19 人单核细胞THP-1 PPAR-Α 动脉粥样硬化 

分 类 号：R541.4[医药卫生—心血管疾病]