Abstract
Dysfunction of endothelial cell is considered as a major cause of vascular complications in diabetes. Crocetin has been shown to have strong antioxidant activities. In present study, we tested whether crocetin inhibited high glucose-induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs) and to explore its possible mechanism. Exposure to high glucose (33 mM) for 72h induced a pronounced increase in apoptosis compared with normal glucose (5.5 mM), as evaluated by cell chromatin staining with Hoechst 33,258 and cell death detection ELISA. High glucose attenuated activation of Akt and endothelial nitric oxide synthase (eNOS). Crocetin (0.1 μM, 1.0 μM) prevented high glucose-induced apoptosis, which correlates with the increase of activation of p-Akt, following the up-regulation of eNOS and NO production. Pretreatment with phosphatidylinositol 3′ kinase (Pl3K) inhibitor LY294002 or eNOS inhibitor NG-nitro-arginine methyl ester (LN or L-NAME) inhibited crocetin’ effect on p-Akt or eNOS, respectively. For the first time, results of our study suggest that crocetin inhibits high glucose-induced apoptosis, at least partly, via Pl3K/Akt/eNOS pathway in HUVECs and crocetin may exert a beneficial effect in preventing diabetes-associated cardiovascular complications.
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Ahmed, N. N., Grimes, H. L., and Bellacosa, A., Transduction of interleukin-2 antiapoptotic and proliferative signals via Akt protein kinase. Proc. Natl. Acad. Sci. U.S.A., 94, 3627–3632 (1997).
Basta, G., Schmidt, A. M., and Caterina, R. D., Advanced glycation end products and vascular inflammation: implications for accelerated atherosclerosis in diabetes. Cardiovascular Research, 63, 582–592 (2004).
Baumgartner-Parzer, M., Wagner, L., Pettermann, M., Grillari, J., Gessl, A., and Waldhusl, W., high-glucose-triggered apoptosis in cultured endothelial cells. Diabetes, 44, 1323–1327 (1995).
Bhat, N. R. and Zhang, P., Hydrogen peroxide activation of multiple mitogen-activated protein kinase in an oligodendrocyte cell line: role of extracellular signal-regulated kinase in hydrogen peroxide-induced cell death. J. Neurochem., 72, 112–119 (1999).
Burgering, B. M. and Coffer, P. J., Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature, 376, 599–602 (1995).
Chavakis, D., Dernbach, E., and Hermann, C., Oxidized LDL inhibits vascular endothelial growth factor-induced endothelial cell migration by an inhibitory effect on the Akt/endothelial nitric oxide synthase pathway. Circulation, 103, 2102–2107 (2001).
Datt, S. R., Brunet, A., and Greenberg, M. E., Cellular survival: a play in three Akts. Genes. Dev., 13, 2905–2927 (1999).
Dudek, H., Datta, S. R., Franke, T. F., Birnbaum M. J., Yao, R., Cooper, G. M., Segal, R. A., Kaplan, D. R., and Greenberg, M. E., Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science, 275, 661–665 (1997).
Fulton, D., Gratton, J. P., McCabe, T. J., Fontana, J., Walsh, K., and Franke, T. F., Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature, 39, 597–601 (1999).
Giuglano, D., Ceriello, A., and Paolisso, G., Oxidative stress and diabetic vascular complications. Diabetes Care, 19, 257–267 (1996).
Gong, G. Q., Liu, T. Z., and Li, L. W., Antioxidative activity of crocetin in vitro. J. China Pharm. Univ., 32, 306–309 (2001).
Green, L. C., Wagner, D. A., Glogowski, J., Skipper, P. L., Wishnok, J. S., and Tannenbaum, S. R., Analysis of nitrate, nitrite, a [15N]) nitrate in biological fluids. Anal. Biochem., 126, 131–138 (1982).
He, S. Y., Qian, Z. Y., Tang, F. T., Wen, N., Xu, G. L., and Sheng, L., Effect of crocin on experimental atherosclerosis in quails and its mechanisms. Life Sciences, 77, 907–921 (2005).
Ho, F. M., Liu, S. H., and Liau, C. S., Nitric oxide prevents apoptosis of human endothelial cells from high glucose exposure during early stage. J. Cell Biochem., 75, 258–263 (1999).
Ho, F. M., Liu, S. H., and Liau, C. S., High glucose-induced apoptosis in human vascular endothelial cells is mediated through NF-κB and c-Jun NH2-terminal kinase pathway and prevented by Pl3K/Akt/eNOS pathway. Circulation, 101, 2618–2624 (2000).
Ho, F. M., Lin, W. W., Chen, B. C., Chao, C. M., Yang, C. R., and Lin, L. Y., High glucose-induced apoptosis in human vascular endothelial cells is mediated through NF-κB and c-Jun NH2-terminal kinase pathway and prevented by Pl3K/Akt/eNOS pathway. Cell Signal, 18, 391–399 (2006).
Ido, Y., Carling, D., and Ruderman, N., Hyperglycemia-induced apoptosis in human umbilical vein endothelial cells: inhibition by the AMP-activated protein kinase activation. Diabetes, 5, 159–167 (2002).
Jaffe, E. A., Nachman, R. L., Bedker, C. G., and Minick, C. R., Culture of human endothelial cells derived from umbilical veins. J. Clin. Invest., 52, 2745–2756 (1973).
Kim, I., Kim, H. G., and So, J. N., Angiopoietin-1 regulates endothelial cell survival through the phosphatidylinositol 3′-Kinase/Akt signal transduction pathway. Circ. Res., 86, 24–29 (2000).
Liu, J. and Qian, Z., Effects of crocin on cholestane-3,5,6-triol-induced apoptosis and related gene expression of cultured endothelial cells. J. China Pharm. Univ., 36, 254–259 (2005).
Lorenzi, M., Nordberg, J. A., and Toledo, S., High glucose prolongs cell-cycle traversal of cultured human endothelial cells. Diabetes, 36, 1261–1267 (1991).
Mozaffarian, N., Casadevall, A., and Berman, J. W., Inhibition of human endothelial cell chemokine production by the opportunistic fungal pathogen Cryptococcus neoformans. J. Immunol., 165, 1541–1547 (2000).
Pascal, M. M., Knott, R. M., and Forrester, J. V., Glucose mediated regulation of transforming growth factor beta in human retinal endothelial cells. Biochem. Soc. Trans., 24, 228s (1996).
Robertson, R. P., Harmon, J., Tran, P. O., Tanaka, Y., and Takahashi, H., Glucose toxicity in b-cells: type 2 diabetes, good radicals gone bad, and the glutathione connection. Diabetes, 52, 581–587 (2003).
Scheid, M. P. and Woodgett, J. R., Protein kinases: six degrees of separation? Curr. Biol., 10, R191–R194 (2000).
Sheu, M. L., Ho, F. M., Yang, R. S., Chao, K. F., Lin, W. W., Lin-Shiau, S. Y., and Liu, S. H., Glucose induces human endothelial cell apoptosis through a phosphoinositide 3-kinase-regulated cyclooxygenase-2 pathway. Arterioscler. Thromb. Vasc. Biol., 25, 539–545 (2005).
Sophie, T., Olivier, M., Thierry, C., Catherine, R. M., and Danielle, F., High glucose-induced apoptosis through storeoperated calcium entry and calcineurin in human umbilical vein endothelial cells. Cell Calcium, 39, 47–55 (2006).
Tang, F. T., Qian, Z. Y., and Zheng, S. G., Effects of crocetin on serum nitric oxide content of atherosclerotic quail and its mechanism. Chin. J. Nat. Med., 3, 316–320 (2005).
Tesfamariam, B. and Cohen, R. A., Free radicals mediate endothelial cell dysfunction caused by elevated glucose. Am. J. Physiol., 263, 321–326 (1992).
Wang, H. and Joseph, J. A., Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radical Biology and Medicine, 27, 612–616 (1999).
Wang, T. T. and Phang, J. M., Effects of estrogen on apoptotic pathways in human breast cancer cell line MCF-7. Cancer Res., 55, 2487–2489 (1955).
Wang, X., Martindale, J. L., Liu, Y., and Holbrook, N. J., The cellular response to oxidative stress: inuences of mitogenactivated protein kinase signaling pathways on cell survival. Biochem. J., 333, 291–300 (1998).
Xiang, M., Yang, M., Zhou, C. H., Liu, J., Li, W. N., and Qian, Z. Y., Crocetin prevents AGEs-induced vascular endothelial cell apoptosis. Pharmacological Research, 54, 268–274 (2006).
Yokozawa, T., Kim, Y. A., Kim, H. Y., Lee, Y. A., and Nonaka, G., Protective effect of persimmon peel polyphenol against high glucose-induced oxidative stress in LLC-PK1 cells. Food and Chemical Toxicology, 45, 1979–1987 (2007).
Yu, W., Qian, Z., Xu, G., and Shen, C., Effects of crocetin on the myocardial cell damages due to oxidative stress. J. China Pharm. Univ., 34, 452–455 (2003).
Zhang, W., Wang, R., Han, S. F., Bu, L., Wang, S. W., Ma, H., and Jia, G. I., α-Linolenic acid attenuates high glucose-induced apoptosis in cultured human umbilical vein endothelial cells via Pl3K/Akt/eNOS pathway. Nutrition, 23, 762–770 (2007).
Zheng, S., Qian, Z., and Tang, F., Suppression of vascular cell adhesion molecule-1 expression by crocetin contributes to attenuation of atherosclerosis in hypercholesterolemic rabbits. Biochem. Pharmacol., 70, 1192–1199 (2005).
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Meng, L., Cui, L. Inhibitory effects of crocetin on high glucose-induced apoptosis in cultured human umbilical vein endothelial cells and its mechanism. Arch. Pharm. Res. 31, 357–363 (2008). https://doi.org/10.1007/s12272-001-1164-y
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DOI: https://doi.org/10.1007/s12272-001-1164-y