Ischemic Myocardial Apoptosis Signal Transduction Mechanisms And Therapeutic Oral Liquid Intervention Sirt1 Regulation Research

BackgroundIschemic heart disease (IHD) is a cardiovascular disease accompanied with decreased number of functional myocardial cells which leads to apoptosis and necrosis of myocardial cells and finally deterioration and insufficiency of heart function. The life span of myocardial cells is related to the treatment effects of ischemic heart disease and the significance of reducing apoptosis of myocardial cells is paramount. SIRT1is a homolog of mammalian yeast chroma tin silencer, Sir2, which is a NAD+dependent histone deacetylase. SIRT1is involved in the regulations of many biological processes, including multiple genes transcription, energy metabolism, cell aging and apoptosis. SIRT1is expected to be a new target for the treatment of IHD.ObjectiveTo observe the changes of H9c2rat myocardial model cells under microscopy following intervention with xuefuzhuyu oral liquid (Oxygen-glucose deprivation) and transfection with SIRT1, as well as changes of Sirtl pathway related genes (FOXO fami ly, NF-kB, p53et ct) and cell activities, so as to clarify the gene target and corresponding mechanism of xufuzhuyu oral liquid in the treatment of IHD under the TCM theory of eliminating blood stasis and promoting tissue regeneration.MethodH9c2rat myocardial cells were divided into six groups:Normal control, OGD model group, SIRT1transfection group,OGD+SIRT1transfection group, OGD+Xuefuzhuyu oral liquid treated group,OGD+SIRT1transfection+Xuefuzhuyu oral liquid treated group. CCK-8kit was used to draw cell growth curve. The changes of cell morphology were observed with light microscope after AO/EB staining. Quantitative fluorescent PCR and Western-blot were used in the determination of concentration variations of SIRT1pathway related genes and corresponding protein expression status after Xueyuzhuyu oral liquid intervention and SIRT1transfection.Results1. The result of the light microscope:in morphology, it had significantly different between the6different groups.2.RT-PCR results showed that SIRT1mRNA that copy numbers and corresponding protein expression decreased in OGD model group, SIRT1transfection group, OGD+SIRT1transfection group, OGD+Xuefuzhuyu oral liquid treated group, OGD+SIRT1transfection+Xuefuzhuyu oral liquid treated group, but increased after intervention; The expression of copy numbers and corresponding protein of SIRT1mRNA get similar in Normal control and OGD+Xuefuzhuyu oral liquid treated group,but showed significant differences between Normal control and OGD+SIRT1transfection+Xuefuzhuyu oral liquid treated group. mRNA copy numbers and corresponding protein expression of P53、NF-kB、FOXO1、FOXO3、FOXO4increased significantly in OGD model group,SIRT1transfection group, OGD+SIRT1transfection group, OGD+Xuefuzhuyu oral liquid treated group, OGD+SIRT1transfection+Xuefuzhuyu oral liquid treated group.3. We stern blot results showed that SIRT1mRNA that copy numbers and corresponding protein expression decreased in model group, but increased after intervention; mRNA copy numbers and corresponding protein expression of p53、NF-kB、FOXO1、 FOXO3、FOXO4increased in model group; Protein expression of SIRT1、p53、NF-kB、 FOXO1、FOXO3、FOXO4showed significant differences(p<0.05).ConclusionOur study proves that SIRT1signal pathway has antiapoptotic effects in in vitro OGD rat myocardial cells which reveals that SIRT1signal pathway might be one of the targets of Xuefuzhuyu oral liquid in fighting against myocardial cell apoptosis by modulating functions of its substrates and downstream genes. This study promotes the application research of Xuefuzhuyu oral liquid in cardiac regeneration medicine and provides a clear target for ischemic heart disease treatment.