microRNA-125a-5p对巨噬细胞THP-1泡沫化过程的影响及其机制
发布时间：2018-12-12 20:02
【摘要】：目的探讨microRNA-125a-5p(miR-125a-5p)对巨噬细胞THP-1泡沫化过程的影响及其机制。方法在Lipao2000的介导下,将miR-125a-5p的增强剂(mimics)/抑制剂(inhibition)(终浓度均为50 nmol/L)分别转染THP-1细胞24 h后,加入终浓度为100 mg/L的低密度脂蛋白(oxygenized-low density lipoprotein,ox-LDL),继续培养48 h,同时设空白对照组(未转染)及ox-LDL组(仅加入终浓度为100 mg/L的ox-LDL)。采用油红O染色和脂质染色的半定量分析法检测miR-125a-5p对THP-1细胞泡沫化的影响;Cholesterol/Cholesteryl Ester Quantitation Kit检测miR-125a-5p对THP-1细胞泡沫化过程中脂质蓄积的影响;Real-time PCR和Western blot法检测miR-125a-5p对THP-1细胞中酰基辅酶A胆固醇酰基转移酶-1(acyl-coenzyme A:cholesterol acyltrasferase-1,ACAT-1)、ATP结合盒转运体A1(ATP binding casstte transporter A1,ABCA1)及细胞清道夫受体-A1(scavenger receptor-A1,SR-A1)mRNA及蛋白表达水平的影响。结果与ox-LDL组比较,ox-LDL+mimics组巨噬细胞泡沫化程度及脂质蓄积量均明显下降(P均0.05),ox-LDL+inhibition组均明显增加(P均0.05);ox-LDL+mimics组ACAT-1和SR-A1 mRNA及蛋白表达水平明显降低(P均0.05),而ABCA1 mRNA及蛋白表达水平明显升高(P均0.05);ox-LDL+inhibition组ACAT-1和SR-A1 mRNA及蛋白表达水平明显增加(P均0.05),而ABCA1 mRNA及蛋白表达水平明显降低(P均0.05)。结论 miR-125a-5p可通过调解ACAT-1、ABCA1和SR-A1的表达水平,减少巨噬细胞内脂质类的堆积,抑制巨噬细胞向泡沫细胞的转化,从而发挥抗动脉粥样硬化(atheroscler-osis,As)的作用。
[Abstract]:Objective to investigate the effect and mechanism of microRNA-125a-5p (miR-125a-5p) on THP-1 foaming of macrophages. Methods THP-1 cells were transfected with miR-125a-5p enhancer (mimics) / (inhibition) (50 nmol/L) mediated by Lipao2000 for 24 h. Oxygenized-low density lipoprotein, with final concentration of 100 mg/L was added. Ox-LDL), cultured for 48 h, with blank control group (untransfected) and ox-LDL group (ox-LDL with final concentration of 100 mg/L). Oil red O staining and lipid staining were used to detect the effect of miR-125a-5p on the foaming of THP-1 cells, and Cholesterol/Cholesteryl Ester Quantitation Kit to detect the effect of miR-125a-5p on the accumulation of lipid in THP-1 cells. Real-time PCR and Western blot methods were used to detect the effect of miR-125a-5p on the expression of acyl-coenzyme A:cholesterol acyltrasferase-1,ACAT-1), ATP binding cassette transporter A1 (ATP binding casstte transporter A1 in THP-1 cells. ABCA1) and the expression of scavenger receptor-A1 (scavenger receptor-A1,SR-A1) mRNA and protein. Results compared with ox-LDL group, the degree of macrophage foaming and the amount of lipid accumulation in ox-LDL mimics group were significantly decreased (P 0.05), while those in ox-LDL inhibition group were significantly increased (P 0.05). In ox-LDL mimics group, the expression of ACAT-1 and SR-A1 mRNA and protein decreased significantly (P 0.05), while the expression of ABCA1 mRNA and protein increased significantly (P 0.05). In ox-LDL inhibition group, the expression of ACAT-1 and SR-A1 mRNA and protein increased significantly (P 0.05), while the expression of ABCA1 mRNA and protein decreased significantly (P 0.05). Conclusion miR-125a-5p can mediate the expression of ACAT-1,ABCA1 and SR-A1, reduce the accumulation of lipids in macrophages, inhibit the transformation of macrophages to foam cells, and thus play a role in anti-atherosclerosis (atheroscler-osis,). As).
【作者单位】： 辽宁医学院附属第一医院心外科;
【基金】：国家自然科学基金项目(81171111)
【分类号】：R363
[Abstract]:Objective to investigate the effect and mechanism of microRNA-125a-5p (miR-125a-5p) on THP-1 foaming of macrophages. Methods THP-1 cells were transfected with miR-125a-5p enhancer (mimics) / (inhibition) (50 nmol/L) mediated by Lipao2000 for 24 h. Oxygenized-low density lipoprotein, with final concentration of 100 mg/L was added. Ox-LDL), cultured for 48 h, with blank control group (untransfected) and ox-LDL group (ox-LDL with final concentration of 100 mg/L). Oil red O staining and lipid staining were used to detect the effect of miR-125a-5p on the foaming of THP-1 cells, and Cholesterol/Cholesteryl Ester Quantitation Kit to detect the effect of miR-125a-5p on the accumulation of lipid in THP-1 cells. Real-time PCR and Western blot methods were used to detect the effect of miR-125a-5p on the expression of acyl-coenzyme A:cholesterol acyltrasferase-1,ACAT-1), ATP binding cassette transporter A1 (ATP binding casstte transporter A1 in THP-1 cells. ABCA1) and the expression of scavenger receptor-A1 (scavenger receptor-A1,SR-A1) mRNA and protein. Results compared with ox-LDL group, the degree of macrophage foaming and the amount of lipid accumulation in ox-LDL mimics group were significantly decreased (P 0.05), while those in ox-LDL inhibition group were significantly increased (P 0.05). In ox-LDL mimics group, the expression of ACAT-1 and SR-A1 mRNA and protein decreased significantly (P 0.05), while the expression of ABCA1 mRNA and protein increased significantly (P 0.05). In ox-LDL inhibition group, the expression of ACAT-1 and SR-A1 mRNA and protein increased significantly (P 0.05), while the expression of ABCA1 mRNA and protein decreased significantly (P 0.05). Conclusion miR-125a-5p can mediate the expression of ACAT-1,ABCA1 and SR-A1, reduce the accumulation of lipids in macrophages, inhibit the transformation of macrophages to foam cells, and thus play a role in anti-atherosclerosis (atheroscler-osis,). As).
【作者单位】： 辽宁医学院附属第一医院心外科;
【基金】：国家自然科学基金项目(81171111)
【分类号】：R363
【参考文献】
相关期刊论文 前1条
1 陈妍梅;李浪;;microRNA与动脉粥样硬化炎症反应[J];中国动脉硬化杂志;2010年08期
【共引文献】
相关期刊论文 前5条
1 陆永光;李浪;陈妍梅;钟继明;曾书q，
本文编号：2375171
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2375171.html
最近更新
教材专著
