ABSTRACT
Objective: To investigate the effect of Trichosanthis Pericarpium aqueous extract(TPAE)in protecting H9c2 cells from hypoxia/reoxygenation (H/R) injury by activating phosphatidylionsitol-3-kinase/protein kinase B/nitric oxide(PI3K/Akt/NO) signaling pathway. Method: The 2.5 mmol·L-1 Na2S2O4 was used to induce the model of H9c2 cardiomyocytes H/R injury in the experiments. The cultured H9c2 cardiomyocytes were randomly divided into normal group, H/R group (model group) and inhibition group (LY294002, 10 μmol·L-1). In the H/R + TPAE group, 50 mg·L-1 TPAE was added to the cultures at 24 h before H/R exposure. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay. The amounts of NO, endothelial nitric oxide synthase (eNOS), and induced nitric oxide synthase (iNOS) were tested by enzyme linked immunosorbent assay (ELISA) kits. Reverse transcription-quantitative real-time polymerase chain reaction (Real-time PCR) was performed to analyze relative mRNA expressions of Akt, eNOS and iNOS. Western blot was used to detect the expressions of Akt, p-Akt (Ser 473), eNOS, and iNOS. Result: Compared with the normal control group,the cell viability significantly decreased in the model control group (PPPPPPPPConclusion: TPAE can protect H9c2 cardiomyocytes from H/R injury by activating PI3K/Akt signaling pathway,which might be related to the up-regulation of the mRNA and protein expressions of eNOS, the down-regulation of the level of iNOS, and the increase of the production of physiological amounts of NO.