Effects of chrysin (5,7-dihydroxyflavone) on vascular remodeling in hypoxia-induced pulmonary hypertension in rats

Chinese Medicine

Jie-Ren Yang¹  Shu Li²  Xiang-Ming Wang³  Xian-Wei Li¹ 

[1] Department of Pharmacology, Wannan Medical College, Anhui 241002, China;Department of Pathophysiology, Wannan Medical College, Anhui 241002, China;Department of Pathology, Yijishan Hospital, Wannan Medical College, Anhui 241002, China

关键词: Collagen;    Reactive oxygen species;    NOX4;    Pulmonary hypertension;    Chrysin;   

Others  :  1135249
DOI  :  10.1186/s13020-015-0032-2

 received in 2014-07-25, accepted in 2015-02-09,  发布年份 2015

【 摘 要 】

Background

Chrysin (5,7-dihydroxyflavone) inhibits platelet-derived growth factor-induced vascular smooth muscle cell proliferation and arterial intima hyperplasia. This study aims to investigate the effects of chrysin on rat pulmonary vascular remodeling in hypoxia-induced pulmonary hypertension (PH).

Methods

Sprague–Dawley rats were continuously exposed to 10% O₂ for 4 weeks to induce PH. The effect of chrysin (50 or 100 mg/kg/d, subcutaneous) on vascular remodeling was investigated in hypoxia-induced PH model. At the end of the experiments, the indexes for pulmonary vascular remodeling and right ventricle hypertrophy were measured by vascular medial wall thickness and the ratio of right ventricle to (left ventricle plus septum). The expressions of NOX4, collagen I, and collagen III were analyzed by immunohistochemistry, real-time PCR, or western blotting. The proliferation of cultured pulmonary artery smooth muscle cells (PASMCs) was determined by BrdU incorporation and flow cytometry. The levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were also determined by thiobarbituric acid reactive substances assay and 2′7′-dichlorofluorescein diacetate method.

Results

Chrysin treatment for 4 weeks significantly attenuated pulmonary vascular remodeling and improved collagen accumulation and down-regulated collagen I and collagen III expressions, accompanied by downregulation of NOX4 expression in the pulmonary artery (P = 0.012 for 50 mg/kg/d, P < 0.001 for 100 mg/kg/d) and lung tissue (P = 0.026, P < 0.001). In vitro, chrysin (1, 10, and 100 μM) remarkably attenuated PASMC proliferation (P = 0.021 for 1 μM, P < 0.001 for 10 μM, and P < 0.001 for 100 μM), collagen I expression (P = 0.035, P < 0.001, and P < 0.001), and collagen III expression (P = 0.027, P < 0.001, and P < 0.001) induced by hypoxia, and these inhibitory effects of chrysin were accompanied by inhibition of NOX4 expression (P = 0.019, P < 0.001, and P < 0.001), ROS production (P = 0.038, P < 0.001, and P < 0.001), and MDA generation (P = 0.024, P < 0.001, and P < 0.001).

Conclusions

This study demonstrated that chrysin treatment in hypoxia-induced PH in rats reversed the hypoxia-induced (1) elevations of NOX4 expression, (2) productions of ROS and MDA, (3) proliferation of PASMC, and (4) accumulation of collagen.

【 授权许可】

   
2015 Li et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Crossno JT Jr, Garat CV, Reusch JE, Morris KG, Dempsey EC, McMurtry IF, et al.: Rosiglitazone attenuates hypoxia-induced pulmonary arterial remodeling. Am J Physiol Lung Cell Mol Physiol 2007, 292:L885-97.
• [2]Dahal BK, Cornitescu T, Tretyn A, Pullamsetti SS, Kosanovic D, Dumitrascu R, et al.: Role of epidermal growth factor inhibition in experimental pulmonary hypertension. Am J Respir Crit Care Med 2010, 181:158-67.
• [3]Sakao S, Tatsumi K: Vascular remodeling in pulmonary arterial hypertension: multiple cancer-like pathways and possible treatment modalities. Int J Cardiol 2011, 147:4-12.
• [4]Hoeper MM: Pulmonary hypertension in collagen vascular disease. Eur Respir J 2002, 19:571-6.
• [5]Tanoue LT: Pulmonary hypertension in the collagen vascular diseases. Semin Respir Crit Care Med 2003, 24:287-96.
• [6]Li N, Dai DZ, Dai Y: CPU86017 and its isomers improve hypoxic pulmonary hypertension by attenuating increased ETA receptor expression and extracellular matrix accumulation. Naunyn Schmiedebergs Arch Pharmacol 2008, 378:541-52.
• [7]Sahara M, Takahashi T, Imai Y, Nakajima T, Yao A, Morita T, et al.: New insights in the treatment strategy for pulmonary arterial hypertension. Cardiovasc Drugs Ther 2006, 20:377-86.
• [8]Demarco VG, Whaley-Connell AT, Sowers JR, Habibi J, Dellsperger KC: Contribution of oxidative stress to pulmonary arterial hypertension. World J Cardiol 2010, 2:316-24.
• [9]Tabima DM, Frizzell S, Gladwin MT: Reactive oxygen and nitrogen species in pulmonary hypertension. Free Radic Biol Med 2012, 52:1970-86.
• [10]Stenmark KR, Fagan KA, Frid MG: Hypoxia-induced pulmonary vascular remodeling: cellular and molecular mechanisms. Circ Res 2006, 99:675-91.
• [11]Brennan LA, Steinhorn RH, Wedgwood S, Mata-Greenwood E, RoarkE A, Russell JA, et al.: Increased superoxide generation is associated with pulmonary hypertension in fetal lambs: a role for NADPH oxidase. Circ Res 2003, 92:683-91.
• [12]Wedgwood S, Dettman RW, Black SM: ET-1 stimulates pulmonary arterial smooth muscle cell proliferation via induction of reactive oxygen species. Am J Physiol Lung Cell Mol Physiol 2001, 281:L1058-67.
• [13]Liu JQ, Sham JS, Shimoda LA, Kuppusamy P, Sylvester JT: Hypoxic constriction and reactive oxygen species in porcine distal pulmonary arteries. Am J Physiol Lung Cell Mol Physiol 2003, 285:L322-33.
• [14]Hoshikawa Y, Ono S, Suzuki S, Tanita T, Chida M, Song C, et al.: Generation of oxidative stress contributes to the development of pulmonary hypertension induced by hypoxia. J Appl Physiol 2001, 90:1299-306.
• [15]Mittal M, Roth M, Ko¨nig P, Hofmann S, Dony E, Goyal P, et al.: Hypoxia-dependent regulation of non-phagocytic NADPH oxidase subunit NOX4 in the pulmonary vasculature. Circ Res 2007, 101:258-67.
• [16]Ismail S, Sturrock A, Wu P, Cahill B, Norman K, Huecksteadt T, et al.: NOX4 mediates hypoxia-induced proliferation of human pulmonary artery smooth muscle cells: the role of autocrine production of transforming growth factor-{beta}1 and insulin-like growth factor binding protein-3. Am J Physiol Lung Cell Mol Physiol 2009, 296:L489-99.
• [17]Green DE, Kang BY, Murphy TC, Hart CM: Peroxisome proliferator-activated receptor gamma (PPARγ) regulates thrombospondin-1 and Nox4 expression in hypoxia-induced human pulmonary artery smooth muscle cell proliferation. Pulm Circ 2012, 2:483-91.
• [18]Nisbet RE, Bland JM, Kleinhenz DJ, Mitchell PO, Walp ER, Sutliff RL, et al.: Rosiglitazone attenuates chronic hypoxia-induced pulmonary hypertension in a mouse model. Am J Respir Cell Mol Biol 2010, 42:482-90.
• [19]Williams CA, Harborne JB, Newman M, Greenham J, Eagles J: Chrysin and other leaf exudate flavonoids in the genus Pelargonium. Phytochemistry 1997, 46:1349-53.
• [20]Rapta P, Misík V, Stasko A, Vrábel I: Redox intermediates of flavonoids and caffeic acid esters from propolis: an EPR spectroscopy and cyclic voltammetry study. Free Radic Biol Med 1995, 18:901-8.
• [21]Cho H, Yun CW, Park WK, Kong JY, Kim KS, Park Y, et al.: Modulation of the activity of pro-inflammatory enzymes, COX-2 and iNOS, by chrysin derivatives. Pharmacol Res 2004, 49:37-43.
• [22]Zhang T, Chen X, Qu L, Wu J, Cui R, Zhao Y: Chrysin and its phosphate ester inhibit cell proliferation and induce apoptosis in Hela cells. Bioorg Med Chem 2004, 12:6097-105.
• [23]Chaudhuri S, Banerjee A, Basu K, Sengupta B, Sengupta PK: Interaction of flavonoids with red blood cell membrane lipids and proteins: antioxidant and antihemolytic effects. Int J Biol Macromol 2007, 41:42-8.
• [24]Pushpavalli G, Kalaiarasi P, Veeramani C, Pugalendi KV: Effect of chrysin on hepatoprotective and antioxidant status in D-galactosamine-induced hepatitis in rats. Eur J Pharmacol 2010, 631:36-41.
• [25]Villar IC, Jiménez R, Galisteo M, Garcia-Saura MF, Zarzuelo A, Duarte J: Effects of chronic chrysin treatment in spontaneously hypertensive rats. Planta Med 2002, 68:847-50.
• [26]Villar IC, Galisteo M, Vera R, O’Valle F, García-Saura MF, Zarzuelo A, et al.: Effects of the dietary flavonoid chrysin in isolated rat mesenteric vascular bed. J Vasc Res 2004, 41:509-16.
• [27]Villar IC, Vera R, Galisteo M, O’Valle F, Romero M, Zarzuelo A, et al.: Endothelial nitric oxide production stimulated by the bioflavonoid chrysin in rat isolated aorta. Planta Med 2005, 71:829-34.
• [28]Lo HM, Wu MW, Pan SL, Peng CY, Wu PH, Wu WB: Chrysin restores PDGF-induced inhibition on protein tyrosine phosphatase and reduces PDGF signaling incultured VSMCs. J Nutr Biochem 2012, 23:667-78.
• [29]Li XW, Hu CP, Wu WH, Zhang WF, Zou XZ, Li YJ: Inhibitory effect of calcitonin gene-related peptide on hypoxia-induced rat pulmonary artery smooth muscle cells proliferation: role of ERK1/2 and p27. Eur J Pharmacol 2012, 679:117-26.
• [30]Li XW, Du J, Hu GY, Hu CP, Li D, Li YJ, et al.: Fluorofenidone attenuates vascular remodeling in hypoxia-induced pulmonary hypertension of rats. Can J Physiol Pharmacol 2014, 92:58-69.
• [31]Janero DR: Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 1990, 9:515-40.
• [32]Fredenburgh LE, Liang OD, Macias AA, Polte TR, Liu X, Riascos DF, et al.: Absence of cyclooxygenase-2 exacerbates hypoxia-induced pulmonary hypertension and enhances contractility of vascular smooth muscle cells. Circulation 2008, 117:2114-22.
• [33]Li J, Zhang P, Zhang QY, Zhang SM, Guo HT, Bi H, et al.: Effects of U50, 488H on hypoxia pulmonary hypertension and its underlying mechanism. Vascul Pharmacol 2009, 51:72-7.
• [34]Gong LM, Du JB, Shi L, Shi Y, Tang CS: Effects of endogenous carbon monoxide on collagen synthesis in pulmonary artery in rats under hypoxia. Life Sci 2004, 74:1225-41.
• [35]Fike CD, Aschner JL, Slaughter JC, Kaplowitz MR, Zhang Y, Pfister SL: Pulmonary arterial responses to reactive oxygen species are altered in newborn piglets with chronic hypoxia-induced pulmonary hypertension. Pediatr Res 2011, 70:136-41.
• [36]Wolin MS, Ahmad M, Gupte SA: Oxidant and redox signaling in vascular oxygen sensing mechanisms: basic concepts, current controversies, and potential importance of cytosolic NADPH. Am J Physiol Lung Cell Mol Physiol 2005, 289:L159-73.
• [37]Liu JQ, Zelko IN, Erbynn EM, Sham JS, Folz RJ: Hypoxic pulmonary hypertension: role of superoxide and NADPH oxidase (gp91phox). Am J Physiol Lung Cell Mol Physiol 2006, 290:L2-10.
• [38]Sorescu D, Weiss D, Lassegue B, Clempus RE, Szocs K, Sorescu GP, et al.: Superoxide production and expression of nox family proteins in human atherosclerosis. Circulation 2002, 105:1429-35.
• [39]Szocs K, Lassegue B, Sorescu D, Hilenski LL, Valppu L, Couse TL, et al.: Upregulation of Nox-based NAD (P) H oxidases in restenosis after carotid injury. Arterioscler Thromb Vasc Biol 2002, 22:21-7.