Proteomic Study On Cardioprotective Effects Of Qi-Shen-Yi-Qi Formula In Acute Myocardial Ischemic Rats

Proteomics is a new emerging technique, which offers a systematical view on the variation of expression and function of protein under certain stimuli. It helps people get new insight of drug action on the proteo-level. The traditional Chinese medicine has the characteristic of the multi-component, multi-targets and synergistic effects. Using the proteomic technique to identify the potential mechanism of traditional Chinese medicine will contribute to understand the way that the traditional Chinese medicine works by. It will also benefit to reveal modern biological meaning of the theory of traditional Chinese medicine on the molecular level. In this thesis, cardiac proteome of the acute myocardial ischemic rats were studied. A two dimensional electrophoresis (2-DE) based proteomic analytical method was developed by a series of comparative experiments. Subsequently, the cardioprotective effects of Qi-Shen-Yi-Qi formula in- acute myocardial ischemic rats were studied by the routine pharmacological assays and cardiac proteome analysis. The main research contents were as followed:1,Firstly, a 2-DE based method for the analysis of rat myocardial proteome was established. The tissue of cardiac muscle were obtained from the rats with acute myocardial ischemia. The method of myocardial proteomic sample preparation (include protein extraction and purification) was developed. Several key parameters in 2-DE analysis were optimized, including sample amount, IPG strips, voltage for focusing and SDS-PAGE. The established method was applied in analyze proteome of ischemic hearts being treated with verapamil. 15 differential proteins were well separated. The results indicated the present method was reliable and stable, which can be used to characterize myocardial proteome.2,The proteomic study on cardioprotective effects of Qi-Shen-Yi-Qi formula in acute myocardial ischemic rats. The pharmacological assays were performed to determine serum enzyme levels in each experimental group. The proteomic analysis was also progressed by the built 2-DE method to rats cardiac proteome which was treated with Qi-Shen-Yi-Qi formula for 3 and 7 days after heart ischemia. The result showed that the expression of 35 proteins were changed significantly. Using MALDI-TOF-MS, these 35 proteins were identified by database searching. These proteins can be related to energy metabolism and mitochondrial function, anti-oxidative stress property, and so on. The findings showed that the myocardial protective effects of the qishenyiqi formula in myocardial damage process are related to the recovery of energy supply as well as anti-oxidative stress property.In this thesis, the method of myocardial proteome was built successfully, and the proteomic research on the anti-ischemic effect of the Qi-Shen-Yi-Qi formula was progressed by this built method, the result afforded some clues in multiple targets mechanism of the traditional Chinese medicine.