| Objective: The treatment of reperfusion can save the myocardial cell in danger, but at the same time it will cause new injury, which is myocardial ischemia reperfusion injury. Recent studies have shown that myocardial cell apoptosis is an important form of death and play an important role in the pathogenesis of myocardial ischemia reperfusion injury, which can effect the development of the infarction. Rho-kinase mediated cell apoptosis.In this study, the model of ischemia /reperfusion in rats was built. Myocardial cell apoptosis and the expression of Bcl-2 and Bax were obtained, so as to explore the protective effects of different doses of Rho-kinase-inhibitor (Fasudil) on myocardial ischemia reperfusion injury.Methods: Healthy male SD rats were used in this study. After raised one week, they were randomly divided into 5 groups, Sham group(Sham): Through the line across the left anterior descending, 30 minutes later, Physiological saline (0.5ml) was injected into the peritoneum, and observed for 165 minutes; Ischemia/reperfusion group(I/R): Tighten up the line of ligature, the left anterior descending was occluded for 45 minutes. Physiological saline (0.5ml) was injected into the peritoneum 15 minutes before the left anterior descending was reperfused, then relaxed the line of ligature and reperfused for 120 minutes; High dose Fasudil treated group(HDF): Tighten up the line of ligature, the left anterior descending was occluded for 45 minutes. Fasudil (30mg/kg) was injected into the peritoneum 15 minutes before the left anterior descending was reperfused, then relaxed the line of ligature and reperfused for 120 minutes; Intermediate dose Fasudil treated group(IDF): Tighten up the line of ligature, the left anterior descending was occluded for 45 minutes. Fasudil (10mg/kg) was injected into the peritoneum 15 minutes before the left anterior descending was reperfused, then relaxed the line of ligature and reperfused for 120 minutes; Low dose Fasudil treated group(LDF): Tighten up the line of ligature, the left anterior descending was occluded for 45 minutes. Fasudil (5mg/kg) was injected into the peritoneum 15 minutes before the left anterior descending was reperfused, then relaxed the line of ligature and reperfused for 120 minutes. The arteries were obstructed once more after 120 minutes' reperfusion. The ischemia areas were determined by Evans-blue and TTC double staining. The myocardial cell apoptosis in the ischemia region was determined by TUNEL assay and the apoptotic index (AI) were calculated. The Bcl-2 and Bax protein expressions were measured by immunohistochemical technique.Results: 1 Morphological changes of myocardial tissues. In the sham group the cardiac muscle fibers lined up regularly, the structure was clear, nuclear was ellipse, and the interspaces between myocardial cells were tight, and the morphological character was almost normal. The change of I/R group in myocardium muscle tissue pathomorphology was obviously aggravated than HDF treated group, IDF treated group and LDF treated group. The change of HDF treated group was less than IDF treated group and LDF treated group. The change of IDF treated group was as LDF treated group.2 The myocardial cell apoptosis determined by TUNEL assay. Compared with the Sham group, the myocardial cell apoptosis index was obviously increased in I/R group, LDF group, IDF group and HDF group ( P <0.01);Compared with the I/R group, the myocardial cell apoptotic index was obviously reduced in LDF group, IDF group and HDF group ( P <0.01);Compared with the HDF group, the myocardial cell apoptotic index was obviously increased in LDF group and IDF group(P<0.01). There was no significant difference between LDF group and IDF group(P >0.05).3 The expression of Bcl-2. Compared with the Sham group, the expression of Bcl-2 was obviously reduced in I/R group, LDF group, IDF group and HDF group(P<0.01);Compared with the I/R group, the expression of Bcl-2 was increased obviously in HDF group, IDF group LDF group ( P <0.01);Compared with the HDF group, the expression of Bcl-2 was decreased obviously in IDF group and LDF group,(P <0.01);There was no significant difference between LDF group and IDF group(P >0.05).4 The expression of Bax. Compared with the Sham group, the expression of Bax was increased obviously in I/R group, LDF group, IDF group(P <0.01);Compared with the Sham group, the expression of Bax was obviously increased in HDF group (P<0.05);Compared with the I/R group, the expression of Bax was reduced obviously in LDF group, IDF group and HDF group(P <0.01);Compared with the HDF group, the expression of Bax was increased obviously in IDF group and LDF group(P<0.01). There was no significant difference between LDF group and IDF group(P >0.05).Conclusion: Different Doses Fasudil were treated during ischemia /reperfusion in rats. The results were as follows(1) increase the expression of Bcl-2.(2) reduce the expression of Bax.(3) reduce the myocardial cell apoptosis. The protective effects depended on the dose of Fasudil, the high dose had a better effects. In conclusion, Fasudil can protect the myocardial cell from being injured during the myocardial ischemia/reperfusion, it may be interrelated with reducing Bax expression and increasing Bcl-2 expression. |
PDF Full Text Request