The Study On The Expression Of Apoptosis In The Mouse Myocardium/Brain After Ketamine Administration

Aim: To observe the expression of apoptosis in the mouse myocardium/brain after ketamine administration.Methods: Animal model of chronic ketaimine poisoning was established by injectinon of ketamine to mouse through tail vein. The 90 KM mice were randomly divided to control (n=10) and poisoning (n=80) group. The control and the all experiment groups were divided to 1, 2, 4, 8, 12 weeks time point after the ketamine administration. 4mg/kg, 10mg/kg, 20mg/kg, 30mg/kg of ketamine were administrated at each time point. The clinical manifestations of ketamine poisoning were observed; Pathology changes of tissue in heart/brain were observed by light microscope. Ultra-structural changes of cells in heart/brain were observed by transmission electron microscopy (TEM), and the quantitative changes of apoptosis were detected with terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The apoptosis cells are labelled by green fluorescence, and the apoptosis number is counted under fluorescence microscopy. To explore the mechanism underlying apoptosis induce by chronic ketamine administration, the expression of activated Caspase-3 was observed with confocal microscopy.Results: From our observation the clinical manifestations of mice induced by ketamine under different doses were different. The gross appearance of tissue was not special. The nucleu of the poisoning group cells was anachromasis under light microscope. One week after ketamine used, TEM showed cells of myocardium / neuron shrinkage and blebbing, chromatin pyknosis, osmotic swelling of cell membrane and organelles. The positive cell of TUNEL was obviously increased 1 week after ketamine injection (p<0.05). And the peak was in the 1~2 weeks. Immunofluorescence staining showed that Caspase-3 was activated 1 week after ketamine used. And the peak was in the 1weeks.Conclusions: Ketamine could induce myocardium / brain apoptosis in chronic ketamine administration of mice. This apoptosis was dependent on the activation of Caspase-3.