| Object:To observe the protection of different doses of yanhusuo total alkaloids on acute myocardial ischemia model rats, we set changes of ST and T wave of chest ECG, content of CK-MB in serum and coverage of myocardial infarction as indexes. To find the mechanism of how yanhusuo total alkaloids effect on the level of cardiac cell apoptosis, we set changes of bcl-2, bax and caspase-3 genes as indexes. This study will provide some useful data for the application of yanhusuo and can further settle a solid foundation for new medicines of yanhusuo which we have the intellectual property.Methods:Firstly, we got 70 healthy Wistar rats from Heilongjiang University of Chinese Medicine. These weights are 220±20 gram. Male or female accounts for 50% of these rats.Then we select the rats which have the healthy ECG and set them into 7 groups randomly. The 7 groups contain normal group, model group, sham operation group, yanhusuo total alkaloids high doses group, yanhusuo total alkaloids middle doses group, yanhusuo total alkaloids low doses group, positive control group. There are 10 rats in each group. The rats of normal group, model group and sham operation group were given physiological saline by gastric lavage. The yanhusuo total alkaloids high doses group, yanhusuo total alkaloids middle doses group, yanhusuo total alkaloids low doses group were given 0.5mg/kg, 1.0mg/kg,2.0mg/kg yanhusuo total alkaloids and the positive control group was given 0.5mg/kg solute of Ke da ling tablets by gastric lavage.14 days later, after the last gavage, we made the rats anesthesia injecting 20% carbamate 5ml/kg from abdomen. In order to get the model of acute myocardial ischemia, the left anterior descending coronary of hearts'artery of rats were ligated except normal group. Then we use BL-420F system to record ECG of rats and analyze the changes of HR, ST segmet, T wave in different periods. After 240 minutes, we got 2 mL blood from the artery. The serum were analyzed by semiautomatic biochemistry analyzer to acquire the changes of CK-MB. After we finished drawing blood, we separated the hearts from rats'bodies. We dealt with the hearts for tissue sections and some of these sections were fixed by formaldehyde and others were put in liquid nitrogen. The tissue sections fixed by formaldehyde will be used in TUNEL test and other sections kept in liquid nitrogen will be used for PCR. We will acquire the amount of the expression of bcl-2,bax and caspase-3 genes from PCR during acute myocardial ischemia.Results:1. Comparing model and yanhusuo total alkaloids low doses groups with normal group, there is significant difference. Comparing sham operation, yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses groups with normal groups there is no significant difference. The ST segments of model and yanhusuo total alkaloids low doses groups decrease and there is no significant discrepancy with normal group. And the ST segments of sham operation, yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses groups are similar with normal group. The T waves rise high in model groups and low doses groups and this situation is significantly different from normal group. The T waves also rise in sham operation, yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses, however, the discrepancy is not significant with normal group.2. The content of CK-MB in the serum of model group and yanhusuo total alkaloids low doses groups is significantly higher than normal group. However, the content of CK-MB in the serum of yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses groups are similar with normal group.3. In TUNEL test, the positive rate of cell apoptosis in model group and yanhusuo total alkaloids low doses groups is much higher than normal group. However, the positive rate of cell in sham operation, yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses groups is relatively low and there is no significant discrepancy.4. After PCR amplification, the expression of bax and caspase genes in model group and yanhusuo total alkaloids low doses groups is higher than normal group and bcl-2 gene is lower than normal group. However, the expression of bax and caspase-3 genes in sham operation, yanhusuo total alkaloids high doses and yanhusuo total alkaloids middle doses groups is lower than normal group and bcl-2 gene is higher than normal group.Conclusions:1. In this study, we ligate the rats'left descending coronary artery and succeed in creating the model of acute myocardial ischemia which is appropriate.2. In this study, we ligate the rats'left descending coronary artery and succeed in creating the model of acute myocardial ischemia which is appropriate.3. The high and middle yanhusuo total alkaloids can improve ST segment and T wave.4. The high and middle yanhusuo total alkaloids can decrease the content of CK-MB in the serum during the acute myocardial ischemia.5. The high and middle yanhusuo total alkaloids can protect heart when it faces myocardial ischemia and can decrease the expression of apoptosis genes.6. Although the high yanhusuo and middle total alkaloids can protect heart when it faces myocardial ischemia, the mechanism still needs exploration. |
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