Melatonin protects H9C2 cells against ischemia-reperfusion injury via protein kinase C epsilon signaling pathway
Zhang Bin
Zhai Meng-en
Li Bu-ying
Liu Zhen-hua
Chen Zheng-xi
Yu Peng-fei
Yu Li-ming
Liang Hong-liang
Duan Wei-xun
Jin Zhen-xiao
Yu Shi-qiang
Abstract：Objective To investigate the underlying mechanisms of the protective effect of melatonin ( Mel) on simulated is-chemia/reperfusion ( SIR) injury in H9C2 cells and the role of PKC epsilon ( PKCε) signaling pathway in this process. Methods The cultured H9C2 cells were pretreated with Mel ( 100μmol/L) for 12 h and then subjected to SIR ( 45 min ischemia, 4 h reperfusion) . Cell viability and apoptotic index were detected by CCK-8 and TUNEL methods, and the intracellular malondialdehyde ( MDA) con-tent and superoxide dismutase ( SOD) activity were assessed by specialized kits. The translocation of protein kinase C epsilon ( PKCε) and the expression levels of Bcl-2, Bax, Caspase-3 and oxidative stress marker gp91phox were detected by Western blotting. PKCεspe-cific inhibitor εV1-2 was used to study the role of PKCεin this process. Results We found that SIR treatment reduced cell viability and increased apoptotic index, inhibited PKCεtranslocation and reduced the Bcl-2/Bax ratio. Mel pretreatment significantly increased cell viability and reduced apoptotic index, down-regulated the MDA content and up-regulated the SOD activity. In addition, Mel pre-treatment significantly increased PKCε translocation and Bcl-2/Bax ratio and decreased the expression levels of Caspase-3 and gp91phox . However, these protective effects of Mel mentioned above were largely abolished byεV1-2 administration. Conclusion Mel pretreatment may protects H9C2 cells against SIR injury via attenuating oxidative stress and apoptosis in a PKCε-dependent manner.
Keywords：MelatoninIschemia reperfusion injuryProtein kinase C epsilonOxidative stressApoptosis
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 119-124 )