Zhang Zhongbai, Qin Xueting, Liu Jiyuan, Li Yanchun, Chen Huaxin, Xie Hongwei, Chen Jingxun, Li Chuang, Tong Yang, Yang Min, Zhang Mei
The Fourth Detachment, China Coast Guard, Wenchang, Hainan 571300, P.R. China.
Department of Nephrology with Integrated Traditional Chinese and Western Medicine, No. 2 People's Hospital of The Three Gorges University, Yichang, Hubei 443000, P.R. China.
Exp Ther Med. 2022 Jul 19;24(3):579. doi: 10.3892/etm.2022.11516. eCollection 2022 Sep.
Coronary atherosclerotic heart disease poses a significant threat to human health. The pathological basis is atherosclerosis and foam-cell formation is the key factor in the initiation of atherosclerosis. In the present study, foam cell models were established using 50 ng/ml oxidized low-density lipoprotein to stimulate cultures of THP-1 cells for 72 h. The expression of zinc finger protein 580 (ZNF580), a Cys2-His2 zinc finger protein containing 172 amino acids that was originally cloned by screening a human aortic cDNA library, was measured in foam cells and its interaction with various regulatory factors during foam-cell formation was investigated. Oil red O staining was used to observe cell morphology and intracellular lipid levels. Lentivirus transfection was employed to either overexpress or silence ZNF580 in THP-1 cells, and an inverted fluorescence microscope was used to observe the distribution of ZNF580 immunofluorescence to determine the transfection rate. RNA and total protein were extracted and the expression levels of ZNF580, CD36, peroxisome proliferator-activated receptor-γ (PPAR-γ), ATP-binding cassette transporter A1 (ABCA1) and apolipoprotein E (ApoE) were measured by reverse transcription-quantitative PCR. The protein levels were examined by western blot analysis to evaluate the interaction between ZNF580 and associated regulatory factors. ZNF580 was able to significantly increase the expression levels of ApoE and ABCA1 and significantly decrease the expression levels of CD36 and PPAR-γ, suggesting that ZNF580-mediated inhibition of foam-cell formation is associated with the PPAR-γ-CD36 signalling pathway. Based on these findings, ZNF580 may be a potential therapeutic candidate for the treatment of coronary atherosclerotic heart disease.
冠状动脉粥样硬化性心脏病对人类健康构成重大威胁。其病理基础是动脉粥样硬化，而泡沫细胞形成是动脉粥样硬化起始的关键因素。在本研究中，使用50 ng/ml氧化型低密度脂蛋白刺激THP-1细胞培养物72小时，建立泡沫细胞模型。检测了锌指蛋白580（ZNF580）的表达，ZNF580是一种含有172个氨基酸的Cys2-His2锌指蛋白，最初通过筛选人主动脉cDNA文库克隆得到，并研究了其在泡沫细胞形成过程中与各种调节因子的相互作用。采用油红O染色观察细胞形态和细胞内脂质水平。利用慢病毒转染在THP-1细胞中过表达或沉默ZNF580，并使用倒置荧光显微镜观察ZNF580免疫荧光的分布以确定转染率。提取RNA和总蛋白，通过逆转录定量PCR检测ZNF580、CD36、过氧化物酶体增殖物激活受体γ（PPAR-γ）、ATP结合盒转运蛋白A1（ABCA1）和载脂蛋白E（ApoE）的表达水平。通过蛋白质印迹分析检测蛋白质水平，以评估ZNF580与相关调节因子之间的相互作用。ZNF580能够显著增加ApoE和ABCA1的表达水平，并显著降低CD36和PPAR-γ的表达水平，提示ZNF580介导的对泡沫细胞形成的抑制作用与PPAR-γ-CD36信号通路有关。基于这些发现，ZNF580可能是治疗冠状动脉粥样硬化性心脏病的潜在候选药物。
