Berberine ameliorates vascular dysfunction by a global modulation of lncRNA and mRNA expression profiles in hypertensive mouse aortae.

Na Tan,Limei Liu,Yan Zhang,Wenwen Han,Yi Zong,Li Li,Yi Zhang,Yu Huang

doi:10.1371/journal.pone.0247621

Abstract

The current study investigated the mechanism underlying the therapeutic effects of berberine in the vasculature in hypertension. Angiotensin II (Ang II)-loaded osmotic pumps were implanted in C57BL/6J mice with or without berberine administration. Mouse aortae were suspended in myograph for force measurement. Microarray technology were performed to analyze expression profiles of lncRNAs and mRNAs in the aortae. These dysregulated expressions were then validated by qRT-PCR. LncRNA-mRNA co-expression network was constructed to reveal the specific relationships. Ang Ⅱ resulted in a significant increase in the blood pressure of mice, which was suppressed by berberine. The impaired endothelium-dependent aortic relaxation was restored in hypertensive mice. Microarray data revealed that 578 lncRNAs and 554 mRNAs were up-regulated, while 320 lncRNAs and 377 mRNAs were down-regulated in the aortae by Ang Ⅱ; both were reversed by berberine treatment. qRT-PCR validation results of differentially expressed genes (14 lncRNAs and 6 mRNAs) were completely consistent with the microarray data. GO analysis showed that these verified differentially expressed genes were significantly enriched in terms of "cellular process", "biological regulation" and "regulation of biological process", whilst KEGG analysis identified vascular function-related pathways including cAMP signaling pathway, cGMP-PKG signaling pathway, and calcium signaling pathway etc. Importantly, we observed that lncRNA ENSMUST00000144849, ENSMUST00000155383, and AK041185 were majorly expressed in endothelial cells. The present results suggest that the five lncRNAs ENSMUST00000144849, NR_028422, ENSMUST00000155383, AK041185, and uc.335+ might serve critical regulatory roles in hypertensive vasculature by targeting pivotal mRNAs and subsequently affecting vascular function-related pathways. Moreover, these lncRNAs were modulated by berberine, therefore providing the novel potential therapeutic targets of berberine in hypertension. Furthermore, lncRNA ENSMUST00000144849, ENSMUST00000155383, and AK041185 might be involved in the preservation of vascular endothelial cell function.

Highlights

Hypertension is a leading common risk factor for worldwide morbidity and mortality from cardiovascular diseases [1,2,3], which is associated with the alterations in the function and structure of resistance and conduit arteries
Microarray data revealed that 578 Long noncoding RNAs (lncRNAs) and 554 mRNAs were up-regulated, while 320 lncRNAs and 377 mRNAs were down-regulated in the aortae by Angiotensin II (Ang II); both were reversed by berberine treatment. quantitative real-time polymerase chain reaction (qRT-PCR) validation results of differentially expressed genes (14 lncRNAs and 6 mRNAs) were completely consistent with the microarray data
Gene ontology (GO) analysis showed that these verified differentially expressed genes were significantly enriched in terms of “cellular process”, “biological regulation” and “regulation of biological process”, whilst Kyoto Encyclopedia of genes and genomes (KEGG) analysis identified vascular function-related pathways including cAMP signaling pathway, cGMP-PKG signaling pathway, and calcium signaling pathway etc

Summary

Introduction

Hypertension is a leading common risk factor for worldwide morbidity and mortality from cardiovascular diseases [1,2,3], which is associated with the alterations in the function and structure of resistance and conduit arteries. Multiple beneficial influences of berberine on cardiovascular system have been reported, including anti-hyperglycemic, antioxidative, cholesterol-lowering, and cardiac protective effects [8,9,10] Both animal studies and clinical experiments indicate that berberine exerts protective effects on vascular endothelial function. Berberine attenuates carotid arterial endothelium-dependent contractions by inhibiting ER stress [11] and ameliorates aortic endothelial dysfunction [12] in spontaneously hypertensive rats (SHR), postponing the progress of hypertension. It improves flow-mediated vasodilation (FMD) possibly via reducing circulating endothelial microparticles in human [13]. Combination therapy with berberine and trimetazidine significantly increases FMD in patients with coronary heart disease and primary hypertension [14]

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Results

Discussion

Conclusion