Vaccarin attenuates the human EA.hy926 endothelial cell oxidative stress injury through inhibition of Notch signaling

Xie,Fengshan; Cai,Weiwei; Liu,Yanling; Li,Yue; Du,Bin; Feng,Lei; Qiu,Liying

doi:10.3892/ijmm.2014.1977

Vaccarin attenuates the human EA.hy926 endothelial cell oxidative stress injury through inhibition of Notch signaling

Authors:
- Fengshan Xie
- Weiwei Cai
- Yanling Liu
- Yue Li
- Bin Du
- Lei Feng
- Liying Qiu
View Affiliations

Affiliations: Laboratory of Natural Medicine, School of Pharmaceutical Science, Jiangnan University, Wuxi, Jiangsu 214122, P.R. China, Wuxi Medical School, Jiangnan University, Wuxi, Jiangsu 214122, P.R. China
Published online on: October 23, 2014 https://doi.org/10.3892/ijmm.2014.1977
Pages: 135-142

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Endothelial cell injury is an essential component of atherosclerosis and hypertension. Atherosclerosis and other macrovascular diseases are the most common complications of diabetes. Vaccarin is a major flavonoid glycoside in Vaccariae semen, and is expected to be useful in the treatment of vascular diseases. The aim of the present study was to evaluate the possible effects of vaccarin in human umbilical vein endothelial cells (EA.hy926) induced by hydrogen peroxide (H2O2) and its underlying mechanism in the prevention and treatment of H2O2 injury. In this study, the EA.hy926 cells were exposed to 250, 500 and 1000 µM H2O2 for 2 and 4 h in the absence or presence of vaccarin, and the cell injury induced by H2O2 was examined via SRB. Cell migratory ability, lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) levels and decreasing superoxide dismutase (SOD) activity were evaluated by the wound healing assay and corresponding assay kits. Cell apoptosis was detected by flow cytometry with Annexin V‑fluorescein isothiocyanate/propidium iodide Apoptosis Detection kit and Hoechst staining. Furthermore, western blot detected the protein expressions of Notch1, Hes1 and caspase‑3. Following treatment with H2O2, it was found that H2O2 stimulated cell injury in a dose‑dependent manner, including reducing cell viability and cell migratory ability, increasing LDH leakage and MDA levels, and decreasing SOD activity. H2O2 further accelerated cell apoptosis via activation of Notch1 and the downstream molecule Hes1. Preincubation with vaccarin was found to protect EA.hy926 cells from H2O2‑induced cell oxidative stress injury, which promoted cell viability and cell migratory ability, inhibited the level of LDH and MDA, but enhanced the activity of SOD. In particular, in addition to downregulation Notch signaling, vaccarin treatments also downregulated caspase‑3, a cell apoptotic pathway‑related protein. These findings indicated that vaccarin may be able to selectively protect vascular endothelium from dysfunction induced by H2O2.

View Figures

View References

1	Beckman JA, Creager MA and Libby P: Diabetes and atherosclerosis: epidemiology, pathophysiology, and management. JAMA. 287:2570–2581. 2002. View Article : Google Scholar : PubMed/NCBI
2	Malakul W, Thirawarapan S, Suvitayavat W and Woodman OL: Type 1 diabetes and hypercholesterolaemia reveal the contribution of endothelium-derived hyperpolarizing factor to endothelium-dependent relaxation of the rat aorta. Clin Exp Pharmacol Physiol. 35:192–200. 2008.
3	McNulty PH, Tulli MA, Robertson BJ, et al: Effect of simulated postprandial hyperglycemia on coronary blood flow in cardiac transplant recipients. Am J Physiol Heart Circ Physiol. 293:H103–H108. 2007. View Article : Google Scholar : PubMed/NCBI
4	Vita JA: Endothelial function and clinical outcome. Heart. 91:1278–1279. 2005. View Article : Google Scholar : PubMed/NCBI
5	Schulz E, Dopheide J, Schuhmacher S, et al: Suppression of the JNK pathway by induction of a metabolic stress response prevents vascular injury and dysfunction. Circulation. 118:1347–1357. 2008. View Article : Google Scholar : PubMed/NCBI
6	Yada T, Shimokawa H, Hiramatsu O, et al: Hydrogen peroxide, an endogenous endothelium-derived hyperpolarizing factor, plays an important role in coronary autoregulation in vivo. Circulation. 107:1040–1045. 2003. View Article : Google Scholar : PubMed/NCBI
7	West XZ, Malinin NL, Merkulova AA, et al: Oxidative stress induces angiogenesis by activating TLR2 with novel endogenous ligands. Nature. 467:972–976. 2010. View Article : Google Scholar : PubMed/NCBI
8	Chen CA, Wang TY, Varadharaj S, et al: S-glutathionylation uncouples eNOS and regulates its cellular and vascular function. Nature. 468:1115–1118. 2010. View Article : Google Scholar : PubMed/NCBI
9	Wu XG and Li L: Rosiglitazone suppresses lipopolysaccharide-induced matrix metalloproteinase-2 activity in rat aortic endothelial cells via Ras-MEK1/2 signaling. Int J Cardiol. 158:54–58. 2012. View Article : Google Scholar
10	Zanchetti A, Hennig M, Hollweck R, et al: Baseline values but not treatment-induced changes in carotid intima-media thickness predict incident cardiovascular events in treated hypertensive patients: findings in the European Lacidipine Study on Atherosclerosis (ELSA). Circulation. 120:1084–1090. 2009. View Article : Google Scholar : PubMed/NCBI
11	Yang Y, Duan WX, Liang ZX, et al: Curcumin attenuates endothelial cell oxidative stress injury through Notch signaling inhibition. Cell Signal. 25:615–629. 2013. View Article : Google Scholar
12	Rosenbaum MA, Miyazaki K and Graham LM: Hypercholesterolemia and oxidative stress inhibit endothelial cell healing after arterial injury. J Vasc Surg. 55:489–496. 2012. View Article : Google Scholar :
13	Cook KM and Figg WD: Angiogenesis inhibitors: Current strategies and future prospects. CA Cancer J Clin. 60:222–243. 2010. View Article : Google Scholar : PubMed/NCBI
14	Ji X, Wang Z, Geamanu A, Sarkar FH and Gupta SV: Inhibition of cell growth and induction of apoptosis in non-small cell lung cancer cells by delta-tocotrienol is associated with notch-1 down-regulation. J Cell Biochem. 112:2773–2783. 2011. View Article : Google Scholar : PubMed/NCBI
15	Samon JB, Champhekar A, Minter LM, et al: Notch1 and TGFb1 cooperatively regulate Foxp3 expression and the maintenance of peripheral regulatory T cells. Blood. 112:1813–1821. 2008. View Article : Google Scholar : PubMed/NCBI
16	Maciej J and Krzysztof S: Notch: A new player in MS mechanisms. J Neuroimmunol. 218:3–11. 2010. View Article : Google Scholar
17	Androutsellis-Theotokis A, Leker RR, Soldner F, et al: Notch signalling regulates stem cell numbers in vitro and in vivo. Nature. 442:823–826. 2006. View Article : Google Scholar : PubMed/NCBI
18	McCright B: Notch signaling in kidney development. Curr Opin Nephrol Hypertens. 12:5–10. 2003. View Article : Google Scholar
19	Archana VB, Karl DP, Pao LC, Yoon YS and Michael ED: Oxidative stress-induced Notch1 signaling promotes cardiogenic gene expression in mesenchymal stem cells. Stem Cell Res Ther. 4:2–15. 2013.
20	Campos J, Schmeda-Hirschmann G, Leiva E, et al: Lemon grass (Cymbopogon citratus (D.C) Stapf) polyphenols protect human umbilical vein endothelial cell (HUVECs) from oxidative damage induced by high glucose, hydrogen peroxide and oxidised low-density lipoprotein. Food Chem. 151:175–181. 2014. View Article : Google Scholar : PubMed/NCBI
21	Jin RG, Wang QJ and Yan TH: Protective effects of salidroside on human umbilical vein endothelial cells. Pharmacology and Clinics of Chinese Materia Medical. 28:41–44. 2012.
22	China Pharmacopoeia Committee. Chinese Pharmacopoeia: The 2010 edition. China Medical Science Press; Beijing, China: pp. 49–50. 2010
23	Sang SM, Lao A and Chen ZL: Chemistry and bioactivity of the seeds of Vaccaria segetalis. Orient Foods Herbs. 21:279–291. 2000.
24	Li F and Liang JY: Research progress of Vaccaria segetalis. Straits Pharm J. 3:1–5. 2007.
25	Koike K, Jia ZH and Nikaido T: Triterpenoid saponins from Vaccaria segetalis. Phytochemistry. 7:13431998. View Article : Google Scholar
26	Sang SM, Lao AN and Wang HC: A phenylpropanoid glycoside from Vaccaria segetalis. Phytochemistry. 48:569–571. 1998. View Article : Google Scholar
27	Sang S, Lao A and Wang H: Triterpenoid saponin from Vaccaria segetalis. J Asian Nat Prod Res. 1:199–205. 1999. View Article : Google Scholar
28	Yun YS, Morita H, Takeya K, et al: Cyclic peptides from higher plants. 34. segetalins G and H, structures and estrogen-like activity of cyclic pentapeptides from Vaccaria segetalis. J Nat Prod. 60:216–218. 1997. View Article : Google Scholar : PubMed/NCBI
29	Morita H, Young SY, Takeya K, et al: A cyclic heptapeptide from Vaccaria segetalis. Phytochemistry. 42:439–441. 1996. View Article : Google Scholar : PubMed/NCBI
30	Dong Z and Chen N: Study on the TLC identification method of Wangbuliuxing tablets. China For Med Treat. 30:1282011.
31	Li N, Ma CH, Liu D, et al: Chemical constituents analysis of fried Vaccaria segetafis. Chin J Exptl Tradit Med Form. 19:73–75. 2013.
32	Sun YX, Liang HT, Zhang XT, et al: Structural elucidation and immunological activity of a polysaccharide from the fruiting body of Armillaria mellea. Biores Technol. 100:1860–1863. 2009. View Article : Google Scholar
33	Sun Y, Wang S, Li T, et al: Purification structure and immunobiological activity of a new water-soluble polysaccharide from the mycelium of Polyporus albicans (Imaz) Teng. Biores Technol. 99:900–904. 2008. View Article : Google Scholar
34	Meng H, Chen Y, Qin W, et al: Determination of vaccarin in Vaccariae Semen by HPLC. Zhongguo Zhong Yao Za Zhi. 35:2072–2074. 2010.PubMed/NCBI
35	Xia MX, Feng L and Zhang LF: Isolation, Purification and Identification of Chemical Compound from Semen vaccariae to Inhibit Endothelial Cells. Biotechnology Bulletin. 2:93–97. 2009.
36	Cheong SM, Choi H, Hong BS, Gho YS and Han JK: Dab2 is pivotal for endothelial cell migration by mediating VEGF expression in cancer cells. Exp Cell Res. 318:550–557. 2012. View Article : Google Scholar : PubMed/NCBI
37	Liu HT, Li WM, Xu G, et al: Chitosan oligosaccharides attenuate hydrogen peroxide-induced stress injury in human umbilical vein endothelial cells. Pharmacol Res. 59:167–175. 2009. View Article : Google Scholar : PubMed/NCBI
38	Li B, Qiu T, Zhang P, Wang X, Yin Y and Li S: IKVAV regulates ERK1/2 and Akt signalling pathways in BMMSC population growth and proliferation. Cell Prolif. 47:133–145. 2014. View Article : Google Scholar : PubMed/NCBI
39	Liang KW, Lee WJ, Lee WL, Chen YT, Ting CT and Sheu WH: Diabetes exacerbates angiographic coronary lesion progression in subjects with metabolic syndrome independent of CRP levels. Clin Chim Acta. 388:41–45. 2008. View Article : Google Scholar
40	Dikalov SI, Dikalova AE, Bikineyeva AT, Schmidt HH, Harrison DG and Griendling KK: Distinct roles of Nox1 and Nox4 in basal and angiotensin II-stimulated superoxide and hydrogen peroxide production. Free Radical Biol Med. 45:1340–1351. 2008. View Article : Google Scholar
41	Li JK, Ge R, Tang L and Li QS: Protective effects of farrerol against hydrogen-peroxide-induced apoptosis in human endothelium-derived EA.hy926 cells. Can J Physiol Pharmacol. 91:733–740. 2013. View Article : Google Scholar : PubMed/NCBI
42	Noseda M, McLean G, Niessen K, et al: Notch activation results in phenotypic and functional changes consistent with endothelial-to-mesenchymal transformation. Circ Res. 94:910–917. 2004. View Article : Google Scholar : PubMed/NCBI
43	Williams CK, Li JL, Murga M, Harris AL and Tosato G: Up-regulation of the Notch ligand Delta-like 4 inhibits VEGF-induced endothelial cell function. Blood. 107:931–939. 2006. View Article : Google Scholar
44	MacKenzie F, Duriez P, Wong F, Noseda M and Karsan A: Notch4 inhibits endothelial apoptosis via RBP-Jkappa-dependent and -independent pathways. J Biol Chem. 279:11657–11663. 2004. View Article : Google Scholar : PubMed/NCBI
45	Ding X, Zhu F, Li T, Zhou Q, Hou FF and Nie J: Numb Protects Renal Proximal Tubular Cells from Puromycin Aminonucleoside-Induced Apoptosis through Inhibiting Notch Signaling Pathway. Int J Biol Sci. 7:269–278. 2011. View Article : Google Scholar : PubMed/NCBI
46	Stampfer MJ, Hennekens CH, Manson JE, Colditz GA, Rosner B and Willett WC: Vitamin E consumption and the risk of coronary disease in women. N Engl J Med. 328:1444–1449. 1993. View Article : Google Scholar : PubMed/NCBI
47	Rimm EB, Stampfer MJ, Ascherio A, Giovannucci E, Colditz GA and Willett WC: Vitamin E consumption and the risk of coronary heart disease in men. N Engl J Med. 328:1450–1456. 1993. View Article : Google Scholar : PubMed/NCBI
48	Chen SD, Yin JH, Hwang CS, Tang CM and Yang DI: Anti-apoptotic and anti-oxidative mechanisms of minocycline against sphingomyelinase/ceramide neurotoxicity: implication in Alzheimer’s disease and cerebral ischemia. Free Radic Res. 46:940–950. 2012. View Article : Google Scholar : PubMed/NCBI
49	Luo P, Chen T, Zhao Y, et al: Protective effect of Homer 1a against hydrogen peroxide-induced oxidative stress in PC12 cells. Free Radic Res. 46:766–776. 2012. View Article : Google Scholar : PubMed/NCBI
50	Fan G, Ma X, Wong PY, Rodrigues CM and Steer CJ: p53 dephosphorylation and p21 (Cip1/Waf1) translocation correlate with caspase-3 activation in TGF-beta1-induced apoptosis of HuH-7 cells. Apoptosis. 9:211–221. 2004. View Article : Google Scholar : PubMed/NCBI
51	Duan W, Yang Y and Yi W: New Role of JAK2/STAT3 signaling in endothelial cell oxidative stress injury and protective effect of melatonin. PLoS One. 8:1–13. 2013. View Article : Google Scholar