Study On The Effect Of The Traditional Chinese Formula Fufang ZhenshuTiao-zhi Recipe(FTZ) On Hyperglycemic Inducted Myocardial Cell Injury And Its Mechanism Via Antioxidation

ObjectiveThe myocardial injury induced by diabetes mellitus is one of the serious cardiovascular complications,which seriously endangers human health.The traditional Chinese formula Fufang Zhenshu Tiao-zhi recipe(FTZ)improves the myocardial injury caused by diabetes but the mechanism of FTZ is not clear.In this study,the model of myocardial injury induced by hyperglycemia was established in vitro.The effect and mechanism of traditional Chinese medicine(FTZ)on myocardial injury induced by high glucose were discussed from the perspective of oxidative stress.Methods1.Establishment of the model: H9C2 cardiomyocytes were cultured on DMEM low glucose medium and treated with glucose medium with different gradient concentrations.The survival rate of H9c2 cells was measured by MTT assay.H9C2 cardiomyocytes were divided into normal group(5.5 mmol/L),high glucose group(33 mmol/L)and mannose group(osmotic pressure compared to high glucose).The Flow cytometry was used to detect the apoptosis rate of cardiomyocytes and The Commercial kit was used to detect the formation of intracellular reactive oxygen species(ROS)and the related indexes of oxidative stress,such as malondialdehyde(MDA),catalase(catalase),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and so on.These results was used to checkout wherther the model of myocardial cell injury induced by high glucose was established.2.The effect and mechanism of FTZ: H9C2 cardiomyocytes were cultured and divided into 5 groups: normal group(5.5 mmol/L),high glucose group(33 mmol/L)FTZ low dose group(high glucose+0.2ug/m L FTZ),FTZ medium dose group(high glucose+1ug/m L FTZ),FTZ high dose group(high glucose+5ug/m L FTZ).The protein expression of apoptotic protease caspase3 was detected by WB,detection the activity of oxidative stress related indexes(MDA,CAT)of oxidative stress by Kit,the expression of genes such as NOX4,NOX2 and inflammatory factors by PCR,the expression of protein in PKC-NOX pathway such as PKC,NOX4,NOX2,i NOS,NF-k B were detected by WB.Results1.Compared with the normal group,the percentage of cell apoptosis was significantly increased,the activity of ROS in the cells was significantly increased,and the inflammation was enhanced in high glucose group.The activities of superoxide dismutase(SOD)and GSH-Px were significantly decreased in the high glucose group.2.Compared with the model group,FTZ could significantly decrease the apoptosis,and the activity of LDH and MDA and the mRNA of inflammatory factors;FTZ could increase the activity of CAT compared with the model group.3.Compared with the control group,the expression of NOX4,NOX2 protein and mRNA in the model group was significantly increased,and the expression of i NOS,PKC?2 phosphorylated NF-k B phosphorylated protein was significantly increased.Compared with the model group,FTZ could significantly reduce the expression of NOX4,NOX2 protein and mRNA,the expression of i NOS,PKC?2phosphorylated NF-k B phosphorylated protein was significantly reduced.Conclusions1.High glucose could casue the injury of cardiomyocyte.2.FTZ could improve the myocardial cell injury induced by high glucose.3.FTZ decreases the cardiomyocyte damage by inhibiting the oxidative stress and inflammatory reaction.