| Object:To investigate the curative effect of the co-integrated vector of r AAV-PR39-ADM, which secretory expresses angiogenesis peptide(PR39) and Adrenomedullin(ADM), in the ischemia-reperfusion injury of SD rats. The ischemia-reperfusion injury models were established in SD rats. The co-integrated vector of r AAV-PR39-ADM was then injected into the injured myocardium, the functional performance was evaluated by means of echocardiography and haemodynamics methods. Finally, the expression of downstream factor was examined via molecular biology methods. Methods:1. The part 1: 36 healthy adult male SD rats, weighing 280g±20g, were randomly divided into three groups: saline(I/R), treatment group 1(TR1), treatment group 2(TR2), 12 rats in each group. The part 2: 48 healthy adult male SD rats, weighing 280g±20g, were randomly divided into four groups: treatment group 1(TR1), treatment group 2(TR2), control group 1(CG1) and control group 2(CG2), 12 rats in each group.2. After establishing ischemia-reperfusion injury model in all groups, separate treatments were carried out as follows. The part 1: For rats in control group, the saline 0.3ml was injected into rats tail vein; For rats in TR1, the co-integrated vector of r AAV-PR39-ADM 0.3ml(3×109pfu) was injected into rats tail vein; For rats in TR2, the co-integrated vector of r AAV-PR39-ADM 0.3ml(3×109pfu) was injected into localized position of the ischemia-reperfusion injury myocardium. The part 2: For rats in TR1, the co-integrated vector of r AAV-PR39-ADM 0.3ml(3×109pfu) was injected into localized position of the ischemia-reperfusion injury myocardium. In a similar way, the rats in TR2, CG1 and CG2 were injected with the co-integrated vector of r AAV-PR39-ADM 0.3ml(3×109pfu), Adeno-associated virus(AAV),0.3ml(3×109pfu) and normal saline 0.3ml(3×109pfu),respectively.3. Electrocardiogram and pathologic staining and myocardium weighing were used to monitor the injury of myocardium models in SD adult rats. After 7 days of diffident settlements, functional performance of the injured myocardium was examined in each group. Echocardiography was used to measure the left ventricular wall thickness and to calculate left ventricular ejection fraction(EF%). Haemodynamics methods were used to evaluate the left ventricular systolic pressure(LVSP)、+LV dp/dt max and-LV dp/dt max. And the myocardial fibrosis area(MIA) was evaluated by masson staining.4. For detection of associate factors, the hearts were immediately extracted after the execution of the rats, and proteins of injured area were collected and then subjected to western bolt analysis. Results:[5] Ischemia-reperfusion injury models were successfully obtained in eachgroup, as reflected by the TTC pathologic staining.[6] Injected into localized position of the ischemia-reperfusion injurymyocardium. batter than peripheral administration.3. The part2:7 days after treatment, higher index of EF%、LVSP and ±LV dp/dt max were found in TR1, as compared with TR2, CG1 and CG2. The EF%、LVSP and ±LV dp/dt max values in TR2 were also higher than CG1 and CG2, while no statistical difference was found in CG1 and CG2. The myocardial infarction area(MIA) in TR1 and TR2 groups were found smaller than those in CG1 and CG2 group. Further comparison also revealed that the MIA in TR1 group was lower than that in TR2 group, and the difference of MIA in CG1 and CG2 groups were not detectable.4. In the protein obtained from ischemia injured myocardium, the expression of hypoxia-inducible factor-1α(HIF-1α) 、 vascular endothelial growth factor(VEGF)and Adrenomedullin(ADM)were successfully detected, the content of which were analyzed, in group TR were higher than those in group CG. Conclusions:1. Ischemia-reperfusion injury model was successfully obtained in adult SD rats.2. Rats in the treatment group had enhanced function left ventricular, featuring improved contraction ability and decreased myocardial fibrosis area.3. Mechanism of improvement of heart function may be related to Adrenomedullin(ADM) and vascular endothelial growth factor(VEGF) expression in myocardium. |
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