To investigate the primary neurons autophagy and apoptosis after Oxygen Glucose Deprivation/Re-oxygenation (OGD/R)

and to explore the effect of mild hypothermia on autophagy and apoptosis

following methods were used: Primary neurons culture and OGD/R model was established; the neurons were divided into the normal temperature group (37 ℃) and the mild hypothermia group (MH

34 ℃); The cells viability were measured by CCK8; cell apoptosis were measured by TUNEL; The protein expressions of Sirt1

Foxo1

p53

Rab7 and autophagy related genes such as Beclin1

LC3 were detected by western blot at each time point; The neurons autophagy flows were detected through transfection of adenovirus mRFP-GFP-LC3. The primary neuron cultures were successfully developed

and the OGD/R models were established; Westernblot showed that the expressions of Sirt1

P-Foxo1

Rab7

Beclin1 and LC3b/LC3a were gradually reduced

especially at 12 h after OGD/R

P<0.05; However

the expression of p53 was increased

P<0.05; In MH group

the expressions of Sirt1

P-Foxo1

Rab7

Beclin1

LC3b/LC3a were obviously higher than those in NT group

P<0.05; And the expression of p53 was obviously lower than that in NT group

P<0.05; in R6 h+MH group， the rate of neuron cells apoptosis were 20%±6.7%

lower than 56.8%±7.6% in R6 h+NT group

P<0.05; mRFPGFPLC3 adenovirus was transfected into primary neurons

the autophagy flow was detected by fluorescence microscopy. Compared with control group

the autolysosomes were reduced

but autophagosomes were increased after OGD/R

P<0.05; however

compared with NT group

the autophagosomes were reduced and the autolysosomes were increased in MH group

P<0.05. In conclusion

mild hypothermia therapy could increase the expression of Sirt1

Foxo1

beclin1 and LC3b/LC3a

but decrease the expression of p53

so as to promote autophagy and reduce apoptosis.