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Objective: The study of ketamine on glutamate-induced rat astrocytes autophagy. Methods: newborn rat cerebral cortex, primary mixed culture isolated, purified astrocytes, experimental design of three groups: a control group (C group, joined the D-Hank's solution); glutamic acid group (G group , adding glutamic acid to a final concentration of 125μmol / L); glutamate and ketamine group (GK group, first by adding glutamic acid to a final concentration of 125μmol / L, 30min ketamine were added to a final concentration of 1 mmol / L,), the latter two The groups were designed six time points. Morphological changes were observed by phase contrast microscopy, immunofluorescence staining techniques to identify cell types and detect LC3 protein expression by Western blot detection beclin1 bcl2 LC3 protein expression changes. Results: Immunofluorescence staining test results show, LC3 expression located in the cytoplasm; LC3 Ⅱ expression was scattered distribution group C visibility obvious expression of LC3 Ⅱ in rare the LC3 Ⅱ, G group, GK group than in the group G expression weakened. Western blot results of quantitative analysis, The G cells beclin1, beclin1/bcl2, LC3 Ⅱ / LC3 Ⅰ protein expression is increased compared with C group, bcl2 protein expression compared with group C decreased GK cells beclin1, beclin1/bcl2, LC3 Ⅱ / LC3 Ⅰ protein compared with the G group decreased, bcl2 protein than G group rose. Conclusion: 125μmol / L glutamate can promote astrocyte autophagy, 1 mmol / L ketamine inhibits glutamate-induced autophagy astrocytes play a certain brain protective effect. The second part of the purpose: research repeatedly after intraperitoneal injection of low-dose ketamine hippocampus autophagy-related protein LC3, expression of beclin1 and the significance. : Adult SD (Sprague-Dawley) rats were randomly divided into a treatment group and the saline group, medication rats 5mg/kg dose intraperitoneal injection of ketamine, injected once every half hour, a total of five times. Saline group were given normal saline. Treatment group rats were divided into 6 groups according to the time after treatment, respectively 1,3,6,12,24,48 hours after the last administration to take hippocampus spare, Western blot analysis in the rat hippocampus LC3 beclin1 expression, comparing medication group and the saline group protein expression differences. Results: Compared with the saline group, the treatment group rat hippocampal tissue LC3 Ⅱ / LC3 Ⅰ values ??began to increase in 1 hour, 6 hours was strongly expressed in beclinl expression began to increase at 6 hours, 12 hours, 24 hours, 48 ??hours were tested strong expression (P lt; 0.05). Conclusion: repeatedly intraperitoneal injection of low-dose ketamine can promote hippocampal tissue autophagy Autophagy enhances ketamine toxic reaction.
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