| Atherosclerosis is a chronic inflammatory disease owing to lipid metabolism disorders and maladaptive inflammatory response in the large arteries.Atherosclerosis exhibits the characteristics of classical inflammation,such as degeneration,exudation and proliferation.Many inflammatory cells,inflammatory cytokines and mediators,adhesion molecules,chemotactic factors and growth factors involved in the inflammatory responses,which responses exist in all stages of development of atherosclerosis.Studies have shown that proinflammatory factors can inhibit macrophage reverse cholesterol transport and promote the foam cells.Therefore.searching active compounds of anti-inflammatory effect in macrophage,offer a new way for the treatment of AS.To systematically explore its anti-atherosclerotic effects in vitro and in vivo,we investigated the mechanism of the asperlin suppress inflammation and foam cells formation in RAW264.7 cells,and the pharmaceutical effect in ApoE-/-mice.This article adopted the LPS-induced foam cells model to investigate the inhibitory effect of asperlin on foam cells formation and its molecular mechanism.Oil red O staining,total triglyceride(TG)and cholesterol(TC)specific kits were performed to assess asperlin's influence on lipid accumulation;The fluorescence-labeled assay were tested to analyze effect of asperlin on cholesterol flux;Real-time quantitative PCR detected related genes mRNA expression.Western blot detected related protein.Subsequently,we explored the relationship of asperlin with inflammation in macrophage.ELISA kits tested proinflammatory factor by treatment with asperlin in LPS-provoked macrophages;Real-time quantitative PCR detected mRNA expressions related to inflammatory factor.The results revealed treatment of asperlin at the dose of 10 ?mol/L significantly inhibited triglyceride accumulation and regulated high-density lipoprotein(HDL)mediated cholesterol flux which proved the effect on inhibiting LPS-evoked foam cells formation.RT-PCR and western blot results showed that the asperlin remarkably regulated the cholesterol flux related gene mRNA levels and protein levels.Above results indicated that asperlin inhibit foam cells formation.Supplementation with asperlin also suppressed LPS-elicited production of pro-inflammatory factors in RAW264.7 macrophages,decreased the expression levels of iNOS,IL-1? and TNF?,and increased the expression of IL-10 and IL-4,indicating remarkable shift in M1/M2 macrophages polarization.In vivo experiments,We fed ApoE-/-mice with a cholesterol-rich diet for 12 weeks to establish an atherosclerotic animal model and investigate the atheropreventive effect of asperlin in vivo.At 12th week,the vascular lumen diameter was measured by high frequency ultrasound.The animals were killed and collected blood after 12 weeks of treatment.We also evaluated the serum concentration of lipid as well as serum levels of pro-inflammatory factors and the lesion size was determined in the valve area of the aortic root to explore effect and mechanism of asperlin prevents atherosclerosis in vivo.The results showed that oral administration of asperlin for 12 weeks remarkably suppressed atherosclerotic plaque formation in aorta,as revealed by reduced aortic dilatation and decreased atherosclerotic lesion area.Asperlin also decreased serum levels of pro-inflammatory factors but showed little impact on blood lipids in ApoE-/-atherosclerotic mice.These results revealed that asperlin suppress atherosclerosis by inhibiting inflammation rather than decreasing the level of total cholesterol in ApoE-/-mice.In conclusion,we first proved that asperlin,which is a marine-derived natural product,can inhibit LPS-evoked foam cell formation by suppressing macophage inflammation.Asperlin is adequate to prevent atherosclerosis in vivo.It may exert atheroprotective function through suppressing inflammation rather than ameliorating dyslipidemia.The data provided an experimental reference of asperlin on prevention and treatment on atherosclerosis. |
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