The Protective Effect Of Geniposide On Cardiomyocyte Damage Under The Action Of Palmitic Acid

Objective: In this study,H9C2 cardiomyocytes were used as experimental vectors.Palmitic acid(PA)was used to simulate the high-fat model.H9C2 cells were pretreated with geniposide(Gen)to explore the protective effect and mechanism on PA-induced lipotoxicity injury in H9C2 cells.Research methods: 1.The cells were treated with different concentrations of PA,cell viability was measured with CCK-8.We select the most suitable PA concentration for modeling.The experiment grouping was as follows:control group,different concentrations of PA group(50 ??,100 ??,150 ??,200 ??,300 ??).2.The cells were treated with different concentrations of Gen,cell viability was measured with CCK-8.We select the most suitable Gen concentration.The experiment grouping was as follows:control group,different concentrations of Gen group(20 ??,40 ??,80 ??,160 ??,320 ??).3.According to the previous experimental results grouped as follows: Control group,PA group(200 ??),Gen+PA group(320 ?? Gen + 200 ?? PA,Gen group(320 ?? Gen);Whether Gen has protective effect on PA stimulate H9C2 cells were detected by CCK-8 and LDH kit.Oxidative stress were detected by DHE(ROS)staining and MDA kit.Apoptosis related proteins(Bax,Cleaved-Caspase-3,Bcl-2)and inflammasome proteins(NLRP3)and signal pathway related proteins(AKT,GSK-3?,PICK1,PKR2)were were examined by Western Blot.Results: 1.When the concentration of PA ranged from 50 ?M to 300 ?M,the viability of H9C2 cells decreased gradually.The cells survival rate was the highest when the concentration of Gen was 320 ?M.The survival rate on PA-induced lipotoxicity injury in H9C2 cells was increased,when the concentration of geniposide was 320 ?M.At the same time,the content of LDH decreased.The results showed that Gen could alleviate lipotoxicity injury in H9C2 cells and has protective effect.2.When PA-induced lipotoxicity injury in H9C2 cells,pretreatment with Gen,the fluorescence of DHE(ROS)decreased obviously and the content of MDA decreased.It indicates that the oxidative stress in the cell is weakened.3.When PA-induced lipotoxicity injury in H9C2 cells,the expression apoptosis-related proteins(Cleaved-Caspase-3 and Bax)increased,and the expression of Bcl-2 decreased.TheGen could reverse the above-mentioned changes.These results suggest that Gen can inhibit apoptosis induced by cardiomyocyte injury.4.When PA-induced lipotoxicity injury in H9C2 cells,the protein level of NLRP3 increased.After pretreated with Gen,the protein level of NLRP3 decreased.It indicates that protection from cardiomyocyte injury is related to the inhibition of inflammation.5.PA treatment can significantly decrease the expression of P-AKT and P-GSK-3?.After pretreated with Gen,it can up-regulate the protein levels of P-AKT and P-GSK-3?.It indicates that it is related to lipotoxicity injury in cardiomyocytes.6.PA treatment can inhibit the expression of PKR2 and increase the expression of PICK1.Gen can reverse the above-mentioned changes.Conclusion: 1.Gen can increase the survival rate of H9C2 cells and inhibit the activity of LDH on PA-induced lipotoxicity injury in H9C2 cardiomyocyte.It indicates that it has protective effect on cardiomyocyte injury.2.Gen can inhibit the production of ROS and decrease the content of MDA and the regulation of apoptosis-related proteins(Cleaved-caspase-3,Bax,Bcl-2)and inflammasome(NLRP3)on PA-induced lipotoxicity injury in H9C2 cardiomyocyte.It indicates that the protective effect of Gen is closely related to the inhibition of oxidative stress,apoptosis and inflammation.3.Gen could increase the proteins levels of P-AKT,P-GSK-3?,PKR2 and inhibit the expression of PICK1 protein on PA-induced lipotoxicity injury in H9C2 cardiomyocyte.It is suggested that the protective effect of Gen may be related to AKT / GSK-3? pathway,PKR2 and PICK1 proteins.