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. 2022 Jul 28;20(1):337.
doi: 10.1186/s12967-022-03542-0.

Asprosin inhibits macrophage lipid accumulation and reduces atherosclerotic burden by up-regulating ABCA1 and ABCG1 expression via the p38/Elk-1 pathway

Jin Zou  1 , Can Xu  1 , Zhen-Wang Zhao  2 , Shan-Hui Yin  3 , Gang Wang  4
Affiliations

Asprosin inhibits macrophage lipid accumulation and reduces atherosclerotic burden by up-regulating ABCA1 and ABCG1 expression via the p38/Elk-1 pathway

Jin Zou et al. J Transl Med. .

Abstract

Background: Asprosin, a newly discovered adipokine, is a C-terminal cleavage product of profibrillin. Asprosin has been reported to participate in lipid metabolism and cardiovascular disease, but its role in atherogenesis remains elusive.

Methods: Asprosin was overexpressed in THP-1 macrophage-derived foam cells and apoE-/- mice using the lentiviral vector. The expression of relevant molecules was determined by qRT-PCR and/or western blot. The intracellular lipid accumulation was evaluated by high-performance liquid chromatography and Oil red O staining. HE and Oil red O staining was employed to assess plaque burden in vivo. Reverse cholesterol transport (RCT) efficiency was measured using [3H]-labeled cholesterol.

Results: Exposure of THP-1 macrophages to oxidized low-density lipoprotein down-regulated asprosin expression. Lentivirus-mediated overexpression of asprosin promoted cholesterol efflux and inhibited lipid accumulation in THP-1 macrophage-derived foam cells. Mechanistic analysis revealed that asprosin overexpression activated p38 and stimulated the phosphorylation of ETS-like transcription factor (Elk-1) at Ser383, leading to Elk-1 nuclear translocation and the transcriptional activation of ATP binding cassette transporters A1 (ABCA1) and ABCG1. Injection of lentiviral vector expressing asprosin diminished atherosclerotic lesion area, increased plaque stability, improved plasma lipid profiles and facilitated RCT in apoE-/- mice. Asprosin overexpression also increased the phosphorylation of p38 and Elk-1 as well as up-regulated the expression of ABCA1 and ABCG1 in the aortas.

Conclusion: Asprosin inhibits lipid accumulation in macrophages and decreases atherosclerotic burden in apoE-/- mice by up-regulating ABCA1 and ABCG1 expression via activation of the p38/Elk-1 signaling pathway.

Keywords: ABCA1; ABCG1; Asprosin; Atherosclerosis; Elk-1; p38.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Asprosin prevents macrophages from lipid accumulation. A After incubation of THP-1 macrophages with or without 50 µg/mL ox-LDL for 48 h, qRT-PCR and western blot were used to detect asprosin expression (n = 3). B–D THP-1 macrophage-derived foam cells were transfected with LV-Mock or LV-Asprosin for 72 h (n = 3). B Protein samples were immunoblotted with antibodies against asprosin or β-actin. C The contents of intracellular TC, FC and CE were measured by HPLC. D Representative images of Oil red O staining. Data are expressed as mean ± SD. **P < 0.01, ***P < 0.001
Fig. 2
Fig. 2
Asprosin promotes ABCA1 and ABCG1 expression and cholesterol efflux from macrophages. A–E THP-1 macrophage-derived foam cells were transduced with LV-Mock or LV-Asprosin for 72 h (n = 3). A The expression of ABCA1 and ABCG1 was determined by qRT-PCR and western blot. B, C Representative fluorescent images of NBD-cholesterol burden and quantitative analyses of cholesterol efflux to apoA-I and HDL. D The qRT-PCR and western blot assays of CD36 and SR-A expression. E Representative fluorescent images and quantification of Dil-ox-LDL uptake. Data presented are mean ± SD. ***P < 0.001. NS indicates not significant
Fig. 3
Fig. 3
Elk-1 phosphorylation at Ser383 is involved in asprosin-induced up-regulation of ABCA1 and ABCG1 expression. A–E THP-1 macrophage-derived foam cells were treated with LV-Mock or LV-Asprosin for 72 h (n = 3). A Detection of LXRα expression using qRT-PCR and western blot. B The expression of PCSK9 was measured by using qRT-PCR and western blot. C The mRNA and protein levels of Elk-1 were determined by qRT-PCR and western blot, respectively. D Western blot analysis of Elk-1 protein expression in the cytoplasm and nucleus. E Measurement of Elk-1 phosphorylation at Ser383, Ser389 and Thr417 by western blot. F–H THP-1 macrophage-derived foam cells were transfected with Elk-1S383A through lentiviral vector for 72 h, followed by treatment with or without LV-Asprosin for another 72 h (n = 3). F The qRT-PCR and western blot analyses of ABCA1 and ABCG1 expression. G, H Representative fluorescent images of NBD-cholesterol burden along with quantitative analyses of cholesterol efflux mediated by apoA-I and HDL. Data shown are mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. NS indicates not significant
Fig. 4
Fig. 4
Asprosin stimulates Elk-1 phosphorylation and ABCA1 and ABCG1 expression by activating p38. A After 72 h of transfection with LV-Mock or LV-Asprosin, cell lysates were immunoblotted with indicated antibodies (n = 3). B–F THP-1 macrophage-derived foam cells were treated with SB203580 for 6 h and then transfected with or without LV-Asprosin for an additional 72 h (n = 3). B Detection of Elk-1 phosphorylation at Ser383 using western blot. C Western blot analysis of nuclear and cytoplasmic Elk-1 protein expression. D Measurement of ABCA1 and ABCG1 expression by qRT-PCR and western blot. E, F Representative fluorescent images of NBD-cholesterol burden and quantification of cholesterol efflux to apoA-I and HDL. Data represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. NS indicates not significant
Fig. 5
Fig. 5
Asprosin inhibits the development of atherosclerosis. A–G ApoE−/− mice were fed a Western diet for 12 weeks and simultaneously underwent tail vein injection of LV-Mock or LV-Asprosin once three weeks (n = 15 mice per group). A Comparison of body weight gain (n = 15 mice per group). B Western blot analysis of asprosin protein expression in the aortas (n = 5 mice per group). C Representative images of aortic arch regions with white plaques (green arrows). D Enface analysis of atherosclerotic lesion area using Oil Red O staining (n = 5 mice per group). E–G The aortic valve cross-sections were stained with HE, Oil Red O and Masson Trichrome to evaluate lesion area, lipid deposition and collagen content, respectively (n = 10 mice per group). The results are shown as the mean ± SD. **P < 0.01, ***P < 0.001. NS indicates not significant
Fig. 6
Fig. 6
Asprosin ameliorates plasma lipid profile and enhances RCT. A–D Circulating levels of TC, LDL-C, HDL-C and TG were measured by the enzymatic methods (n = 10 mice per group). E J774 macrophages loaded with ac-LDL and [3H]-cholesterol were injected into apo E−/− mice. The radioactivity of [3H]-cholesterol in the plasma, liver and feces was determined by a liquid scintillation counter (n = 5 mice per group). F ELISA was used to detect plasma PCSK9 levels (n = 10 mice per group). Data are given as the mean ± SD. *P < 0.05, ***P < 0.001. NS indicates not significant
Fig. 7
Fig. 7
Asprosin promotes p38 and Elk-1 phosphorylation and ABCA1 and ABCG1 expression in the aortas. A Protein samples were immunoblotted with antibodies against p-p38, p38 or β-actin in the aortas (n = 5 mice per group). B Measurement of aortic Elk-1 expression by qRT-PCR and western blot (n = 5 mice per group). C Detection of aortic Elk-1 phosphorylation at Ser383 using western blot (n = 5 mice per group). D Western blot analysis of aortic Elk-1 protein levels in the cytoplasm and nucleus (n = 5 mice per group). E The qRT-PCR and western blot assays of ABCA1 and ABCG1 expression in the aortas (n = 5 mice per group). The results are shown as the mean ± SD. **P < 0.01, ***P < 0.001. NS indicates not significant
Fig. 8
Fig. 8
A model describing the mechanism for asprosin-induced atheroprotection. Asprosin activates p38 and then phosphorylates Elk-1 at Ser383. After phosphorylation, Elk-1 enters the nucleus to stimulate ABCA1 and ABCG1 transcription. Increased ABCA1 and ABCG1 expression promotes cholesterol efflux from macrophages, leading to acceleration of RCT and alleviation of atherosclerotic plaque burden
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