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I. Background With the improvement of people's living standards and accelerate the aging population, the incidence of coronary heart disease and mortality rising rapidly, the composition of the Chinese residents of the fastest rising cause of death of the disease, China has become a threat to public health important diseases, which has been called \Modern medical technology, diagnosis and treatment of coronary heart disease have made great progress. Intravenous thrombolysis, percutaneous transluminal coronary angioplasty, coronary stenting technology is widely used in clinical revascularization after acute myocardial infarction mortality decreased significantly, but the ensuing myocardial ischemia - reperfusion injury (myocardial ischemia / reperfusion injury, MI / RI) has become an important factor in the patient's life threat. Therefore, to explore MI / RI study the pathogenesis and effective prevention drugs is significant. MI / RI myocardial blood supply is interrupted in a short period of time to restore the blood supply, the original blood supply to ischemic myocardium occurs before recovery compared to more serious injury. Mainly for reperfusion arrhythmias, no-reflow, myocardial stunning, microcirculation, and even sudden death and a series of conditions have worsened phenomenon. The pathological process is complex, involving multiple links. In the past for MI / RI basic and clinical research has focused on the inhibition of oxygen free radicals, antagonize intracellular calcium overload, etc., have recently considered MI / RI is primarily an inflammatory reaction, ie neutrophils (PMN) was body, platelets, endothelial cells participate in cytokines, complement, and other biologically active substances together, by the PMN adhesion molecule-mediated endothelial cell adhesion and infiltration of the myocardium and release large amounts of oxygen free radicals and enzymes category, at the cellular, subcellular and even horizontal gene cause various forms of injury. Studies have shown that, MI / RI of PMN adhesion, penetration, chemotaxis, activation and migration of endothelial wear, depends on a variety of adhesion molecules and cytokines, which are more influential tumor necrosis factor a (TNF-α), Interleukin -1 (Interleakin-1, IL-1), interleukin -6 (Interleakin-6, IL-6) and so on. These cytokines can directly inhibit myocardial contractile function, leading to decreased cardiac output, trigger apoptosis of myocardial cells directly and indirectly increase PMN adhesion molecules CD11b and endothelial cells and cardiac surface expression of intercellular adhesion molecule -1 (ICAM -1), the expression In recent years, Chinese medicine in the prevention and treatment of cardiovascular disease has made great progress. Chinese medicine although no MI / RI concept, but the etiology, clinical presentation, prognosis and other features, the scattered Chinese \. Total pathogenesis can be summarized as vacuity, the virtual person, because lack of endowment, elderly renal failure, the heart of the camp deficiency caused little yin and yang, qi and blood deficiency, especially the heart and the heart qi deficiency, and rooted in the spleen kidney; standard facts, based cream beam Atsumi, impassioned radical, labor escaping degrees, the heat generated obstruct blood stasis and qi stagnation, blood stasis, phlegm, Hanning, hot junction, especially phlegm, chest repressor Yang, occlusion of Tongxinluo, paralysis caused pain. Clinical manifestations of deficiency mixed, into each other. Treatment principles to benefit the blood, regulating yin and yang, blood stasis, phlegm polyester-based drink. Chinese herbal compound DXR (DingXin Recipe, DXR) is my school treat tachyarrhythmia clinical experience side, with a pure heart and soothe the nerves, Yiqihuoxue effect. Centering preliminary experimental study found significantly reduced order MI / RI, effectively reduce the incidence of reperfusion arrhythmias. DXR is the main drug Salvia contains a variety of active ingredients of traditional Chinese medicine, is widely used in clinical prevention and treatment of cardiovascular diseases. Its active ingredients tanshinone Ⅱ A (Tanshinone Ⅱ A) is its highest content of fat-soluble active ingredient, the damaged myocardium has a good protective effect. Our previous on DXR control MI / RI done a lot of research, but in terms of the mechanisms of inflammation dabble small. In addition, preliminary proteomics studies have shown that anti-proliferative protein (Prohibitin, PHB) in MI / RI model group and the normal group, there were significant differences in expression, and foreign research results coincide. Studies have also shown that inflammatory cytokines in intestinal epithelial cells to increase IL-6 stimulation of PHB protein and mRNA levels, and can induce activation of PHB initiation factor, but less in cardiac research. Second, the purpose of this project intends basis of previous studies using rat MI / RI animal models, from cardiac arrhythmia score and pathological observation of both traditional Chinese medicine DXR and tanshinone Ⅱ A monomer is inhibited MI / RT, play cardioprotective effect; study MI / RI serum inflammatory cytokines IL-6 and IL-6mRNA myocardial tissue expression changes and DXR and Tanshinone Ⅱ A and the effects; while the level of research in proteomics MI / RI large Mouse PHB protein expression and DXR and intervention effect of Tanshinone Ⅱ A; preliminary study in MI / RI, the inflammatory cytokines IL-6 and PHB proteins are relevant. Thus confirmed the animal level, and traditional Chinese medicine DXR monomer tanshinone Ⅱ A feasibility prevention of ischemic heart disease, to elucidate MI / RI pathogenesis, research and development of new, safe prevention of ischemic heart disease Chinese medicine provide some experimental evidence. Third, the method 32 adult male Wistar rats were randomly divided into four groups, each eight were sham group (sham operated group, SH group), myocardial ischemia - reperfusion group (myocardial ischemia-reperfusion group, MI / RI group), DXR intervention group (DingXin Recipe group, DXR group) and Tanshinone Ⅱ A intervention group (Tanshinone Ⅱ A group, Tan group). All animals were fed for 7 days. SH group, MI / RI group and Tan group were fed with normal saline, DXR DXR group fed with liquid, morning and afternoon twice intragastrically for 7 days, MI / RI group, DXR The surgical group and Tan group mold, SH groups, but not coronary artery ligation wear line, Tan preoperative intravenous injection. According to \ECG ECG machine and connect an oscilloscope to observe the incidence of arrhythmias, detailed records during reperfusion arrhythmias, ventricular arrhythmias and conduct quantitative score to evaluate the extent of reperfusion injury; abdominal aortic blood serum was separated; Take heart, ice saline, isolated left ventricle, in part directly into 10% neutral formalin fixed, some placed in -80 ℃ refrigerator. Left coronary artery ligation and reperfusion-induced arrhythmias in PVCs (ventricular extrasystole, VE), ventricular tachycardia (ventricular tachycardia, VT), VF (ventricular fibrillation, VF) and other ventricular arrhythmias (ventricular arrhythmia, VA) is Lord, it is based on the creation of VA Cutis other scoring methods for ventricular arrhythmia score. Formalin fixed paraffin-embedded myocardial tissue, sliced. Conventional HE staining myocardial tissue morphology; observation ischemia - reperfusion myocardial tissue morphological changes and DXR and tanshinone Ⅱ A on its protection. Separation of the abdominal aorta blood serum, using double antibody sandwich ABC-ELISA assay of serum IL-6 expression was observed DXR and Tanshinone Ⅱ A on ischemia - reperfusion injury in rat serum IL-6 expression. Part of the heart tissue extract total RNA, reverse transcribed cDNA, using real-time quantitative reverse transcription polymerase chain reaction (Real Time RT-PCR) detection of the expression of IL-6mRNA observe DXR and Tanshinone Ⅱ A on reperfusion injury big murine IL-6mRNA expression. Part of the heart tissue extract myocardial tissue protein, for protein blotting. And Western blot were used SABC immunohistochemistry detection of myocardial PHB protein expression was observed ischemia - reperfusion injury in rat myocardium PHB protein expression and DXR and tanshinone Ⅱ A on its expression. Arrhythmia score is a multi-level data set, using completely randomized design data rank sum test (Kruskal-Wallis) method to analyze experimental data indicate an average rank. Rest of the data is completely randomized design of the measurement data, so the use of single-factor analysis of variance (One-Way ANOVA) methods of analysis, homogeneity of variance when the Bonferroni method for multiple comparisons, when heterogeneity of variance using Dunnett's T3 method, the experimental data were expressed as mean ± standard deviation (x ± s) that the correlation between two variables were analyzed using Pearson correlation analysis. P lt; 0.05 indicates a statistically significant difference. Using SPSS13.0 software for statistical analysis. Fourth, the results of a centering parties and tanshinone Ⅱ A on ischemia - reperfusion arrhythmia. Arrhythmia score in each group showed: VA rating scores in order were: MI / RI group, Tan group, DXR groups, SH groups. The statistical test, there is a significant difference (x2 = 22.360, P = 0.000). SH rats occasional premature ventricular contractions, and no other cardiac arrhythmias; MI / RI rats were 8 cases of arrhythmia, the majority of rat coronary recanalization 1 min began to appear, with frequent PVCs, VT , ventricular fibrillation, etc, there is a small amount of grade Ⅱ and Ⅲ degree atrioventricular block, including two cases of death due to ventricular fibrillation, VA score compared with the SH group was significantly higher; DXR 8 patients were also arrhythmia, but ventricular tachycardia and ventricular premature dominated, VA score and MI / RI significantly reduced compared; Tan group type of arrhythmia also dominated PVCs and ventricular tachycardia, including one case only see occasional premature ventricular contractions, VA score and MI / RI group also significantly reduced compared. 2, DXR and Tanshinone Ⅱ A on MI / RI myocardial tissue structure has a good protective effect. HE staining light microscopy see: SH rat myocardial cells arranged in neat rows, evenly colored, membrane integrity, no degeneration, necrosis, inflammatory cell infiltration and other changes; MI / RI cardiomyocytes, disorganized, uneven coloring, some Regional myocardial cells cloudy swelling, necrosis, a large number of inflammatory cell infiltration; DXR group and Tan rat myocardial cell degeneration and necrosis have also changed, but to a lesser extent, relatively uniform coloring, degenerated reduced myocardial necrosis smaller range. 3, DXR and Tanshinone Ⅱ A on MI / RI serum IL-6 expression. Multiple comparisons were significantly different (F = 27.783, P = 0.000). MI / RI serum IL-6 levels were significantly higher than the SH group, the difference was statistically significant (P = 0.000); DXR and tanshinone Ⅱ A intervention group, serum IL-6 levels were lower than MI / RI group (P = 0.001,0.004) and higher than SH group (P = 0.001,0.000); DXR and tanshinone Ⅱ A difference between the two groups was not statistically significant (P = 1.000). 4, DXR and Tanshinone Ⅱ A on MI / RI myocardial tissue IL-6mRNA expression. Among groups myocardial tissue expression of IL-6mRNA significant difference (F = 22.499, P = 0.000). MI / RI rat tissue IL-6mRNA levels were significantly higher than the SH, the difference was statistically significant (P = 0.000); DXR and tanshinone Ⅱ A intervention group, IL-6mRNA content below MI / RI group ( P = 0.002,0.010) and higher than SH group (P = 0.002,0.000); DXR and tanshinone Ⅱ A difference between the two groups was not statistically significant (P = 1.000). 5, DXR and Tanshinone Ⅱ A on MI / RI myocardial PHB protein expression. Western blot and immunohistochemistry two methods PHB protein expression results are consistent, multiple sets of PHB expression in myocardial tissue was significantly different (F = 96.215,207.061, P = 0.000). MI / RI group of PHB expression of SH groups increased, the difference was statistically significant (P = 0.000), and tanshinone Ⅱ A DXR intervention group, PHB expression was significantly higher than the MI / RI group, the difference was statistically significant ( P = 0.000); DXR and tanshinone Ⅱ A difference between the two groups was not statistically significant (P = 1.000,0.381) V. CONCLUSIONS 1, ischemia - reperfusion injury in rats induced arrhythmias easy centering parties and tanshinone Ⅱ A can significantly reduce the incidence of arrhythmias, reduce ischemia - reperfusion injury severity. 2, DXR and Tanshinone Ⅱ A on ischemia - reperfusion injury in myocardial tissue structure has a good protective effect. 3, myocardial ischemia - reperfusion injury in rats serum IL-6 levels were significantly increased, DXR and tanshinone Ⅱ A can significantly reduce the release of IL-6, reduce reperfusion injury. 4, myocardial ischemia - reperfusion injury in rats induced expression of IL-6mRNA myocardial tissue was significantly higher DXR and tanshinone Ⅱ A can significantly reduce the expression of IL-6mRNA. 5, myocardial ischemia - reperfusion injury caused by myocardial tissue PHB protein expression increased DXR and tanshinone Ⅱ A can significantly increase the PHB protein, has a protective effect on the damaged myocardium.
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