Abstract: Objective To observe the protective effect of Hydroxysafflor yellow A (HSYA) on anoxia/reoxygenation (A/R) injury of neonatal primary cardiomyocytes, and its relationship with phosphoinositide 3-kinase / protein kinase B / glycogen synthase kinase 3β (PI3K/Akt/GSK3β) signaling pathway. Methods The neonatal rats, primary cardiomyocytes were isolated from the rats and incubated for 48 hours. The cells were adhered to each other and then divided into five groups: control group (Con group) , anoxia / reoxygenation group (A/R group) , HSYA treatment group（A/R+H group）, PI3K inhibitor （LY294002）treatment group（A/R+L group）and HSYA + LY294002 treatment group (A/R + H +L group). Collect the supernatant fluid of each group to measure LDH.The flow cytometry was used to measure the rate of apoptotic cells. The protein levels of Bcl-2, Bax, Akt,p-Akt (Ser473),GSK3β ,p-GSK3β (Ser9) was evalated by Western blot. Results A/R increased LDH release ,the apoptosis rate (P<0.001), and the expression of pro-apoptotic protein Bax (P <0.001) with the decrease of anti-apoptotic protein Bcl-2, p-Akt（Ser473）, p-GSK3β（Ser9）(P<0.001) compared with the control group. HSYA treatment decreased LDH release ,the apoptosis rate (P<0.001), and the expression of Bax (P<0.001) and increase the expression of Bcl-2, p-Akt（Ser473）, p-GSK3β（Ser9）(P<0.001) . Compared with the A/R+ H group, the expression of Bax was increased (P<0.001),while the expression of Bcl-2, p-Akt（Ser473）, p-GSK3β（Ser9）was decreased (P<0.001) in the A/R + H + L group. Conclusions HSYA protects rats，cardiomyocytes from anoxia / reoxygenation injury by regulating PI3K/ Akt / GSK3β signaling pathway.

Key words: Anoxia / reoxygenation, HSYA, cardiomyocytes, Apoptosis, PI3K / Akt / GSK3βsignaling pathway