Protective Effects And Effects On Alteration Of Protein Expression Pattern Of Potentilla Anserina L. Alcohol Extract Following Acute Myocardial Ischemia In Rats

Objective: We researched the protective effects and its Effects on Alteration of Protein Expression Pattern of Potentilla anserina L. alcohol extract following acute myocardial ischemia in Rats.Methods: Male Sprague-Dawley rats were randomly divided into control group and injury groups which were subjected to coronary occlusion lasting for 30min and reperfusion for 120min and then subdivided into model and diltiazem groups, intervention group of Potentilla anserina L. alcohol extract(0.9g?kg-1,1.8g?kg-1,3.6g?kg-1). Retrograde inserting into the left ventrical through a cannula and fixed in the right common carotid arter, recording the indexs of haemonamics. Activities of LDH, CK in serum were measured. Observe the change of mitochondrion and glycogen in myocardial cell by electron microscope. apoptotic myocardium was Semi-quantitative analyzed by TUNEL staining. The possible mechanisms on acute myocardial ischemia injury in mice were approached by biochemical indicators (SOD,MDA,NO), immuno-histochemistry stain (iNOS,Caspase3,Caspase9) and RT-PCR(caspase9,caspase3). The change of protein expression pattern in serum was monitored with weak cationic exchanger chips(CM10) and metallic anion exchange chips(IMAC3-Cu) by surface-enhanced laser desorption ionization-time of flight-mass spectrometry.Results: 1 The effect of Potentilla anserine L on acute myocardial ischemia and reperfusion injury in rats (1) Intervention group of Potentilla anserine L. alcohol extract(0.9g/kg,1.8g/kg,3.6g/kg)could significantly improved homodynamic myocardial functional parameters after(±dp/dt max) I/R injury; (2)HE staining: In all of the model group, the concentration group Effect of Potentilla anserine and the positive drug group, the scope of the injury mainly happen in the left ventricular. Model group, which histopathological changes significantly, inflammatory cells infilter in myocardial interstitial, acidophilia myocardial fibers change, sarcomere fault, disappeared necrosis obviously. Effect of Potentilla anserine high and middle dose group significantly reduced the injury compared with model group. Effect of low-dose group is not good, but there is still more to improve than the model group. (3)Myocardial ultra-structure changes: electron microscopy showed that the treatment group concentration Effect of Potentilla anserine to reduce myocardial ischemia-reperfusion injury caused by swelling of mitochondria, vacuoles, such as changes in the ultra-structure of injury. (4) Serum Biochemical Indicators: the treatment group concentration Effect of Potentilla anserine could decreased the activities of LDH(P<0.01), CK(P<0.01).(5) TUNEL staining: the concentration Effect of Potentilla anserine treatment group (compared with the model group) can be reduced significantly decreased cardiomyocyte apoptosis index (P <0.01).2 Effect of Potentilla anserine ethanol extract on rats with acute myocardial ischemia and reperfusion injury in the role of mechanisms of protection: (1) Serum MDA, SOD, NO detection: the concentration Effect of Potentilla anserine treatment group (compared with the model group) can be significantly enhanced SOD activity in rats serum (P<0.01), reduced the production of MDA (P<0.01) and NO content and lower (P<0.01,P<0.05); (2) immuno-histochemical staining: Effect of Potentilla anserine administration of each dose group (compared with the model group) can significantly reduce iNOS (P<0.01), and caspase3 (P<0.01),caspase9 (P<0.01) protein expression after ischemia-reperfusion injury. (3)RT-PCR: the high and intermediate doses of Potentilla anserine L could significantly depress the express level of caspase-9 mRAN on hypoxia injury neuron compared with model group (P<0.05), There is no significant difference between low dose of Potentilla anserina L. and model group (P>0.05). The high, intermediate and low doses of Potentilla anserine L could depress the express levels of caspase-3 mRAN on hypoxia injury neuron compared with model group (the high and intermediate doses P<0.01, the low dose P<0.05). 3 Changes of protein expression patterns: Compared with model group, there are 1 proteins higher expressed and 5 protein lower expressed in IMAC-3 of intervention group of Potentilla anserine L. alcohol extract(0.9g?kg-1,1.8g?kg-1,3.6 g?kg-1), there are 9 proteins higher expressed and 1 protein lower expressed in CM10 of intervention group of Potentilla anserine L. alcohol extract(0.9g?kg-1,1.8g?kg-1,3.6 g?kg-1).Conclusion: 1 Potentilla anserina L. alcohol extract could significantly protect acute myocardial ischemia-reperfusion induced myocardial injury, the myocardial tissue ultra-structure, and improve cardiac function; Effect of Potentilla anserine ethanol extract could significantly inhibit myocardial ischemia-reperfusion injury induced apoptosis. 2 Effect of Potentilla anserine of myocardial ischemia-reperfusion injury may be related to the protective effect of scavenging oxygen free radicals, reducing NO content, inhibited caspase family of apoptosis-related protein expression, etc 3 Myocardial injury can cause serum protein expression profile changes.Protein expression profiles are different in different treatment groups, suggesting that the serum protein may be some differences in myocardial ischemic and reperfusion injury-induced gene expression products and drug targets.