| Background: Ischemic heart disease is a serious cardiovascular disease, itthreatens human health, and the pathogenesis is closely linked with complex andmultiple systems. In cellular and molecular levels, ischemia and hypoxia could affectthe energy metabolism, metabolism, and even gene expressions of myocardial cells，all these changes simultaneously contribute to the development process of myocardialischemic disease. Traditional Chinese medicine is profound, in which a number ofsubstances are proved to protect against myocardial ischemia and hypoxia, therefore,it is to find the efficacy of the active material composition in the natural medicine isthe starting point of this stage of the development of new drugs.Objective: Tongmai dixian pill is a traditional Chinese medicine, Astragalus,Eucommia, polygonum multiflorum, Cyathula Chuanxiong, Salvia, earthworm andother components with anti-lipid peroxidation, inhibition of the inflammatoryresponse, change blood rheology science relatedindicators and improve themicrocirculation of the function has been applied to clinical treatment and has a goodeffect. However, so far, there is no clear study to confirm the role of the of Tongmaidixian pill in the protection of ischemic myocardium by promoting the angiogenesisof ischemic myocardium, especially from the molecular level to study the protectiveeffects of specific angiogenic mechanism. So our study is designed to combine thewhole animal coronary artery ligation-induced injury by hypoxia and the molecularlevel from the different levels on the whole, tissue, cell and molecular study ofTongmai dixian pill protective effects on ischemic myocardium and to explore itsunderlying mechanism.Methods: Rats were anesthetised with diethyl ether and with back fixed on theoperating table, after skin preparation and sterilization, using purse-string suture of 10th line, cut the skin preparation district middle with a scalpel, dissect the intercostalmuscles in the leftside3to4intercostal thoracotomy to expose the heart, ligatedbetween the left atrium and pulmonary artery cone under23mm at the left anteriordescending artery ligation with suture of the0-line, then return the heart back into thechest, squeezed intrathoracic gas quickly and tighten the purse-string suture line, thenclose the chest, make sure the whole process was within less than30s, in the shamoperation group, only the surgical line was to play a loose knot.24h later, ECG wasdetected and model was built successfully by observing the pale color myocardium、weakened pulse and ECG ST segment elevation. Rats were randomly divided into sixgroups, sham group, model group, Tongmai dixian high, medium and low (6.30g kg-1,3.15g kg-1,1.57g kg-1) dose groups, and positive control heart-protectingpills (0.01g kg-1) group. Drug was administered once daily and weight wasmeasured once a week. Rats were sacrificed after6weeks.1.using rat ligation of the left anterior descending coronary artery preparation ofmyocardial ischemia model, measuring ST segment elevation as well as TTC stainingdetection of myocardial infarct size, research effect of the ischemia myocardial infarctsize.2.By measuring SOD, MDA concent in the serum,to research Tongmai dixian pillantioxidant against free radicals, but also reflect its ability to clear the lipidperoxidation metabolite. Serum enzymatic indicators, such as LDH, CK and AST, itcan react the extent of myocardial tissue injury and repair.3. Mesuring the extent of myocardial tissue recovery organization morphology detecttongmai to penny pill role and immunohistochemical staining of vascular growthfactor of VEGF in bFGF protein expression and CD34mark microvessel counts in theedge of the blood vessels of ischemic myocardial tissue regeneration status.4. Testing the relevant factors of the infarct border zone angiogenesis in ischemicmyocardial tissue by RT-PCR, revealing the vascular growth factor VEGF and bFGFin angiogenesis played an important role in it. Results:1. Tongmai dixian pill on the rat ECG ST-segment on S-T segment elevation andMyocardial infarction area.it can decease ST segment elevation,and compared withthe model group has statistically significant, and deduce the scope of myocardialinfarctions as to protect the heart.2. Effects of Tong Mai Di Xian Pill on SOD，MDA in serum in rats,it can have SODactivity increased, decreased MDA, proved that the drug has anti-lipid oxidation. Atthe same time, can reduce CK, LDH and AST content in the serum, play a role in theprotection of myocardial cells.3. Effect of Tong Mai Di Xian Pill postconditioning on histomorphology change ofcardiac muscle damaged by Myocardial ischemia under the inverted microscope.under the light microscope observation, cardiac muscle fibers arranged in neat rows ofthe sham operation group, the myocardial interstitial inflammatory cell infiltration;swelling model of myocardial fibers, arranged in disorders, myocardial interstitialinflammatory cell infiltration, or even visible focal necrosis; Tongmai dixian eachdose group myocardial nucleus size uniform, mild swelling of the myocardial fibers,showing varying degrees of fibroblasts and inflammatory cell infiltration; positivecontrol drug heart-protecting’s effect is similar to Tongmai dixian Middle dose group.And Immunohistochemical staining showed that after administration of vascularendothelial growth factor VEGF, bFGF protein expression is increasing, seeing theexpression of a different degree of staining area. At the same time, the detection of theCD34marker of microvascular regeneration after administration of each groupshowed a rising trend of microvessel density.4. VEGF mRNA and bFGF mRNA detected by RT-PCR technology in the ischemicmyocardial tissue is upregulated, indicating that the key factor to promote the role ofangiogenesis in the ischemic myocardial tissue may be of VEGF, bFGF, and play animportant role in the myocardial protection.According to the results above, we found that Tongmai dixian pill could protectagainst myocardial ischemia in the following mechanisms:（1） It enhanced the abilities of cardiomyocytes in anti-lipid peroxidation, reduced reactive oxygen speciesand lipid metabolites damage in myocardial cells.（2） it increased gene expression ofvascular endothelial growth factor of VEGF and bFGF, as well as increasedmicrovessel density of the myocardial infarct border zone. So we come to theconclusion that it plays an important role to protect myocardial tissues and cellsagainst ischmic injury. |
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