Danshensu protects against ischemia/reperfusion injury and inhibits the apoptosis of H9c2 cells by reducing the calcium overload through the p-JNK-NF-κB-TRPC6 pathway

作者：Meng, Ying; Li, Wei-Zhu; Shi, You-Wei; Zhou, Bing-Feng; Ma, Rong; Li, Wei-Ping^*

来源：International Journal of Molecular Medicine, 2016, 37(1): 258-266.

DOI：10.3892/ijmm.2015.2419

• Danshensu
• ischemia/reperfusion
• transient receptor potential cation
• subfamily C
• member 6
• H9c2 cells
• calcium overload

Ischemia-reperfusion (I/R) plays an important role in myocardial injury. In the present study, we aimed to examine the protective effects of Danshensu (DSS) against I/R injury and to elucidate the underlying mechanisms. For this purpose, H9c2 cells were cultured in hypoxic solution in a hypoxic incubator for 2 h, and then cultured in a high oxygen incubator for various periods of time and pre-treated with or without DSS, ammonium pyrrolidine dithiocarbamate (PDTC) or SP600125 [a c-Jun N-terminal kinase (JNK) inhibitor]. Cell apoptosis and cytosolic free Ca2+ ([Ca2+](i)) levels were analyzed by flow cytometry. The protein expression levels of JNK, phosphorylated (p-)JNK, nuclear factor-kappa B (NF-kappa B) and transient receptor potential cation channel, subfamily C, member 6 (TRPC6) were measured by western blot analysis. The mRNA expression levels of JNK were measured by RT-qPCR. The results revealed that TRPC6 protein expression, the cell apoptotic rate and the [Ca2+](i) levels increased in a time-dependent manner in the H9c2 cells following the induction of I/R injury. The apoptotic rate and TRPC6 protein expression decreased when the cells were treated with DSS prior to the induction of I/R injury. The knockdown of JNK expression by siRNA decreased the p-JNK and TRPC6 protein expression levels in the H9c2 cells subjected to I/R injury. The protein expression levels of p-JNK and NF-kappa B in the nucleus increased significantly when the H9c2 cells were subjected to I/R injury, whereas NF-kappa B expression in the cytoplasm decreased in a time-dependent manner. However, p-JNK, NF-kappa B and TRPC6 protein expression, the [Ca2+](i) level and cell apoptosis decreased when the H9c2 cells were pre-treated with DSS or SP600125. Therefore, our data suggest that DSS prevents myocardial I/R injury by inhibiting p-JNK activation and NF-kappa B translocation, which potentially upregulate TRPC6 expression, increase the [Ca2+](i) level, and result in the apoptosis of H9c2 cells.

• 出版日期2016-1
• 单位安徽医科大学