1.Resveratrol attenuates hypoxia-reperfusion injury induced rat myocardium microvascular endothelial cell dysfunction through upregulating PI3K/Akt/SVV pathways.
Huirong ZUO ; Dongchu LIAO ; Long LIN ; Rongqing ZHANG ; Xiujuan LI
Chinese Journal of Cardiology 2014;42(8):670-674
OBJECTIVETo detect the role of surviving (SVV) in the protective effect of resveratrol against hypoxia/reperfusion injury (H/RI) of cardiac microvascular endothelial cells (CMECs).
METHODSCMECs isolated from the hearts of adult rats were exposed to hypoxia (94% N₂, 5% CO₂, 1% O₂) for 2 h followed by 4 h reoxygenation (95% O₂, 5% CO₂). The cell proliferation of CMECs was measured by MTT assay and Transwell method was used to detect migration ability of CMEC, PI-AnnexinV double staining and flow cytometry technique were employed to observe the apoptotic rate of CMECs. The SVV protein expression was detected with Western blot method.
RESULTSCompared to control group, the proliferation (0.19 ± 0.03 vs. 0.42 ± 0.07, P < 0.01) and migration ((28 ± 2)/5HPF vs. (50 ± 3)/5 HPF, P < 0.01) abilities were impaired and the apoptosis index ((19.7 ± 0.8)% vs. (5.4 ± 0.3)%, (P < 0.05) of CMEC was increased after H/RI. The proliferation (0.36 ± 0.07 vs. 0.19 ± 0.03, P < 0.05) and migration ((55 ± 3)/5HPF vs. (28 ± 2)/5HPF, P < 0.05) abilities of CMEC were significantly improved while the apoptosis index ((9.6 ± 0.7)% vs. (19.7 ± 0.8)%, P < 0.05) was significantly decreased in H/RI+resveratrol group compared to H/RI group.SVV protein expression was also upregulated in H/RI+resveratrol group compared to H/RI group (P < 0.05). To further ascertain the role of SVV in the protective effects of resveratrol, PI3K specific inhibitor LY294002 was added to H/RI+resveratrol group, the proliferation (0.25 ± 0.05 vs. 0.36 ± 0.07, P < 0.05) and migration ((34 ± 3)/5HPF vs. (55 ± 3)/5HPF, P < 0.05) abilities were significantly decreased, the apoptosis index ((16.2 ± 0.6)% vs. (9.6 ± 0.7)%, P < 0.05) was increased and the protein expression of SVV was downregulated (P < 0.05) in LY294002+H/RI+resveratrol group compared to H/RI+resveratrol group.
CONCLUSIONResveratrol could significantly reduce H/RI induced apoptosis and attenuate H/RI induced cardiac microvascular endothelial cells dysfunction through up-regulating PI3K/Akt/SVV pathways.
Animals ; Apoptosis ; Cell Proliferation ; Chromones ; Endothelial Cells ; Enzyme Inhibitors ; pharmacology ; Heart ; Hypoxia ; Morpholines ; Myocardium ; Myocytes, Cardiac ; Phosphatidylinositol 3-Kinases ; drug effects ; metabolism ; Proto-Oncogene Proteins c-akt ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Stilbenes ; pharmacology ; Up-Regulation