ILF3 is responsible for hyperlipidemia-induced arteriosclerotic calcification by mediating BMP2 and STAT1 transcription
Graphical abstract
Our findings illustrate the working schematic that hyperlipidemia-induced ILF3 activation mediates acceleration of atherosclerotic calcification. Hyperlipidemia-increased ILF3 expression mediates BMP2 and STAT1 transcription by directly binding to their promoter regions. ILF3 upregulates BMP2 level and activates Smads signaling to elevate Runx2 transcription. Meanwhile, ILF3 suppresses STAT1 transcription, which promotes Runx2 nuclear translocation and regulates osteogenic differentiation. In addition, ILF3-mediated hyperlipidemia induces a phenotypic switch of VSMCs from contractile to a dedifferentiated synthetic phenotype and macrophages to a pro-inflammatory M1 phenotype, which in turn aggravates VSMC calcification to promote atherosclerotic calcification.
Introduction
Vascular calcification is a common phenomenon in many physiological and pathological diseases including aging, end-stage renal disease, diabetes mellitus and cardiovascular diseases [1,2]. Vascular calcification can occur in different locations of the vessel wall including intima and media but exists mainly in intimal layers in atherosclerosis and can induce atherosclerotic plaque susceptibility and further lead to myocardial infarction, plaque rupture and stroke [3].
Recent studies suggested that vascular calcification is an active cell regulatory process characterized by the involvement of various cells such as vascular smooth muscle cells (VSMCs), pericytes, myofibroblasts, macrophages, vascular mesenchymal progenitors and endothelial cells [[4], [5], [6], [7]]. Under multiple pro-calcific stimuli, VSMCs can undergo a phenotype switch from a contractile to osteoblastic phenotype accompanied by loss of contractile markers (smooth muscle 22 alpha [SM22Ī±], calponin and alpha smooth muscle actin [Ī±-SMA]) and an increase in levels of bone-related genes (runt-related transcription factor 2 [Runx2], osteopontin [OPN], osteocalcin, bone morphogenetic protein 2 [BMP2], and Msx and become the main source of osteoblastic cells, which leads to vascular calcification. In addition, macrophages can undergo a phenotype shift and participate in atherosclerotic calcification [5,8]. Because of the diversity and complexity of calcification mechanisms, ideal drugs preventing or reversing atherosclerotic calcification are unavailable. The underlying molecular mechanisms of atherosclerotic calcification still need further study.
Interleukin enhancer-binding factor 3 (ILF3), as a double-stranded RNA (dsRNA)-binding protein, combines with other proteins, mRNAs, small noncoding RNAs, and dsRNAs to regulate transcription, translation, mRNA stability and noncoding RNA biogenesis [9]. In the cardiovascular system, ILF3 can inhibit myocardial hypertrophy [10]. Also, the association between ILF3 and myocardial infarction is affected by low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol metabolism, which indicates interactions between genes [11]. Recent studies have reported insights into the possible physiological roles of ILF3 in stroke, inflammation, and dyslipidaemia, but its role in vascular calcification has not been reported.
In this study, we used human samples and murine models with conditional ILF3 knockout and overexpression in VSMCs and macrophages to explore the roles of ILF3 in atherosclerotic calcification.
Section snippets
Body weight (BW) and serum indexes in mice
Mice BW and serum levels of TC, TG, BG, HDL-C, LDL-C, calcium and phosphorus are in Tables 1 and 2. Both ApoEā/-ILF3SM-KO and ApoEā/-ILF3M-KO mice showed lower BW and TC, TG and LDL-C levels but higher HDL-C level than ApoEā/ā mice (*PĀ <Ā 0.05). For all ILF3-overexpressed mice, BW and TC, TG and LDL-C levels were higher but HDL-C level was lower relative to ApoEā/āmice (*PĀ <Ā 0.05). BG, calcium and phosphorus levels did not differ between groups. The one that plays the most main role in
Discussion
ILF3 has been verified to play important roles in dyslipidemia and the cardiovascular system [10,11]. However, whether ILF3 is linked to dyslipidemia-induced atherosclerotic calcification has not been reported. In the present study, we used ILF3 conditional genetic deletion and transgenic mouse models to investigate the role of ILF3 in atherosclerotic calcification. Hyperlipidemia could augment ILF3 expression in calcified VSMCs and macrophages and in atherosclerotic calcification in humans and
Human coronary artery samples
Atherosclerotic and control epicardial coronary artery segments were from human specimens with atherosclerotic disease. The specimens were donated by the Shandong Red Cross Society.The experiment protocols were examined and approved by the review committee of Qilu Hospital, Jinan, China(ethics approval No. KYLL-2018(KS)-233).
Animals
The study was conducted in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The
Declaration of Competing Interest
The authors declare that they have no conflict of interest.
Acknowledgments
This work was supported by the National Natural Science Foundation of China (#91839301 and #81970251), the National key R & D program of China (#2017YFC0908700, 2017YFC0908703) and the Taishan Scholar Project of Shandong Province of China (No. ts20190972).
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