Safflor yellow A protects neonatal rat cardiomyocytes against anoxia/reoxygenation injury in vitro
Abstract
Jia-lin DUAN#, Jing-wen WANG#, Yue GUAN#, Ying YIN#, Guo WEI#, Jia CUI, Dan ZHOU, Yan-rong ZHU, Wei QUAN, Miao-miao XI*, Ai-dong WEN*
Department of Pharmacy, Xijing Hospital, the Fourth Military Medical University, Xi-an 710032, China
Aim: To investigate the effects of safflor yellow A (SYA), a flavonoid extracted from Carthamus tinctorius L, on cultured rat cardiomyocytes exposed to anoxia/reoxygenation (A/R).
Methods: Primary cultured neonatal rat cardiomyocytes were exposed to anoxia for 3 h followed by reoxygenation for 6 h. The cell viability was measured using MTT assay. The releases of lactate dehydrogenase (LDH) and creatine kinase (CK), level of malondialdehyde (MDA), and activities of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were analyzed. Hoechst 33258 staining and changes in Bcl-2/Bax ratio and caspase 3 activity were used to examine A/R-induced apoptosis.
Results: The A/R exposure markedly decreased the viability of cardiomyocytes, suppressed the activities of SOD, GSH, CAT and GSH-Px, and Bcl-2 protein expression. Meanwhile, the A/R exposure markedly increased the release of LDH and CK, and MDA production in the cardiomyocytes, and increased the rate of apoptosis, caspase 3 activity, Bax protein expression. Pretreatment with SYA (40, 60 and 80 nmol/L) concentration-dependently blocked the A/R-induced changes in the cardiomyocytes. Pretreatment of the cardiomyocytes with the antioxidant N-acetylcysteine (NAC, 200 μmol/L) produced protective effects that were comparable to those caused by SYA (80 nmol/L).
Conclusion: SYA protects cultured rat cardiomyocytes against A/R injury, maybe via inhibiting cellular oxidative stress and apoptosis.
Keywords: safflor yellow A; N-acetylcysteine; antioxidant; cardiomyocyte; anoxia/reoxygenation; oxidative stress; apoptosis
This work was financially supported by National Natural Science Foundation of China (No 81173514 and 81001673), the “13115” Technology Innovation Project of Shaanxi Province (No 2010ZDKG-62), Xijing Research Boosting Program (No XJZT10D02) and Excellent Civil Service Training Fund of the Fourth Military Medical University (No 4138C4IDK6). The authors thank colleagues in the New Drug Research and Development Center of Xijing Hospital for their technical assistance.
# These authors contributed equally to this work.
* To whom correspondence should be addressed.
E-mail adwen-2012@hotmail.com (Ai-dong WEN); handsomfish@yahoo.com.cn (Miao-miao XI)
Received 2012-08-08 Accepted 2012-12-06
Keywords:
Department of Pharmacy, Xijing Hospital, the Fourth Military Medical University, Xi-an 710032, China
Aim: To investigate the effects of safflor yellow A (SYA), a flavonoid extracted from Carthamus tinctorius L, on cultured rat cardiomyocytes exposed to anoxia/reoxygenation (A/R).
Methods: Primary cultured neonatal rat cardiomyocytes were exposed to anoxia for 3 h followed by reoxygenation for 6 h. The cell viability was measured using MTT assay. The releases of lactate dehydrogenase (LDH) and creatine kinase (CK), level of malondialdehyde (MDA), and activities of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were analyzed. Hoechst 33258 staining and changes in Bcl-2/Bax ratio and caspase 3 activity were used to examine A/R-induced apoptosis.
Results: The A/R exposure markedly decreased the viability of cardiomyocytes, suppressed the activities of SOD, GSH, CAT and GSH-Px, and Bcl-2 protein expression. Meanwhile, the A/R exposure markedly increased the release of LDH and CK, and MDA production in the cardiomyocytes, and increased the rate of apoptosis, caspase 3 activity, Bax protein expression. Pretreatment with SYA (40, 60 and 80 nmol/L) concentration-dependently blocked the A/R-induced changes in the cardiomyocytes. Pretreatment of the cardiomyocytes with the antioxidant N-acetylcysteine (NAC, 200 μmol/L) produced protective effects that were comparable to those caused by SYA (80 nmol/L).
Conclusion: SYA protects cultured rat cardiomyocytes against A/R injury, maybe via inhibiting cellular oxidative stress and apoptosis.
Keywords: safflor yellow A; N-acetylcysteine; antioxidant; cardiomyocyte; anoxia/reoxygenation; oxidative stress; apoptosis
This work was financially supported by National Natural Science Foundation of China (No 81173514 and 81001673), the “13115” Technology Innovation Project of Shaanxi Province (No 2010ZDKG-62), Xijing Research Boosting Program (No XJZT10D02) and Excellent Civil Service Training Fund of the Fourth Military Medical University (No 4138C4IDK6). The authors thank colleagues in the New Drug Research and Development Center of Xijing Hospital for their technical assistance.
# These authors contributed equally to this work.
* To whom correspondence should be addressed.
E-mail adwen-2012@hotmail.com (Ai-dong WEN); handsomfish@yahoo.com.cn (Miao-miao XI)
Received 2012-08-08 Accepted 2012-12-06