Study On Protective Effects Of Shengmai Formula And Ginsenoside Rg₃ Against Chronic Heart Failure By MiRNA Expression Profiling

Shengmai Formula ?SMF? is clinically used for treatment of cardiovascular diseases, such as coronary heart disease, arrhythmia, and chronic heart failure ?CHF?. ?S?-ginsenosides Rg₃ is one of the active compounds in SMF, and its mechanism of action in myocardial protection remains to be further study. This thesis applied miRNA array technology to investigate the protective effects of SMF and ?S?-ginsenosides Rg₃ against CHF. Differentially expressed miRNAs were identified to predict the potential targets and regulated signaling pathways. Cardioprotective effects of SMF and ?S?-ginsenosides Rg₃ were validated in cellular models. Our findings provide scientific evidences for illustrating the mechanism of action of SMF.The main contents of the thesis included:1. miRNA expression profiling technology was applied to study anti-CHF effect of SMF.16 differential miRNAs were found, including 10 up-regulated and 6 down regulated miRNAs. miR-21-3p, miR-294 and miR-542-3p, which were related to myocardial hypertrophy or cell apoptosis, were verified by RT-PCR. Target gene network of differential miRNAs was established and potential regulated signaling pathways were predicted. The cardioprotective effects against hypertrophy and apoptosis of SMF were validated in Ang?-induced hypertrophy cellular model and t-BHP injuryed cellular model.2. miRNA expression profiling technology was applied to study anti-CHF effect of ?S?-ginsenosides Rg₃, an effective substance in SMF.14 differential miRNAs were found, including 8 up-regulated and 6 down regulated miRNAs. Among these differential miRNAs, miR-200-5p was found to participate in regulation of SIRT1, and SIRT1 was further researched as a target. The specific probe of SIRT1 activity was used to screen bioactive compounds in SMF.12 compounds were found as SIRT1 activators, including 7 from Panax ginseng,2 from Ophiopogon japonicas, and 3 from Schisandra chinensis. The structural characteristics of SIRT1 activitors from Panax ginseng were summarized. It was found that ginsenosides with protopanoxadiol nucleus were more likely to activite SIRT1 and S configuration of the 20 carbon played the key role. As the representative compound, pharmacological effects of ?S?-ginsenosides Rg₃ were further investigated. By molecular docking technology, it was found that ?S?-ginsenosides Rg₃ served as a link between substrate peptide and SIRT1 interaction, thus enhancing the capacity of protein and substrate combination. Results of liquid chromatography-mass spectrometry and surface plasmon resonance verified the activation of SIRT1 by ?S?-ginsenosides Rg₃. Moreover, the cardioprotective effects of ?S?-ginsenosides Rg₃ were confirmed in Ang?-induced hypertrophy cellular model and t-BHP injuryed cellular model.In summary, the thesis investigated protective effects of SMF and ?S?-ginsenosides Rg₃ against chronic heart failure by miRNA expression profiling. ?S?-ginsenosides Rg₃, which was found as a SIRT1 activator, exerted cardioprotective function. These findings provided new evidence for the mechanism of action of SMF and its bioactive compounds for treating CHF.