P3肽抑制RAW264.7巨噬细胞泡沫化的分子机制  

Peptide P3 ameliorates foam cell formation in RAW264.7 cells and its molecular mechanisms

作　　者：李佳慧 蒋宇恒 王承瑾 江林 邓艳 张礼林 许庆忠 李红梅 LI Jia-hui;JIANG Yu-heng;WANG Cheng-jin;JIANG Lin;DENG Yan;ZHANG Li-lin;XU Qing-zhong;LI Hong-mei(Dept of Biochemistry and Molecular Biology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学基础医学院生物化学与分子生物学教研室,贵州贵阳550025

出　　处：《中国药理学通报》2023年第9期1711-1717,共7页Chinese Pharmacological Bulletin

基　　金：贵州省科学技术基金资助项目(黔科合基础-ZK[2021]一般104);贵州省卫生健康委科学基金资助项目(No gzwkj2023-252);大学生创新创业训练计划项目资助(No 202010660022)。

摘　　要：目的观察P3肽对RAW264.7巨噬细胞脂质沉积的影响,并探讨其作用机制。方法采用MTT法筛选P3肽及氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)的作用浓度,并用80 mg·L^(-1)的ox-LDL诱导RAW264.7细胞形成泡沫细胞;分别采用油红O染色和总胆固醇含量测定试剂盒,检测细胞内脂质沉积及总胆固醇含量;实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)及Western blot检测三磷酸腺苷结合盒转运体A1(ATP-binding cassette transporter A1,ABCA1)、三磷酸腺苷结合盒转运体G1(ATP-binding cassette transporter G1,ABCG1)的mRNA和蛋白表达变化;分别应用肝X受体(liver X receptor,LXR)的激动剂T0901317、GW3965,以及LXR抑制剂GSK2033进一步验证P3肽调控ABCA1、ABCG1表达的作用机制。结果P3肽明显减少RAW264.7细胞内脂质沉积,降低细胞内总胆固醇含量,上调ABCA1、ABCG1mRNA和蛋白表达;P3肽上调ABCA1、ABCG1的作用与LXR激动剂T0901317、GW3965相似;加入抑制LXR基因转录的抑制剂GSK2033后,P3肽对ABCA1、ABCG1的基因表达无上调作用。结论P3肽可减少RAW264.7巨噬细胞内脂质积聚,抑制泡沫细胞的形成,其作用机制可能与激活LXR-ABCA1/ABCG1通路有关。Aim To examine the effect of peptide P3 on lipid accumulation in RAW264.7 cells and the underlying molecular mechanism.Methods MTT method was used to screen the concentration of peptide P3 and oxidized low density lipoprotein(ox-LDL),and RAW.264.7 cells were induced to form foam cells by ox-LDL with 80 mg·L^(-1).Intracellular lipid accumulation and total cholesterol content were detected by oil red O staining and colorimetry respectively.The mRNA and protein levels of ATP-binding cassette transporter A1(ABCA1)and ATP-binding cassette transporter G1(ABCG1)were measured by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis.Liver X receptor(LXR)agonist T0901317,GW3965 and LXR inhibitor GSK2033 were used to further verify the mechanism of peptide P3 regulation the expression of ABCA1 and ABCG1.Results Peptide P3 significantly reduced lipid deposition and decreased total cholesterol content in RAW264.7 cells.Peptide P3 significantly up-regulated the relative levels of mRNA and protein expression of ABCA1 and ABCG1.Compared with LXR agonists,peptide P3 up-regulated ABCA1 and ABCG1 in a similar way.After addition of GSK2033,an inhibitor of LXR gene transcription,P3 peptide had no up-regulatory effect on ABA1 and ABCG1 gene expression.Conclusion Peptide P3 can reduce lipid accumulation and suppress the formation of foam cells in RAW264.7 cells,which might be related to activating the LXR-ABCA1/ABCG1 pathway.

关 键 词：P3肽 RAW264.7细胞 脂质沉积 泡沫细胞 三磷酸腺苷结合盒转运体A1 三磷酸腺苷结合盒转运体G1 肝X受体 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学] R543.5R589.2R977.6