Abstract
MicroRNAs (miRNAs) are a class of small noncoding RNAs, producing transcripts of about 22 nucleotides in length. miRNAs usually function as antisense regulators of other RNAs by degrading their targets. Recently, miRNAs have emerged as interesting new drug targets due to their regulatory role in essential biological processes. Salvianolic acid B (SalB) is one of the major pharmacologically active ingredients of Salvia miltiorrhiza, a traditional oriental medicine for treatment of cardiovascular disorders. In this study, we determined whether miRNAs play a role in regulation of various gene expression responses to SalB in human umbilical vein endothelial cells (HUVECs). We used the microarray approach to evaluate levels of both miRNA and mRNA, and found that 171 miRNAs were differentially expressed in SalB-treated HUVECs. We additionally identified 848 messenger RNAs (mRNAs) that are anti-correlated with the miRNAs expression. The Gene Ontology (GO) term enrichment was analyzed for identification of biological processes of target genes affected by differential expression of miRNA. Among 848 genes investigated, cardiovascular diseaserelated genes were selected in SalB-treated HUVECs. These results suggest that SalB may modulate miRNA and their target gene expression in order to exert vascular protective effects in human endothelial cells.
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Filipowicz, W., Bhattacharyya, S. N. & Sonenberg, N. Mechanisms of post-transcriptional regulation by micro RNAs: are the answers in sight? Nat Rev Genet 9:102–114 (2008).
Urbich, C., Kuehbacher, A. & Dimmeler, S. Role of microRNAs in vascular diseases, inflammation, and angiogenesis. Cardiovasc Res 79:581–588 (2008).
Wang, S. X., Hu, L. M., Gao, X. M., Guo, H. & Fan, G. W. Anti-inflammatory activity of salvianolic acid B in microglia contributes to its neuroprotective effect. Neurochem Res 35:1029–1037 (2010).
Zhou, Z., Liu, Y., Miao, A. D. & Wang, S. Q. Salvianolic acid B attenuates plasminogen activator inhibitor type 1 production in TNF-alpha treated human umbilical vein endothelial cells. J Cell Biochem 96: 109–116 (2005).
Chen, Y. L. et al. Salvianolic acid B attenuates cyclooxygenase-2 expression in vitro in LPS-treated human aortic smooth muscle cells and in vivo in the apolipoprotein-E-deficient mouse aorta. J Cell Biochem 98: 618–631 (2006).
Liu, C. S., Chen, N. H. & Zhang, J. T. Protection of PC12 cells from hydrogen peroxide-induced cytotoxicity by salvianolic acid B, a new compound isolated from Radix Salviae miltiorrhizae. Phytomedicine 14: 492–497 (2007).
Lam, F. F., Yeung, J. H., Kwan, Y. W., Chan, K. M. & Or, P. M. Salvianolic acid B, an aqueous component of danshen (Salvia miltiorrhiza), relaxes rat coronary artery by inhibition of calcium channels. Eur J Pharmacol 553:240–245 (2006).
Zhou, Y., Gu, J. & Xu, L. M. Effect and mechanism of salvianolic acid B in attenuating elevated portal pressure in a rat model of portal hypertension induced by endothelin-1. Zhong Xi Yi Jie He Xue Bao 5:61–64 (2007).
Lin, S. J. et al. Salvianolic acid B attenuates MMP-2 and MMP-9 expression in vivo in apolipoprotein-Edeficient mouse aorta and in vitro in LPS-treated human aortic smooth muscle cells. J Cell Biochem 100:372–384 (2007).
Chen, Y. H. et al. Salvianolic acid B attenuates VCAM-1 and ICAM-1 expression in TNF-alpha-treated human aortic endothelial cells. J Cell Biochem 82:512–521 (2001).
Yan, Q., Yao-Cheng, R., Li, Z., Tie-Jun, L. & Wei-Dong, Z. VEGF induced hyperpermeability in bovine aortic endothelial cell and inhibitory effect of salvianolic acid B. Acta Pharmacologica Sinica 22:117–120 (2001).
Ding, M., Ye, T. X., Zhao, G. R., Yuan, Y. J. & Guo, Z. X. Aqueous extract of Salvia miltiorrhiza attenuates increased endothelial permeability induced by tumor necrosis factor-alpha. Int Immunopharmacol 5:1641–1651 (2005).
Wu, H. L. et al. Salvianolic acid B protects human endothelial cells from oxidative stress damage: a possible protective role of glucose-regulated protein 78 induction. Cardiovasc Res 81:148–158 (2009).
Zhang, J., Zhao, G., Liu, J. & Ji, X. Protection of salvianolic acid B for human endothelial cells against hydrogen peroxide-induced oxidative damage. Transactions of Tianjin University 15:434–439 (2009).
Chen, S. C., Lin, Y. L., Huang, B., Wang, D. L. & Cheng, J. J. Salvianolic acid B suppresses IFN-gamma-induced JAK/STAT1 activation in endothelial cells. Thromb Res 128:560–564 (2011).
Zhao, J. F. et al. Effect of salvianolic acid B on Smad3 expression in hepatic stellate cells. Hepatobiliary Pancreat Dis Int 3:102–105 (2004).
Dennis, G., Jr. et al. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol 4:P3 (2003).
Sumpio, B. E., Riley, J. T. & Dardik, A. Cells in focus: endothelial cell. Int J Biochem Cell Biol 34:1508–1512 (2002).
Wang, Q. L., Wu, Q., Tao, Y. Y., Liu, C. H. & El-Nezami, H. Salvianolic acid B modulates the expression of drug-metabolizing enzymes in HepG2 cells. Hepatobiliary Pancreat Dis Int 10:502–508 (2011).
Wu, Y. P. et al. Salvianolic acid B inhibits platelet adhesion under conditions of flow by a mechanism involving the collagen receptor alpha2beta1. Thromb Res 123:298–305 (2008).
Xu, L. L. et al. Cardio-Protection of Salvianolic Acid B through Inhibition of Apoptosis Network. PLoS One 6:e24036 (2011).
Yang, X. Y. et al. Salvianolic acid A protects against vascular endothelial dysfunction in high-fat diet fed and streptozotocin-induced diabetic rats. J Asian Nat Prod Res 13:884–894 (2011).
Ness, S. A. Microarray analysis: basic strategies for successful experiments. Mol Biotechnol 36:205–219 (2007).
Lim, L. P. et al. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature 433:769–773 (2005).
Lorenzen, J., Kumarswamy, R., Dangwal, S. & Thum, T. MicroRNAs in diabetes and diabetes-associated complications. RNA Biol 9:820–827(2012).
Lee, S. E. et al. MicroRNA and gene expression analysis of melatonin-exposed human breast cancer cell lines indicating involvement of the anticancer effect. J Pineal Res 51:345–352 (2011).
Zernecke, A. MicroRNAs in the regulation of immune cell functions — implications for atherosclerotic vascular disease. Thromb Haemost 107:626–633 (2012).
Zampetaki, A. & Mayr, M. MicroRNAs in vascular and metabolic disease. Circ Res 110:508–522 (2012).
Harris, T. A., Yamakuchi, M., Ferlito, M., Mendell, J. T. & Lowenstein, C. J. MicroRNA-126 regulates endothelial expression of vascular cell adhesion molecule 1. Proc Natl Acad Sci USA 105:1516–1521 (2008).
Lee, S. E. et al. Uncaria rhynchophylla induces heme oxygenase-1 as a cytoprotective effect in RAW 264.7 macrophages. Mol. Cell. Toxicol. 6:33–40 (2010).
Kim, J. H. Cardiovascular Diseases and Panax ginseng: A Review on Molecular Mechanisms and Medical Applications. J. Ginseng Res. 36:16–26 (2012).
Choi, K. S. et al. Inhibition of Hydrogen Sulfide-induced Angiogenesis and Inflammation in Vascular Endothelial Cells: Potential Mechanisms of Gastric Cancer Prevention by Korean Red Ginseng. J. Ginseng Res. 36:135–145 (2012).
Yang, F. G. et al. Effects of salvianolic acid B on cardiovascular endothelial cells and platelet activation in a rabbit model of ischemia-reperfusion. Zhong Xi Yi Jie He Xue Bao 6:1250–1254 (2008).
Arola-Arnal, A. & Blade, C. Proanthocyanidins modulate microRNA expression in human HepG2 cells. PLoS One 6:e25982 (2011).
Joven, J. et al. Plant-derived polyphenols regulate expression of miRNA paralogs miR-103/107 and miR-122 and prevent diet-induced fatty liver disease in hyperlipidemic mice. Biochim Biophys Acta 1820:894–899 (2012).
Milenkovic, D. et al. Modulation of miRNA expression by dietary polyphenols in apoE deficient mice: a new mechanism of the action of polyphenols. PLoS One 7:e29837 (2012).
Wang, H., Bian, S. & Yang, C. S. Green tea polyphenol EGCG suppresses lung cancer cell growth through upregulating miR-210 expression caused by stabilizing HIF-1alpha. Carcinogenesis 32:1881–1889 (2011).
Wen, X. Y. et al. Ellagitannin (BJA3121), an anti-proliferative natural polyphenol compound, can regulate the expression of MiRNAs in HepG2 cancer cells. Phytother Res 23:778–784 (2009).
Yang, H. et al. Expression profile analysis of human umbilical vein endothelial cells treated with salvianolic acid B from Salvia miltiorrhiza. BioChip J. 5:47–55 (2011).
Lee, S. E. et al. Methylglyoxal-mediated alteration of gene expression in human endothelial cells. BioChip J. 5:220–228 (2011).
Yang, H. et al. Up-regulation of Heme Oxygenase-1 by Korean Red Ginseng Water Extract as a Cytoprotective Effect in Human Endothelial Cells. J. Ginseng Res. 35:352–359 (2011).
Lee, S. E. et al. Induction of Heme Oxygenase-1 Inhibits Cell Death in Crotonaldehyde-Stimulated HepG2 Cells via the PKC-delta-p38-Nrf2 Pathway. PLoS One 7:e41676 (2012).
Lee, S. E. et al. Genome-wide profiling in melatoninexposed human breast cancer cell lines identifies differentially methylated genes involved in the anticancer effect of melatonin. J Pineal Res 54:80–88 (2012).
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Yang, H., Lee, S.E., Kim, GD. et al. An integrated analysis of microRNA and mRNA expression in salvianolic acid B-treated human umbilical vein endothelial cells. Mol. Cell. Toxicol. 9, 1–7 (2013). https://doi.org/10.1007/s13273-013-0001-8
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DOI: https://doi.org/10.1007/s13273-013-0001-8