The Protective Effect Of Resveratrol On Cardiac Microvascular Endothelial Cells Injury Induced By Ischemia-reperfusion

Background and aimsCoronary heart disease (CHD) is one of the major diseases that threaten human health. Epidemiologic survey disclosed, French have a very low mortality rate due to CHD despite having a high-fat diet and smoking habbits. This so-called'French paradox'has been attributed in part to wine consumption, particularly red wine. Resveratrol (trans-3,4',5-trihydroxystilbene), a naturally occurring phenolic compound abundantly found in grape skins and in wines, has been thought to be responsible for the cardiovascular benefits associated with moderate wine consumption. Resveratrol has extensive physiological pharmacological effects, which include antioxygen, cleaning free radical, antiplatelet, regulating blood fat, preventing artherosclerosis, anti-inflammatory, anti-eumycete, anti-tumor, and hepatoprotective properties. The objective of this experiment is to study protective effects of resveratrol on cardiac microvascular endothelial cells (CMECs)injury induced by ischemia-reperfusion .Methods PartⅠ: The culture and identification of CMECs. 1. CMECs were isolated enzymatizally from left ventricle of SD rats, and cultured in DMEM supple- mented with 15% fetal calf serum. 2. Immunofluorescent co-stain with Dil-A c-LDL and DAPI to identify CMECs.PartⅡ: A cell model of ischemia-reperfusion injury was established. CMECs were randomly divided into three groups: Normal group (Con group), ischemia reperfusion group (I/R group) and resveratrol group (Res group). 1. Cell viability was detected by MTT colorimetry. 2. Flow cytometer based on Annexin V-FITC/PI double staining was used to detect apoptosis rates. 3. Colorimetry was used to detect caspase-3 activity. 4. The activity of superoxide dismutase and the Content of malondialdehyde were detected by ELISA. 5. The expression of TNF-a and IL-10 were detected by ELISA. 6. The positive rates of NF-кB were detected by immunofluorescence method.Statistical analysis was performed with SPSS version 11.0. One-way analysis of variation and post hoct (LSD-t) test were employed.Results1. The cells obtained showed double positive for Dil-A c-LDL and DAPI, according to the source of cells, they are CMECs .2. Compared with control group, Ischemia reperfusion injury could significantly decrease cell activity (P < 0.01); Compared with I/R group, resveratrol could significantly increase cell activity (P < 0.01).3. Compared with control group, Ischemia reperfusion injury could significantly increase cell apoptosis rates and caspase-3 activity (P < 0.01); Compared with I/R group, resveratrol could significantly decrease cell apoptosis rates and caspase-3 activity (P < 0.01).4. Compared with control group, Ischemia reperfusion injury could increase the activity of superoxide dismutase and the content of malondialdehyde. Compared with I/R group, resveratrol could significantly increase the activity of superoxide dismutase (P < 0.01), however decrease the content of malondialde- hyde (P < 0.01).5. Compared with control group, Ischemia reperfusion injury could increase the content of TNF-a and decrease the content of IL-10. Compared with I/R group, The TNF-a decreased, however the expression of IL-10 was increased.6. Compared with control group, Ischemia reperfusion injury could increase Masccline rates of NF-кB, but resveratrol could significantly decrease Masccline rates of NF-кB compared with I/R group.ConclusionResveratrol may exert protective effects by antioxygen, anti-inflammatory,decreasing CMECs apoptosis and decreasing positive rates of NF-кB.