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Objective To explore the general anesthetic EI on the rat fetal neural stem cell proliferation(NSCs) and the role played by JNK in the influence. Methods The cultured rat fetal NSCs were randomly divided into 6 groups (n=8 each):normal group (group N);fat milk group (group F);EI groups(including 8. 12,9. 80,12. 04 mmol/L EI) , 9. 80 mmol/L EI group+20 μmol/L SP600125 group ( group EISP) . After incubated for 12 hours, the cellular effects of EI and cell viability were evaluated by MTT reduction assay. The apoptotic rate of the rat NSCs were determined by flow cytometry, and the expression of protein Caspase-3 was observed by Western blot. Results There was no statistical difference in cell viability, apoptotic rate and protein expression of Caspase-3 in group N and group F. EI group had higher cell apoptotic rate(P<0. 01), protein expression of Caspase-3 (P<0. 05 ) , but lower cell viability than group N ( P<0. 01 ) . And significant differences were found between three do-ses of EI groups (P<0. 05). Compared with EI group, lower cell apoptotic rate (P<0. 01), protein expression of Caspase-3 but higher cell viability were observed in group EISP ( P<0. 05 ) . Conclusion JNK plays an important role in EI-induced cytotoxicity possibly in a dose-dependent manner.
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OBJECTIVE@#To explore the effect of emulsified isoflurane (EI) on apoptosis of anoxia-reoxygenation neonatal rat cardiomyocytes and relevant protein expression.@*METHODS@#Cardiac muscle anoxia-reoxygenation damage model was established with culture in vitro neonatal rat cardiomyocytes. The cardiomyocytes were divided into control group, model group, fat emulsion group and EI group. The cardiomyocytes apoptosis rates and lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) index standardization were detected after relevant treatment. The expression of apoptosis-related proteins Bel-2, Bax and Caspase-3 were detected with Western blot approach.@*RESULTS@#After hypoxia/reoxygenation (H/R) model was treated by EI, the cells apoptosis rate decreased and was dramatically below the fat emulsion group (P<0.05). Cardiomyocytes biochemical index detection presented that, compared with the control group that the LDH activity and MDA content dramatically increased (P<0.05), while the SOD activity notably decreased (P<0.05); compared with the H/R group, the SOD activity of the fat emulsion group and EI group increased (P<0.05); while the LDH activity and MDA content decreased (P<0.05). And the change of the EI group was more remarkable than the fat emulsion group (P<0.05). The Western blot analysis presented that, compared with the control group, the Bcl-2 protein expression of the other groups significantly decreased (P<0.05), the expressions of Bax protein and Caspase-3 protein increased significantly (P<0.05); compared with H/R group, cardiomyocytes Bcl-2 protein expression of EI group increased significantly (P<0.05), the expressions of Bax protein and Caspase-3 protein decreased significantly (P<0.05), and the change of EI group was more remarkable than the fat emulsion group (P<0.05).@*CONCLUSIONS@#EI can inhabit the apoptosis of anoxia-reoxygenation damage model cardiomyocytes, and may be related to the up-regulation of expression of Bcl-2 and down-regulation of expression of Caspase-3 protein.
Subject(s)
Animals , Rats , Apoptosis , Caspase 3 , Metabolism , Cell Hypoxia , Emulsions , Pharmacology , Isoflurane , Pharmacology , Malondialdehyde , Metabolism , Myocytes, Cardiac , Cell Biology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats, Wistar , Superoxide Dismutase , MetabolismABSTRACT
Objective To examine the expression of ? defensin genes and inflammatory cytokines genes in rat airway epithelium after intraperitoneal injection of emulsified isoflurane.Methods Thirty-six SD rats were assigned into six groups(A-F) randomly.The tissue from rat airway epithelium were harvested at 1 h after intraperitoneal injection of normal saline,30% fat milk and 8% emulsified isofluranes of 4 different dosages(0.25,0.5,1.0 and 1.5 times of ED50).Then expressions of ? defensin(BD-1,BD-2),tumor necrosis factor-alpha(TNF-?),interleukin beta(IL-1?),IL-8 mRNA were measured by semiquantitative polymerase chain reaction(PCR) using GAPDH as an internal standard.Results Except BD-1 and IL-8 mRNA,marked increases in expressions of TNF-?,IL-1? and BD-2 genes were observed with a dosage-dependent manner.Conclusion The expressions of BD-2 genes and inflammatory cytokines genes(IL-1?,TNF-?)in rat airway epithelium increase significantly in a dosage-dependent manner after intraperitoneal injection of emulsified isoflurane.
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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
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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.