| Therapeutic angiogenesis in ischemic diseases has become a hot topic. The Chinese medicine of Tonifying Qi and Activating Blood has a long history and certain efficacy in treating coronary heart disease(CHD). It can improve myocardial ischemic and angiogenesis of myocardial ischemia, and it has been confirmed by a lot of clinical and experimental studies, suggesting that promoting angiogenesis may be one of important mechanisms of Chinese medicine in the treatment of CHD.This study is to probe the molecular mechanism that the Chinese medicine of Tonifying Qi and Activating Blood to promote angiogenesis in myocardial ischemia rats,to reveal the role of intervention in angiogenesis and molecular targets and provide experimental evidence for the prevention and treatment of ischemic disease.The research includes three parts:experimental research, systematic review and literature review.1. Experimental research This part includes 3 experiments:The 1st experiment:Stimulating angiogenesis effect of the Chinese medicine of Tonifying Qi and Activating Blood on the hearts of myocardial infarction rats.Objective:To study the stimulating angiogenesis effect of the Chinese medicine of Tonifying Qi and Activating Blood of Radix Ginseng and Radix Notoginseng formula on experimental myocardial infarction rats. Methods:A total of 100 Wistar rats were divided into normal control group, sham-operated group, untreated group, metoprolol (Betaloc) group, high-and low-dose Radix Ginseng and Radix Notoginseng formula groups. Model of myocardial ischemia was induced in the untreated group, the Betaloc group, the high-and low-dose Radix Ginseng and Radix Notoginseng formula groups by left anterior descending coronary artery ligation. After 12-day treatment, morphological changes of Myocardial cells were observed, serum VEGF volum was detected,microvessel density (MVD) in ischemic myocardium was detected by immunohistochemical method.Result:Radix Ginseng and Radix Notoginseng formula could reduce the myocardial cell degeneration and inflammatory cells,increase blood vessel density and serum VEGF Concentration. Conclusions:Radix Ginseng and Radix Notoginseng formula could protect ischemic myocardium, and had the roles of stimulating angiogenesis in ischemic myocardium rats after acute myocardial infarction and increasing the microvessel density in ischemic myocardium.The 2nd Experiment:Effects of the Chinese medicine of Tonifying Qi and Activating Blood on the genes expression of vascular endothelial growth factor receptor-2 and hypoxia-inducible factor-la of ischemic myocardium of rats.Objective:To investigate the effects of the Chinese medicine of Tonifying Qi and Activating Blood of Radix Ginseng and Radix Notoginseng formula on expressions of vascular endothelial growth factor receptor-2 (VEGFR-2) and hypoxia-inducible factor-la (HIF-la) in ischemic myocardium in rats with acute myocardial infarction. Methods:The methods of modeling and treating were same to the 1st experiment. After the administration, expressions of VEGFR-2 and HIF-1αproteins were detected by Western blotting, and expressions of VEGFR-2 and HIF-1αmRNAs were detected by real-time fluorescent quantitative polymerase chain reaction. Results:VEGFR-2 and HIF-la protein and mRNA expressions in the high-and low-dose Radix Ginseng and Radix Notoginseng formula groups and Betaloc were higher than those in the untreated group (P<0.05). There was a significant difference between the high-and low-dose Radix Ginseng and Radix Notoginseng formula groups (P<0.05). VEGFR-2 and HIF-1αprotein and mRNA expressions in Betaloc group were higher than the high-and low-dose Radix Ginseng and Radix Notoginseng formula groups(P<0.01). Conclusion:Radix Ginseng and Radix Notoginseng formula can up-regulate the protein and mRNA expressions of VEGFR-2 and HIF-la to improve myocardial ischemia so as to promote the development of collateral circulation in dose-dependent manner.The 3rd Experiment:Effects of the Chinese medicine of Tonifying Qi and Activating Blood on the downstream Ras signal transduction pathwayObjective:To investigate the effects of the Chinese medicine of Tonifying Qi and Activating Blood of Radix Ginseng and Radix Notoginseng formula on the downstream Ras signal transduction pathway and probe the the signal transduction pathway that the Chinese medicine of Tonifying Qi and Activating Blood promoted angiogenesis. Methods:The methods of modeling and treating were same to the 1st experiment. After the administration, expressions of Ras,c-Raf-1,ERK1/2 and p-ERK1/2 proteins were detected by Western blotting, and expressions of Ras,c-Raf-1,ERK1 and ERK2 mRNAs were detected by real-time fluorescent quantitative polymerase chain reaction. Results:ERK1/2 protein expression was not significantly different in different groups.Ras,c-Raf-1,p-ERK1/2 proteins and Ras,c-Raf-1,ERK1/2 mRNAs expressions in the high-and low-dose Radix Ginseng and Radix Notoginseng formula groups and Betaloc were higher than those in the untreated group (P<0.05). Conclusion:Radix Ginseng and Radix Notoginseng formula could up-regulate the protein expressions of Ras,c-Raf-1,p-ERK1/2and Ras,c-Raf-1,ERK1/2mRNA. It suggested Radix Ginseng and Radix Notoginseng formula possibly promote angiogenesis through Ras signal transduction pathway.2. System evaluation of clinical literature the Chinese patent medicine of Tonifying Qi and Activating Blood for Unstable Angina Pectoris:a systematic reviewObjective:To systematically analyze the effect of the Chinese patent medicine of Tonifying Qi and Activating Blood on unstable angina pectoris. Methods:①Search items were [Title:unstable angina pectoris] AND [All field:(Chinese medicine) OR (Chinese herb) OR (Integrated Traditional and Western Medicine)] AND [All field:(Tonifying Qi) OR (Activating Blood)], the search was applied to the CBM database (1994.1～2009.12).②Literature bolting was performed by 2 reviewers independently according to the standard of in and out, different opinions should be solved through negotiation.③The relation data of literature were extracted and evaluated.④A meta analysis was performed based on qualified literatures. Result:①170 literatures were found out.②Through twice blotting, there were 17 clinical study literatures that meeted the evaluate procedure.③17 literatures came into meta analysis procedures, the total number of cases were only 1644 cases.④The Chinese patent medicine of Tonifying Qi and Activating Blood concluded yu xin tong capsules, tong xin luo capsules, compound dan sen drop(tablet),shu xin yi mai capsules, bu chang wen xin granules,huo xue tong mai capsules,xue fu zhu yu capsules. Meta analysis showed that compared with the single conventional western medicine, additional use of Chinese patent medicine of Tonifying Qi and Activating Blood could significantly improve the symptoms of angina, the electrocardiogram, reduce the using amount of nitroglycerin and decrease plasma endothelin (ET), plasma a-granule membrane protein (GMP-140) which were close to the stability of atherosclerotic plaque.Conclusion:Meta analysis shows that Chinese patent medicine of Tonifying Qi and Activating Blood has some benefits in improving the symptoms of unstable angina, electrocardiogram and decreasing the using amount of nitroglycerin,ET and GMP-140. Due to lower quality of studies, the further study is need.3. Literature review123 literatures were read, collected and summarized, and progress on studies of the relationship among VEGF,CD105 and angiogenesis was explored and reviewed.A preliminary summary was conducted on literatures about Chinese medicine of Tonifying Qi and Activating Blood stimulating angiogenesis of coronary heart disease. We put forward that clinical studies combined with researches in Vitro and Vivo be conducted through many ways, many links to explore the role and signal transduction mechanisms through which chinese medicine stimulates angiogenesis of myocardial ischemia of coronary heart disease.The research choosed the Ras protein signaling pathway as the main line, and observed the intervention of Radix Ginseng and Radix Notoginseng formula on angiogenesis in Ras signal transduction system in rats and its mechanism.It is a original research on the field of Chinese medicine's prevention and treatment of coronary heart disease.Borrowing CD105 method of detection of tumor micro vessel density, the research adopted CD105 to detect micro vessel density of myocardial ischemia.It is a innovative application of the examination index. |
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