Department of Cardiology, Henan Provincial People's Hospital, Zhengzhou, Henan, People's Republic of China.
J Cell Biochem. 2018 Feb;119(2):1911-1921. doi: 10.1002/jcb.26352. Epub 2017 Nov 10.
This study is designed to determine whether lincRNA-DYNLRB2-2 could promote cholesterol efflux through regulating the expression of TLR2. THP-1 and RAW264.7 cells were incubated with oxLDL for 48 h to induce the formation of foam cells, and ORO staining was performed and intracellular cholesterol contents were measured by HPLC assay. qRT-PCR and Western blotting were performed to detect mRNA and protein expression levels, respectively. Lentiviral vector LV-DYNLRB2-2 and lincRNA-DYNLRB2-2 siRNA was constructed to explore its potential role. The cholesterol efflux was assessed by liquid scintillation counting. The effects of TRL2 were determined in apoE mice that fed a high fat diet and were randomly divided into three groups and infected with LV-Mock, LV-Sh-TRL2, or LV-TRL2. Atherosclerosis was observed in the aortic sinus and the levels of cytokines and serum biochemical parameters were measured. Ox-LDL induced foam cell formation in the THP-1 and RAW264.7 cells. LincRNA DYN-LRB2-2 was upregulated in oxLDL-treated THP-1 and Raw264.7 cells. LincRNA-DYNLRB2-2 plays important role in regulating the cholesterol efflux, ABCA1 expression level and anti-inflammatory processes in THP-1 and RAW264.7 cells. Further study indicated that lincRNA-DYNLRB2-2 negatively regulated TRL2 expression and TRL2 overexpression reversed the effects of lincRNA-DYNLRB2-2 on cholesterol efflux and ABCA1 expression level in THP-1 and RAW264.7 cells. Besides, we found TRL2 plays important role in lipid accumulation, plaque formation and regulating serum inflammatory cytokines level in apoE mice with a high fat diet. LincRNA DYN-LRB2-2 upregulates cholesterol efflux by decreasing TLR2 expression in macrophages.
本研究旨在探讨 lincRNA-DYNLRB2-2 是否可通过调节 TLR2 的表达促进胆固醇外流。将 THP-1 和 RAW264.7 细胞用 oxLDL 孵育 48 小时以诱导泡沫细胞形成，并用 ORO 染色，并通过 HPLC 测定法测量细胞内胆固醇含量。进行 qRT-PCR 和 Western blot 分别检测 mRNA 和蛋白表达水平。构建慢病毒载体 LV-DYNLRB2-2 和 lincRNA-DYNLRB2-2 siRNA 以探索其潜在作用。通过液体闪烁计数评估胆固醇外流。在高脂饮食喂养的 apoE 小鼠中确定 TRL2 的作用，并将其随机分为三组，感染 LV-Mock、LV-Sh-TRL2 或 LV-TRL2。观察主动脉窦中的动脉粥样硬化，并测量细胞因子和血清生化参数的水平。oxLDL 诱导 THP-1 和 RAW264.7 细胞形成泡沫细胞。LincRNA DYN-LRB2-2 在 oxLDL 处理的 THP-1 和 Raw264.7 细胞中上调。LincRNA-DYNLRB2-2 在调节 THP-1 和 RAW264.7 细胞中的胆固醇外流、ABCA1 表达水平和抗炎过程中起重要作用。进一步的研究表明，lincRNA-DYNLRB2-2 负调控 TLR2 的表达，TLR2 过表达逆转了 lincRNA-DYNLRB2-2 对 THP-1 和 RAW264.7 细胞中胆固醇外流和 ABCA1 表达水平的影响。此外，我们发现 TRL2 在高脂饮食的 apoE 小鼠中脂质积累、斑块形成和调节血清炎症细胞因子水平方面发挥重要作用。LincRNA DYN-LRB2-2 通过降低巨噬细胞中 TLR2 的表达来增加胆固醇外流。
