Protection of Luteolin-7-O-Glucoside Against Doxorubicin-Induced Injury Through PTEN/Akt and ERK Pathway in H9c2 Cells
Luteolin-7-O-glucoside (LUTG) was isolated from the plants of Dracocephalum tanguticum Maxim. Previous research has showed that LUTG pretreatment had a significant protective effect against doxorubicin (DOX)-induced cardiotoxicity by reducing intracellular calcium overload and leakage of creatine kinase and lactate dehydrogenase. But the underlying mechanisms have not been completely elucidated. In the present study, we investigated the effects of LUTG on H9c2 cell morphology, viability, apoptosis, reactive oxygen species generation, and the mitochondrial transmembrane potentials. The expression of p-PTEN, p-Akt, p-ERK, p-mTOR, and p-GSK-3β were detected by Western blotting. Compared with DOX alone treatment group, the morphological injury and apoptosis of the cells in groups treated by DOX plus LUTG were alleviated, cell viability was increased, ROS generation was lowered remarkably, and mitochondrial depolarization was mitigated. In DOX group, the expression of p-PTEN was lower than normal group and the expression of p-Akt and p-ERK was higher than normal group. In the groups treated with LUTG (20 μM), the expression of p-PTEN was upregulated and the expression of p-Akt, p-ERK, p-mTOR, and p-GSK-3β was downregulated. These results indicated that the protective effects of LUTG against DOX-induced cardiotoxicity may be related to anti-apoptosis through PTEN/Akt and ERK pathway.
KeywordsLuteolin-7-O-glucoside Doxorubicin PTEN Akt ERK
This work was supported by Grants from the Natural Science Foundation of Shandong Province (No. ZR2013HM084) and Independent Innovation Foundation of Shandong University (No. 2012ZD043) of P.R.China.
Conflict of interest
- 2.Santacruz, L., Darrabie, M. D., Mantilla, J. G., Mishra, R., Feger, B. J., & Jacobs, D. O. (2014). Creatine supplementation reduces doxorubicin-induced cardiomyocellular injury. Cardiovascular Toxicology. doi: 10.1007/s12012-014-9283-x.
- 5.Wu, Z. Y., & Li, X. W. (2005). Flora of China (vol. 65). Beijing: Science Press.Google Scholar
- 10.Shioda, N., Han, F., Moriguchi, S., & Fukunaga, K. (2007). Constitutively active calcineurin mediates delayed neuronal death through Fas-ligand expression via activation of NFAT and FKHR transcriptional activities in mouse brain ischemia. Journal of Neurochemistry, 102, 1506–1517.CrossRefPubMedGoogle Scholar
- 11.Shioda, N., Ishigami, T., Han, F., Moriguchi, S., Shibuya, M., Iwabuchi, Y., & Fukunaga, K. (2007). Activation of phosphatidylinositol 3-kinase/protein kinase B pathway by a vanadyl compound mediates its neuroprotective effect in mouse brain ischemia. Neuroscience, 148, 221–229.CrossRefPubMedGoogle Scholar
- 13.Siddall, H. K., Warrell, C. E., Yellon, D. M., & Mocanu, M. M. (2008). Ischemia-reperfusion injury and cardioprotection: investigating PTEN, the phosphatase that negatively regulates PI3K, using a congenital model of PTEN haploinsufficiency. Basic Research in Cardiology, 103, 560–568.CrossRefPubMedGoogle Scholar
- 19.Kim, S. Y., Kim, S. J., Kim, B. J., Rah, S. Y., Chung, S. M., Im, M. J., & Kim, U. H. (2006). Doxorubicin-induced reactive oxygen species generation and intracellular Ca2+ increase are reciprocally modulated in rat cardiomyocytes. Experimental & Molecular Medicine, 38, 535–545.CrossRefGoogle Scholar
- 28.Théberge, J. F., Mehdi, M. Z., Pandey, S. K., & Srivastava, A. K. (2003). Prolongation of insulin-induced activation of mitogen-activated protein kinases ERK 1/2 and phosphatidylinositol 3-kinase by vanadyl sulfate, a protein tyrosine phosphatase inhibitor. Archives of Biochemistry and Biophysics, 420, 9–17.CrossRefPubMedGoogle Scholar
- 30.Negoro, S., Oh, H., Tone, E., Kunisada, K., Fujio, Y., Walsh, K., et al. (2001). Glycoprotein 130 regulates cardiac myocyte survival in doxorubicin induced apoptosis through phosphatidylinositol 3-kinase/Akt phosphorylation and Bcl-xL/caspase-3 interaction. Circulation, 103, 555–561.CrossRefPubMedGoogle Scholar
- 33.Ohori, K., Miura, T., Tanno, M., Miki, T., Sato, T., Ishikawa, S., et al. (2008). Ser9 phosphorylation of mitochondrial GSK-3beta is a primary mechanism of cardiomyocyte protection by erythropoietin against oxidant-induced apoptosis. American Journal of Physiology Heart and Circulatory Physiology, 295, H2079–2086.CrossRefPubMedGoogle Scholar
- 34.Angelini, A., Di Ilio, C., Castellani, M. L., Conti, P., & Cuccurullo, F. (2010). Modulation of multidrug resistance p-glycoprotein activity by flavonoids and honokiol in human doxorubicin-resistant sarcoma cells (MES-SA/DX-5): implications for natural sedatives as chemosensitizing agents in cancer therapy. Journal of Biological Regulators and Homeostatic Agents, 24, 197–205.PubMedGoogle Scholar