Puerarin protects human umbilical vein endothelial cells against high glucose-induced apoptosis by upregulating heme oxygenase-1 and inhibiting calpain activation
Ying-Ying Chen
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Department of Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorJie Chen
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorXin-Mei Zhou
Department of Physiology, Jiaxing University School of Medicine, Jiaxing 314001, China
Search for more papers by this authorXiang-Hong Meng
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorJian-Ping Jiang
Department of Surgery, Zhejiang Medical College, Hangzhou 310053, China
Search for more papers by this authorCorresponding Author
Yue-Liang Shen
Department of Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310058, China
Correspondence and reprints: [email protected]Search for more papers by this authorYing-Ying Chen
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Department of Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorJie Chen
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorXin-Mei Zhou
Department of Physiology, Jiaxing University School of Medicine, Jiaxing 314001, China
Search for more papers by this authorXiang-Hong Meng
National Education Base for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China
Search for more papers by this authorJian-Ping Jiang
Department of Surgery, Zhejiang Medical College, Hangzhou 310053, China
Search for more papers by this authorCorresponding Author
Yue-Liang Shen
Department of Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310058, China
Correspondence and reprints: [email protected]Search for more papers by this authorAbstract
The aim of this study was to investigate whether puerarin protects against high glucose (HG)-induced apoptosis by suppressing calpain activation in human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to normal glucose (NG) (5.5 mm) or HG (33 mm) for 48 h; then, apoptosis and caspase-3 activity were determined. The expression of heme oxygenase-1 (HO-1) mRNA was evaluated by RT-PCR analysis. The activation of calpain and HO activity were also assessed. Compared with the NG group, exposure of HUVECs to HG for 48 h resulted in significant increases in calpain and caspase-3 activity as well as apoptosis, which were prevented by co-incubation with puerarin (1–100 μm) in a concentration-dependent manner. HO-1 mRNA expression and HO activity were decreased in HUVECs treated with HG for 48 h. Compared with the group exposed to HG alone, co-incubation of HUVECs with puerarin and HG induced increases in HO-1 mRNA expression and HO activity. The HO-1 inhibitor protoporphyrin IX zinc (II) abolished the inhibitory effect of puerarin on HG-induced calpain and caspase-3 activation, as well as apoptosis. The data show that puerarin protects against HG-induced endothelial cell apoptosis by a mechanism involving upregulation of HO-1 expression and inhibition of calpain activity.
References
- 1 Boyle P.J. Diabetes mellitus and macrovascular disease: mechanisms and mediators. Am. J. Med. (2007) 120 S12–S17.
- 2 Thomas J.E., Foody J.M. The pathophysiology of cardiovascular disease in diabetes mellitus and the future of therapy. J. Cardiometab. Syndr. (2007) 2 108–113.
- 3 Hartge M.M., Unger T., Kintscher U. The endothelium and vascular inflammation in diabetes. Diab. Vasc. Dis. Res. (2007) 4 84–88.
- 4 Hermans M.P. Diabetes and the endothelium. Acta Clin. Belg. (2007) 62 97–101.
- 5 Wadham C., Parker A., Wang L., Xia P. High glucose attenuates protein S-nitrosylation in endothelial cells: role of oxidative stress. Diabetes (2007) 56 2715–2721.
- 6 Jiang B., Liu J.H., Bao Y.M., An L.J. Hydrogen peroxide-induced apoptosis in pc12 cells and the protective effect of puerarin. Cell Biol. Int. (2003) 27 1025–1031.
- 7 Xiong F.L., Sun X.H., Gan L., Yang X.L., Xu H.B. Puerarin protects rat pancreatic islets from damage by hydrogen peroxide. Eur. J. Pharmacol. (2006) 529 1–7.
- 8 Meng X.H., Ni C., Zhu L., Shen Y.L., Wang L.L., Chen Y.Y. Puerarin protects against high glucose-induced acute vascular dysfunction: role of heme oxygenase-1 in rat thoracic aorta. Vascul. Pharmacol. (2009) 50 110–115.
- 9 Baumgartner-Parzer S.M., Wagner L., Pettermann M., Grillari J., Gessl A., Waldhausl W. High-glucose – triggered apoptosis in cultured endothelial cells. Diabetes (1995) 44 1323–1327.
- 10 Sheu M.L., Ho F.M., Yang R.S. et al. High glucose induces human endothelial cell apoptosis through a phosphoinositide 3-kinase-regulated cyclooxygenase-2 pathway. Arterioscler. Thromb. Vasc. Biol. (2005) 25 539–545.
- 11 Iori E., Pagnin E., Gallo A. et al. Heme oxygenase-1 is an important modulator in limiting glucose-induced apoptosis in human umbilical vein endothelial cells. Life Sci. (2008) 82 383–392.
- 12 Du Y., Miller C.M., Kern T.S. Hyperglycemia increases mitochondrial superoxide in retina and retinal cells. Free Radic. Biol. Med. (2003) 35 1491–1499.
- 13 Hao L.N., Ling Y.Q., Luo X.M. et al. Puerarin decreases lens epithelium cell apoptosis induced partly by peroxynitrite in diabetic rats. Sheng Li Xue Bao (2006) 58 584–592.
- 14 Shen J.G., Yao M.F., Chen X.C., Feng Y.F., Ye Y.H., Tong Z.H. Effects of puerarin on receptor for advanced glycation end products in nephridial tissue of streptozotocin-induced diabetic rats. Mol. Biol. Rep. (2009) 36 2229–2233.
- 15 Zhu L.H., Wang L., Wang D. et al. Puerarin attenuates high-glucose- and diabetes-induced vascular smooth muscle cell proliferation by blocking PKCbeta2/Rac1-dependent signaling. Free Radic. Biol. Med. (2010) 48 471–482.
- 16 Hsu F.L., Liu I.M., Kuo D.H., Chen W.C., Su H.C., Cheng J.T. Antihyperglycemic effect of puerarin in streptozotocin-induced diabetic rats. J. Nat. Prod. (2003) 66 788–792.
- 17 Goll D.E., Thompson V.F., Li H., Wei W., Cong J. The calpain system. Physiol. Rev. (2003) 83 731–801.
- 18 Zatz M., Starling A. Calpains and disease. N. Engl. J. Med. (2005) 352 2413–2423.
- 19 Tie L., Xu Y., Lin Y.H. et al. Down-regulation of brain-pancreas relative protein in diabetic rats and by high glucose in PC12 cells: prevention by calpain inhibitors. J. Pharmacol. Sci. (2008) 106 28–37.
- 20 Li Y., Li Y., Feng Q., Arnold J.M., Peng T. Calpain activation contributes to hyperglycemia-induced apoptosis in cardiomyocytes. Cardiovasc. Res. (2009) 84 100–110.
- 21 Harwood S.M., Allen D.A., Raftery M.J., Yaqoob M.M. High glucose initiates calpain-induced necrosis before apoptosis in LLC-PK1 cells. Kidney Int. (2007) 71 655–663.
- 22 Stalker T.J., Gong Y., Scalia R. The calcium-dependent protease calpain causes endothelial dysfunction in type 2 diabetes. Diabetes (2005) 54 1132–1140.
- 23 Suzuki K., Hata S., Kawabata Y., Sorimachi H. Structure, activation, and biology of calpain. Diabetes (2004) 53 S12–S18.
- 24 Tamareille S., Mignen O., Capiod T., Rucker-Martin C., Feuvray D. High glucose-induced apoptosis through store-operated calcium entry and calcineurin in human umbilical vein endothelial cells. Cell Calcium (2006) 39 47–55.
- 25 Wang S., Peng Q., Zhang J., Liu L. Na+/H+ exchanger is required for hyperglycaemia-induced endothelial dysfunction via calcium-dependent calpain. Cardiovasc. Res. (2008) 80 255–262.
- 26 Ray S.K., Fidan M., Nowak M.W., Wilford G.G., Hogan E.L., Banik N.L. Oxidative stress and Ca2+ influx upregulate calpain and induce apoptosis in PC12 cells. Brain Res. (2000) 852 326–334.
- 27 Gao G., Dou Q.P. N-terminal cleavage of bax by calpain generates a potent proapoptotic 18-kDa fragment that promotes bcl-2-independent cytochrome C release and apoptotic cell death. J. Cell. Biochem. (2000) 80 53–72.
- 28 Raynaud F., Marcilhac A. Implication of calpain in neuronal apoptosis. A possible regulation of Alzheimer’s disease. Febs J. (2006) 273 3437–3443.
- 29 Kim K.A., Lee Y.A., Shin M.H. Calpain-dependent calpastatin cleavage regulates caspase-3 activation during apoptosis of Jurkat T cells induced by Entamoeba histolytica. Int. J. Parasitol. (2007) 37 1209–1219.
- 30 Tan Y., Dourdin N., Wu C., De Veyra T., Elce J.S., Greer P.A. Ubiquitous calpains promote caspase-12 and JNK activation during endoplasmic reticulum stress-induced apoptosis. J. Biol. Chem. (2006) 281 16016–16024.
- 31 Liu Z., Wang S., Zhou H., Yang Y., Zhang M. Na+/H+ exchanger mediates TNF-alpha-induced hepatocyte apoptosis via the calpain-dependent degradation of Bcl-xL. J. Gastroenterol. Hepatol. (2009) 24 879–885.
- 32 Kirkby K.A., Adin C.A. Products of heme oxygenase and their potential therapeutic applications. Am. J. Physiol. Renal. Physiol. (2006) 290 F563–F571.
- 33 Yao P., Hao L., Nussler N. et al. The protective role of HO-1 and its generated products (CO, Bilirubin and Fe) in ethanol-induced human hepatocyte damage. Am. J. Physiol. Gastrointest. Liver Physiol. (2009) 296 G1318–G1323.
- 34 Wu C.C., Lu K.C., Chen J.S. et al. HO-1 induction ameliorates experimental murine membranous nephropathy: anti-oxidative, anti-apoptotic and immunomodulatory effects. Nephrol. Dial. Transplant. (2008) 23 3082–3090.
- 35 De Backer O., Elinck E., Blanckaert B., Leybaert L., Motterlini R., Lefebvre R.A. Water-soluble CO-releasing molecules reduce the development of postoperative ileus via modulation of MAPK/HO-1 signalling and reduction of oxidative stress. Gut (2009) 58 347–356.
- 36 Di Filippo C., Marfella R., Cuzzocrea S. et al. Hyperglycemia in streptozotocin-induced diabetic rat increases infarct size associated with low levels of myocardial HO-1 during ischemia/reperfusion. Diabetes (2005) 54 803–810.
- 37 Abraham N.G., Kushida T., McClung J. et al. Heme oxygenase-1 attenuates glucose-mediated cell growth arrest and apoptosis in human microvessel endothelial cells. Circ. Res. (2003) 93 507–514.
- 38 Quan S., Kaminski P.M., Yang L. et al. Heme oxygenase-1 prevents superoxide anion-associated endothelial cell sloughing in diabetic rats. Biochem. Biophys. Res. Commun. (2004) 315 509–516.
- 39 Drummond G.S., Kappas A. Prevention of neonatal hyperbilirubinemia by tin protoporphyrin IX, a potent competitive inhibitor of heme oxidation. Proc. Natl. Acad. Sci. U S A (1981) 78 6466–6470.
- 40 Kato H., Amersi F., Buelow R. et al. Heme oxygenase-1 overexpression protects rat livers from ischemia/reperfusion injury with extended cold preservation. Am. J. Transplant. (2001) 1 121–128.
- 41 Hwang Y.P., Jeong H.G. Mechanism of phytoestrogen puerarin-mediated cytoprotection following oxidative injury: estrogen receptor-dependent up-regulation of PI3K/Akt and HO-1. Toxicol. Appl. Pharmacol. (2008) 233 371–381.
- 42 Lin C.C., Chiang L.L., Lin C.H. et al. Transforming growth factor-beta1 stimulates heme oxygenase-1 expression via the PI3K/Akt and NF-kappaB pathways in human lung epithelial cells. Eur. J. Pharmacol. (2007) 560 101–109.
- 43 Zhang Y., Guan L., Wang X., Wen T., Xing J., Zhao J. Protection of chlorophyllin against oxidative damage by inducing HO-1 and NQO1 expression mediated by PI3K/Akt and Nrf2. Free Radic. Res. (2008) 42 362–371.
- 44 Zhang S.Y., Chen G., Wei P.F. et al. The effect of puerarin on serum nitric oxide concentration and myocardial eNOS expression in rats with myocardial infarction. J. Asian. Nat. Prod. Res. (2008) 10 373–381.
- 45 Zhang Y., Zeng X., Zhang L., Zheng X. Stimulatory effect of puerarin on bone formation through activation of PI3K/Akt pathway in rat calvaria osteoblasts. Planta Med. (2007) 73 341–347.
- 46 Kim K.M., Jung D.H., Jang D.S. et al. Puerarin suppresses AGEs-induced inflammation in mouse mesangial cells: a possible pathway through the induction of heme oxygenase-1 expression. Toxicol. Appl. Pharmacol. (2010) 244 106–113.
- 47 De Vriese A.S., Verbeuren T.J., Van de Voorde J., Lameire N.H., Vanhoutte P.M. Endothelial dysfunction in diabetes. Br. J. Pharmacol. (2000) 130 963–974.
- 48 Davies P.F. Flow-mediated endothelial mechanotransduction. Physiol. Rev. (1995) 75 519–560.
- 49 Belmadani S., Palen D.I., Gonzalez-Villalobos R.A., Boulares H.A., Matrougui K. Elevated epidermal growth factor receptor phosphorylation induces resistance artery dysfunction in diabetic db/db mice. Diabetes (2008) 57 1629–1637.
- 50 Su J., Lucchesi P.A., Gonzalez-Villalobos R.A. et al. Role of advanced glycation end products with oxidative stress in resistance artery dysfunction in type 2 diabetic mice. Arterioscler. Thromb. Vasc. Biol. (2008) 28 1432–1438.
- 51 Brower J.B., Targovnik J.H., Caplan M.R., Massia S.P. High glucose-mediated loss of cell surface heparan sulfate proteoglycan impairs the endothelial shear stress response. Cytoskeleton (Hoboken) (2010) 67 135–141.
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