| Objective To study whether Mongolian medicine Danggong-3 can protect myocardial ischemia-reperfusion injury by interfering mirna-126 expression from cell level,so as to provide a new drug selection and theoretical basis for clinicians to treat ischemia-reperfusion injury of coronary heart disease.Methods 1.To establish a hypoxia / reoxygenation(H / R)injury model of H9c2 cells in vitro to simulate myocardial ischemia-reperfusion(MI / R)injury.H9c2 cells were incubated with hypoxia and hypoglycemia for 3 hours,and then reoxygenated for 6 hours,as a condition of hypoxia / reoxygenation injury.2.To determine the safety and the best dosage of dangong-3.Five dosages of 0.8,0.4,0.2,0.1 and 0.05mg/ml of drugs respectively act on H9c2 cells.MTT method is used to detect the cell survival rate and determine the drugs of 0.2,0.1 and 0.05mg/ml,which respectively represent high dose,medium dose and low dose for subsequent experiments.3.Cell grouping:(1)blank control group;(2)H / R group;(3)H / R + danggong-3 high dose group;(4)H / R + danggong-3 medium dose group;(5)H / R + danggong-3 low dose group.4.The activity of AST,SOD and GSH PX were detected by ELISA.5.The expression level of mirna-126 was detected by real-time fluorescence quantitative PCR(Q-PCR).Results 1.Different dosages of danggong-3,0.8,0.4mg/ml caused H9c2 cell damage.0.2,0.1,0.05mg/ml drugs were selected for follow-up experiments.2.After H / R,the survival rate of H9c2 cells decreased to 45.79%,lower than that of the control group(P < 0.05).After drug pretreatment,high,medium and low doses could improve the cell survival rate,which were 88.55%,80.80% and 74.09%,respectively,P < 0.05,with the decrease of dose.3.After H / R,the SOD activity of H9c2 cells decreased significantly to 12.40u/mg.prot,which was lower than 21.71u/mg.prot of the control group,P < 0.05,while the high,medium and low dose of drug pretreatment could increase the SOD activity,which were 19.66,17.17,15.14u/mg.prot,higher than the model group,P < 0.05.4.After H / R,the ast release of H9c2 cells increased to 68.32u/ml,which was higher than that of the blank control group(34.78u/ml,P < 0.05).Pretreatment with dangong-3 at high,medium and low doses could reduce ast,which were 50.33,56.18,60.37u/ml,P < 0.05,respectively.5.After H / R,GSH PX activity of H9c2 cells decreased to 71.58 ? mol / L,which was lower than that of the blank control group(118.51 ? mol / L,P < 0.05);pretreatment with high,medium and low doses of danggong-3 could increase GSH PX activity,which were 104.13,93.06 and 80.30 ? mol / L,respectively(P < 0.05).6.After H / R,the expression of mirna-126 in H9c2 cells decreased to 0.42,which was lower than that in the blank control group(P < 0.05).The expression of mirna-126 was increased in three doses of dangong-3,which were 0.81,0.72 and 0.60,respectively,which were higher than that in the model group(P < 0.05).Conclusion When gong-3 pretreatment has protective effect on H9c2 cells injured by hypoxia / reoxygenation,it can reduce ast,increase the activity of SOD and GSH PX to enhance the antioxidant capacity of cells.The protective mechanism may be related to the regulation of mirna-126 expression. |
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