ABSTRACT
Objective To investigate the protective effects of emulsified isoflurane(8% v/v) on myocardial cells after hypoxia/reoxygenation(H/R) in cultured myocardial cells from neonatal rats.Methods Myocardial hypoxia/reoxygenation model was established by culturing myocardial cells primarily isolated from neonatal rats.The myocardial cells were divided into 4 groups,the normal group,model group,fat milk treatment group,and mulsified isoflurane treatment group(0.28 mmol/L).Cellular morphologic changes and pulsation frequency were observed under inverted microscope.Cell activity was determined by XTT assay.The activities of plasma superoxide dismutase(SOD) was measured by xanthine oxidase method and the contents of serum malonicaldehyde(MDA) by thiobarbituric acid method.The activities of lactate dehydrogenase(LDH) in the culture medium after H/R injury was measured.Western blot analysis was used to detect caspase-3 protein expression in every group.Results Compared with normal group,SOD activity was decreased,and MDA content,LDH activity and the expression of caspase-3 protein were increased in model group(P
ABSTRACT
Objective To investigate the effects of emulsified isoflurane on myocardial apoptosis, and the expressions of Bcl-2 and Bax after hypoxia/reoxygenation in cultured myocardial cells of neonatal rats. Methods Myocardial hypoxia/reoxygenation model was established on cultured primary myocardial cells of neonatal rat. The myocardial cells were divided into four groups: the normal group, model group, fat milk group, and emulsified isoflurane group. The cellular morphologic changes and pulsatile frequency of each group were observed under inverted microscope and the ultrastructure of myocardial cells was observed under transmission electron microscope. The apoptotic rate of myocardial cells was determined by flow cytometry, the expression of bcl-2 mRNA and bax mRNA were detected by RT-PCR and the protein expressions of Bcl-2/Bax were observed by Western blot. Results The apoptotic rate were significantly decreased in both emulsified isoflurane and fat milk groups (P0.05). Conclusion Emulsified isoflurane and one of its components, fat milk,can inhibit myocardial apoptosis induced by hypoxia/reoxygenation in cultured myocardial cells. Emulsified isoflurane exerts its anti-apoptosis effect maybe through up-regulating Bcl-2 expressions and down-regulating Bax expressions.