| AimsCardiac ischemia reperfusion is that hearts lose the blood supply for a short time, the ischemia myocardium occurs a more severely injury after the blood recovered. In clinical manifestations, patients have some symptoms such as blood pressure dropped, heart dysfunction, cardiac arrhythmia and sudden death. The exact mechanisms of ischemia reperfusion injury remain unknown now, but three viewpoints about the cardiac ischemia reperfusion have been accepted by peoples, ie. calcium overload theory, oxygen free radicals theory and active leucocytes theory. To investigate the mechanism of the cardiac ischemia reperfusion is very significant for finding the efficacious ways to resolve the prevention and cure problems of the reperfusion injury. Study suggests that intracellular calcium overload plays an central role in the ischemia reperfusion.Part I: Twelve Wistar rats were randomly dividied into two groups. The control group rat hearts were perfused with Krebs for 20 minutes, ischemia for 30 minutes and reperfused with Krebs for 30 minutes. And the experimental group rat hearts were perfused with compound danshen injection for 20 minutes, ischemia for 30 minutes and reperfused with Krebs for 30 minutes. The changes of heart rate(HR), coronary flow(CF), systolic pressure(SP) and cardiac output(CO) between the control group and the experimental group can be observed. And to knowabout lactate dehydrogenase (LDH) in coronary flow and the cardiac necrotic areas between the control group and the experimental group is to evaluate the protective effects of compound danshen injection on the isolated rat hearts.PartII: After the reperfusion, the cardiac sarcoplasmic reticulum calcium(SR)were purified. Comparing SERCA Ca2+-uptake of the control group with that of the experimental group by UV spectrophotometer is to explain the possible mechanism that compound danshen injection can decrease the calcium overload .MethodsParti: (1) After rats were anaesthesia with aether, rat hearts were immediately scissored and put into the ice-cold Krebs-Henseleit butffer. And the. rat hearts were connected with the Langerdorff in a short time. The fluid of perfusion was Krebs-Henseleit with the mixture of 9 5 % oxygen and 5 % carbon dioxide. The flux from the artery and from the coronary were received separately during the perfusion. (2) The LDH in the coronary flows was observed after 30 minutes reperfusion with the UV spectrophotometer. (3)The cardiac necrotic areas could be observed by the TTC staining.Partll: The cardiac SR were purified .The changes of SERCA calcium uptake between the control group and the experimental group were examined by UV spectrophotometer. The reaction temperature maintains about 25° C, and the reaction was monitored in 710nm. Interval of sampling is 0. 1 second.The reaction time was about 100 seconds. ResultsPart I:1. comparing the change of the heat rate(HR), systolic pressure (SP), coronary flow (CF) and cardiac output (CO) between the control group and the experimental group.The HR, SP, CF and the CO between the control group and the experiment group had no significant discrimination during the 10 minutes perfusion.The HR of the twelve rat hearts was about 250~300beat/min. The SP of the 12 rat hearts ranged from the 70 to lOOmmHg. All the rat hearts accorded with the hemodynamics standards.During the 20 minutes perfusion, the values of HR, SP, CF and the CO in experimental group were all higher than the control group. Compared with the control group, in the experimental group the HR was increased, have no significant discrimination. The SP was increased, but no significant discrimination. The CF and the CO were both increased, had significant discrimination.In the 30 minutes cardiac ischemia, rat hearts were in the attemperator. The temperature remained about 37° C. 2. Comparing the change of the LDH after the reperfusion the control group with that of the experimental group.After the reperfusion, the activity of LDH in the coronary flow in the control group showed higher than that in the experimental group (136.7±13 vs 30.6 + 6.2, P<0.05).The concentrations of LDH had significantdiscrimination.3. Comparing the change of the cardiac necrotic areas between the controlgroup with that of the experimental group.After the cardiac reperfusion, the rat hearts of the control group and the experimental group were stained by the TTC. Cardiac necrotic areas appeared in both groups. The necrotic areas in the control group was larger than the experimental group(30±4. 76% vs 18 + 2. 39, P<0. 05), which had significant discrimination.Part II: Comparing the change of the SERCA calcium uptake in the control group with the experimental groupSERCA calcium uptake can be examined by the UV spectrophotometer, and the calcium uptake focused on the initiate 60 seconds . In the later time, the speed of the calcium uptake decreased slowly. With the change of the time, absorbency increased which showed the tendency of SERCA calcium uptake. The tendencies can have nonlinear regression by the SPSS 11.5. These are the regression equations and the related counts(R0, the control group: y = 15. 589Ln(x) - 3. 117 , R2 = 0.9965;the experimental group: y = 21.770Ln(x) - 3. 6841, R2 = 0.9949. The coefficient of the Ln (x) represent the slope of the curves increasing. The original speed of the calcium uptake and the half time of the calcium uptake were calculated by Excel. Compared with the control group, in the experimental group the values of the original speed of the calcium uptake increased(5. 34 ± 2.10 vs. 7.4 + 1.91, P<0. 05). The values of the original speed of thecalcium speed had the statistics significance. Conclusions1. The compound danshen injection can increase the values of CF and CO, and which had statistics significance. But the value of HR and SP had no statistics significance. The compound danshen injection can decrease the LDH releasing after cardiac ischemia reperfusion and alleviate the injury degree of the cardiac reperfusion injury.2. The compound danshen injection also can reduce the cardiac necrotic areas after ischemia reperfusion, decrease the cardiac injury degree, and then protect the isolated reperfusion rat heat.3. The compound danshen injection can increase the ability of SERCA calcium uptake, and decrease the concentration of the intracellular calcium after the cardiac ischemia reperfusion, which can alleviate the calcium overload in reperfusion injury and protect the rat heart.4. The results of this study had approved the protective effect of the compound danshen injection on the isolated ischemia reperfusion rat heart, and had more knowledge about decrease the calcium overload, which maybe help to apply new ideas about prevention and cure the cardiac ischemia reperfusion injury.
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