Title: PUMA-targeting siRNA Protects Rat Cardiomyocyte from Hypoxia/Reoxygenation-induced Apoptosis
Abstract: To study the effect of p53 up-regulated modulator of apoptosis(PUMA) in hypoxia/ reoxygenation(H/R) mediated rat cardiomyocyte injury,siRNA targeted PUMA was designed and transfected into primary cultured rat cardiomyocytes in parallel with a scramble siRNA.Optimized dose of PUMA-specific siRNA was 50 nmol/L as determined by RT-PCR and Western blotting.The MTT assay and lactate dehydrogenase(LDH) activity measurement showed that the cell survival was enhanced and LDH release was reduced after the transfection of PUMA siRNA(P0.01,compared with the control).PUMA siRNA could also down-regulate caspase-3 activity and decrease apoptosis rate(P0.01) from the results of annexin V apoptosis assay and PI-stained with flow cytometry.Down-regulation of Bax and up-regulation of Bcl-2 were detected in RT-PCR(P0.05).These results indicated that PUMA siRNA could protect cardiomyocytes from apoptosis and could be considered for the therapy for cardiac ischemia/reperfusion injury.
Publication Year: 2011
Publication Date: 2011-01-01
Language: en
Type: article
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