Effect of propofol postconditioning on necroptosis during hypoxia-reoxygenation injury in diabetic cardiomyocytes / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the effect of propofol postconditioning on necroptosis during hy-poxia-reoxygenation (H∕R) injury in diabetic cardiomyocytes. Methods Normally cultured H9C2 cardio-myocytes were divided into 5 groups (n= 19 each) using a random number table control group (group C), high glucose group (group HG), H∕R group, propofol postconditioning (group P) and solvent dimethyl sulfoxide (DMSO) control group (group DMSO). H9C2 cells were incubated for 48 h in DMEM culture medium containing 5. 5 and 25 mmol∕L glucose in group C and group HG, respectively. In group H∕R, H9C2 cells were incubated for 48 h in DMEM culture medium containing 25 mmol∕L glucose and then un-derwent H∕R. H9C2 cells were incubated for 48 h in DMEM culture medium containing 25 mmol∕L glucose and then underwent H∕R, and propofol at the final concentration of 50 μmol∕L was added at the onset of reoxygenation in group P. In group DMSO, H9C2 cells were incubated for 48 h in DMEM culture medium containing 25 mmol∕L glucose and then underwent H∕R, and DMSO at the final concentration of 150μmol∕L was added at the onset of reoxygenation. The model of cardiomyocyte H∕R injury was established by subjecting cardiomyocytes to 6 h of hypoxia followed by 12 h of reoxygenation. At 12 h of reoxygenation, the cell viability was measured by CCK8 assay, the product of lactate dehydrogenase (LDH) in culture medium was measured, the level of reactive oxygen species (ROS) was determined by flow cytometry, cardiomyo-cyte apoptosis was detected by TUNEL, and the expression of receptor-interacting protein 1 ( RIP1), RIP3, Bax, Bcl-2, activated caspase-3 and caspase-3 was determined by Western blot. The apoptotic rate and ratio of activated caspase-3∕caspase-3 were calculated. Results Compared with group C, the cell via-bility was significantly decreased, the product of LDH was increased, the level of ROS and apoptotic rate were increased, the expression of RIP1, RIP3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, and the ratio of activated caspase-3∕caspase-3 was increased in group HG ( P < 0. 05). Compared with group HG, the cell viability was significantly decreased, the product of LDH was increased, the level of ROS and apoptotic rate were increased, the expression of RIP1, RIP3 and Bax was up-regula-ted, the expression of Bcl-2 was down-regulated, and the ratio of activated caspase-3∕caspase-3 was in-creased in group H∕R (P<0. 05). Compared with group H∕R, the cell viability was significantly increased, the product of LDH was decreased, the level of ROS and apoptotic rate were decreased, the expression of RIP1, RIP3 and Bax was down-regulated, the expression of Bcl-2 was up-regulated, and the ratio of acti-vated caspase-3∕caspase-3 was decreased in group P (P<0. 05), and no significant change was found in the indexes mentioned above in group DMSO (P>0. 05). Conclusion The mechanism by which propofol post-conditioning ameliorates H∕R injury in diabetic cardiomyocytes may be related to inhibiting necroptosis.