Experimental study of miRNA-210 modified MSCs in the treatment of myocardial ischemia-reperfusion injury in rats

Objective:To investigate the therapeutic effect of microRNA210(miRNA-210)modified mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:One SD rat was sacrificed,and the lower extremity tibia and femur were isolated.MSCs were cultured by whole bone marrow adherence method to construct miRNA-210 modified MSCs.40 SD rats were divided into the sham operation group,model group,MSCs group,and miRNA-210+MSCs group,with 10 rats in each group.The left anterior descending coronary artery was ligated to prepare a model of myocardial ischemia and reperfusion.After successful modeling,50μl of MSCs suspension was injected into the tail vein of the MSCs group,and 50μl of miRNA-210 modified MSCs suspension was injected into the tail vein of the miRNA-210+MSCs group.The sham operation group and the model group were injected with the same amount of normal saline.On the 10th day after modeling,the area of myocardial infarction,morphological changes of myocardial tissue,myocardial cell apoptosis rate,and miRNA-210 expression were compared in each group.Results:The area of myocardial infarction and the rate of myocardial cell apoptosis in the model group were significantly higher than those in the sham operation group(<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the MSCs group were significantly lower than those in the sham operation group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly higher than those in the MSCs group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly lower than those in the sham operation group(P<0.05).The expression level of miRNA-210 in the myocardial tissue of the model group was significantly higher than that in the sham operation group(P<0.05);There were no significantly different in the expression level of miRNA-210 in the myocardial tissue between the MSCs group and model group(P>0.05);The expression level of miRNA-210 in the myocardial tissue of MSCs group was significantly higher than in the MSCs group,model group and sham operation group(P<0.05).HE staining showed that the miRNA-210+MSCs group had normal morphology of myocardial tissues,more uniform cytoplasmic staining,and arranged neatly myocardial fibers.The inflammatory cell infiltration and interstitial edema of the miRNA-210+MSCs group were significantly improved compared with the model group and MSCs group.Conclusion:MiRNA-210 modified MSCs can inhibit myocardial cell apoptosis in myocardial ischemia-reperfusion injury model rats,reduce the area of myocardial infarction,and improve pathological damage of myocardial tissue in rats,which has a certain therapeutic effect on myocardial ischemia-reperfusion injury.

Non-invasive assesment of myocardial risk and infarct area in canine model of myocardial reperfusion by intravenous contrast echocardiography

This work is supported by Medical Science Technique Foundation of Guangdong Province.Abstract Objective To evaluate the newly developed perfluoropropene filled echo contrast agent (FCT 188) in non invasive assessment of risk areas (RA) and infarct areas (IA) with intravenous myocardial contrast echocardiography (MCE) in canine model of ischemia followed by reperfusion. Methods Eight chest opened Beagle dogs with a 90 minute ischemia followed by a 240 minute reperfusion were studied. MCE was performed after a bolus injection of FCT 188 (0.025 ml/kg, Ⅳ) into a superficial vein of the forelimb at baseline, 20 minutes after occlusion, and 4 h after reperfusion to non invasively assess the left ventricular myocardium area (LVMA), myocardial ischemic risk area (RA), and infarct area (IA) in a short axis view of left ventricle. The accuracy of detecting myocardial perfusion with intravenous MCE was further assessed by in vitro myocardial staining of the matched cross sections. Both RA and IA were expressed as percent of LVMA. Results LVMA, RA, IA, and IA/RA ratio were accurately assessed by MCE (LVMA: 6.60 cm 2±0.76 cm 2; RA: 35.7%±6.68%; IA: 21.0%±13.2%; IA/RA: 60.3%±31.4%; n=7) as compared with those of the matched cross section (LVMA: 6.81 cm 2±0.73 cm 2, P=0.062; RA: 35.3%±9.9%, P= 0.84; IA: 25.10%±14.5%, P=0.07; IA/RA: 68.0%±22.2%, P=0.28, respectively). There was a significant correlation of MCE assessed IA/RA ratio and its corresponding pathologiclly determined finding in vitro (Y=1.21X-21.6, r=0.73, P=0.015). No significant changes of electrocardiogram (ECG), mean artery pressures (MAP), pulmonary artery pressures (PAP), and pulmonary artery wedge pressures (PAWP) were found between pre and post intravenous injection of FCT 188 at each time point. Conclusion These indicate that FCT 188 can be used to assess risk areas and infarct areas accurately and non invasively with intravenous MCE in the canine model of a 90 minute ischemia followed by a 240 minute reperfusion and might have potential significance for non invasive assessment of myocardial reperfusion clinically.