ABSTRACT
Objective To evaluate the effects of apolipoprotein-J(ApoJ)on hypoxia/reoxygenation(H/R)induced neonatal rat ventricular cells(NRVCs)injury and its related signaling pathways.Methods ApoJ high expression was achieved by infection with recombinant adenovirus in NRVCs.The three gas incubator was used to establish H/R model,SOD mimetic Mn(Ⅲ) tetrakis(4-benzoic acid)porphyrin chloride and eIF2α dephosphory inhibitor Salubrinal were performed the pretreatment.The NRVCs were divided into four groups:normal control group,H/R,ApoJ group,ApoJ+ H/R group,Mn(Ⅲ)TBAP+H/R group and Salubrinal +-H/R group.The cell viability was measured by MTT assay;the leakages of LDH,the expression of SOD and caspase-3/7 activity were detected by ELISA.The protein expression levels of ApoJ,Nox2/gp91phox,caspase-12,CHOP,and the phosphorylation level of eukaryotic initiation factor 2α(eIF2α)were determined by Western blot.Results ApoJ protein in myocardial cells was highly expressed after infection by recombinant adenoviru.Compared with the control group,the cell viability and the activity of SOD were significantly decreased,the leakages of LDH and the activity of caspase-3/7 were increased in the H/R group,the pro tein expression level of Nox2/gp91phox,caspase-12 and CHOP and the phosphorylation level of eIF2α were increased.Compared with the H/R group,the leakages of LDH and the activity of caspase-3/7 in the ApoJ overexpression group,Mn(Ⅲ)TBAP group and Salubirnal group were significantly decreased.ApoJ overexpression significantly increased the cell viability and the activity of SOD.Moreover,the protein expression level of Nox2/gp91phox,caspase-12 and CHOP were significantly decreased,while the phosphorylation level of eIF2α was markedly increased.Conclusion ApoJ alleviates H/R induced myocardial cellular injury by antioxidative stress and endoplasmic reticulum stress.