| Cardiac arrest is the sudden termination of heart beat,causing ischemia,hypoxia and dysfunction of the whole body organs,and is a common clinical emergency event.Although CPR technology continues to improve,the mortality rate from cardiac arrest remains high.Cardiac insufficiency after resuscitation is the main cause of early death after spontaneous circulation is restored.Therefore,how to restore the pumping function of cardiac arrest and improve the degree of myocardial damage after resuscitation is a key factor in reducing mortality after cardiopulmonary resuscitation.Shenfu injection is a common Chinese patent medicine used in clinical rescue,which has the effect of benefiting the temperature and yang,and regaining the yang to save adversity.In recent years,a large number of basic experiments have confirmed that Shenfu injection has obvious curative effect on the treatment of ischemiareperfusion injury.Therefore,this study through animal experiments and biological analysis of its tissues proves that Shenfu injection can improve asphyxia combined with ice potassium chloride The main research contents and experimental results of myocardial injury after resuscitation in induced cardiac arrest rats are as follows:Objective: By observing the regulating effect of Shenfu injection on the Nrf2/HO-1 axis of oxidative stress-related pathways,to explore the mechanism of Shenfu injection on improving myocardial injury in rats after resuscitation at the molecular level.Methods: 40 male SD rats,30 rats were randomly selected for asphyxiation combined with ice potassium chloride to induce cardiac arrest,and a series of treatment measures such as cardiopulmonary resuscitation were given five minutes later,and they were randomly divided into three groups after the spontaneous circulation was restored,Each group of 10: normal resuscitation group received 5 injections of normal saline 1ml/time 1h after successful resuscitation and at nine in the morning and five in the afternoon;Shenfu 2ml/kg group: injected Shenfu injection 2ml/kg 1h after successful resuscitation,Inject Shenfu injection2ml/kg at nine in the morning and five in the afternoon;Shenfu 7ml/kg group: Inject 7ml/kg of Shenfu injection 1 hour after successful resuscitation,and inject Shenfu injection 7ml/kg at nine in the morning and five in the afternoon.The other 10 were treated as a sham operation group: only tracheal intubation and arteriovenous catheterization were performed,and 1 ml of normal saline was injected 1 hour after the operation,and at 9 am and 5 pm.Samples were taken 24 hours later to detect serum creatine kinase isoenzyme(CK-MB)and cardiac troponin I(cTnI)levels to analyze the degree of myocardial damage after cardiac arrest and cardiopulmonary resuscitation;TBA method to detect oxygen free radical malondialdehyde(MDA))Level,WST-1 method to detect the activity of superoxide dismutase(SOD)in myocardial tissue,to determine the level of myocardial oxidative stress after cardiac arrest;Western Blot to detect the expression level of Nrf2,p-Nrf2,HO-1 protein,real-time fluorescence quantification Analysis(RT-PCR)was used to detect the expression of HO-1 m RNA and NQO1 m RNA;immunofluorescence was used to observe the fluorescent expression of pNrf2 protein in the nucleus of myocardial tissue.Results: 1.Comparison of serum myocardial enzymes showed that 24 hours after successful resuscitation,the level of CK-MB in the normal resuscitation group was higher than that of the sham operation(P<0.01),and the cTnI level of the normal resuscitation group was higher than that of the sham operation group(P<0.01);see Compared with the normal resuscitation group,the CK-MB level of the 2ml/kg injection group was lower(P=0.03),and the cTnI level of the Shenfu injection 2ml/kg group was lower than the normal resuscitation group(P=0.001);Shenfu injection Compared with the Shenfu injection 2ml/kg group,the CK-MB level of the 7ml/kg group was lower(P=0.011),and the cTnI level of the Shenfu injection 7ml/kg group was lower than that of the Shenfu injection 2ml/kg group(P= 0.039).2.The oxidative stress test results showed that the SOD activity of the normal resuscitation group was lower than that of the sham operation group(P<0.01),the 2ml/kg SOD activity of Shenfu injection was higher than that of the normal resuscitation group(P=0.023),and Shenfu injection The SOD activity of the 7ml/kg group was higher than that of Shenfu injection by 2ml/kg(P=0.0445);the MDA level of the normal resuscitation group was higher than that of the sham operation group(P<0.01),and the MDA level of the Shenfu injection 2ml/kg group was lower than normal The MDA level of the resuscitation group(P<0.01)and the Shenfu injection 7ml/kg group was lower than that of the Shenfu injection 2ml/kg(P=0.031).3.Western Blot results showed that after 24 hours of successful modeling,the normal resuscitation group compared with sham operation,the expression of cytoplasmic Nrf2 in rat myocardial tissue was significantly increased(P=0.041),and the expression of HO-1 was also significantly increased(P <0.01),indicating that the Nrf2 signaling pathway in rat myocardial tissue was activated when the model was successfully established for 24 hours,and the transcription of the downstream signaling molecule HO-1 was initiated,but P-Nrf2 was not significantly different from the sham operation group,but there was an upward trend(P =0.07),this may be related to insufficient sample size.At the same time,compared with the model group,the expression of cytoplasmic Nrf2 protein in the myocardial tissue of the Shenfu injection2ml/kg group was significantly increased(P=0.036),the expression of pNrf2 protein was increased(P=0.003),and the expression of HO-1 was significantly increased.High(P=0.001),indicating that Shenfu injection can increase the expression of Nrf2,HO-1 and p-Nrf2 protein in myocardial tissue of rats after cardiac arrest and cardiopulmonary resuscitation;Shenfu injection 7ml/kg group is better than Shenfu injection The expression of Nrf2 protein in the cytoplasm of the injection2ml/kg group increased(P=0.046),the expression of p-Nrf2 protein was significantly increased(P=0.002),and the expression of HO-1 protein was also significantly increased(P=0.009),indicating a high dose The effect of the group is better than that of the normal dose group.It shows that myocardial tissue after cardiac arrest/cardiopulmonary resuscitation activates the oxidative stress response due to ischemia-hypoxic damage,and Shenfu injection may affect the level of oxidative stress by regulating the Nrf2/HO-1 signaling pathway to protect myocardial tissue The highdose group is better than the normal-dose group.4.Real-time fluorescent quantitative PCR results showed that compared with the sham operation group,the relative expression level of HO-1m RNA in cardiac arrest rats increased(P=0.048),and the relative expression level of NQO1 m RNA increased(P=0.044);Compared with the model group,the relative expression level of HO-1m RNA in Shenfu injection 2ml/kg myocardial tissue was significantly increased(P=0.001),and the relative expression level of NQO1 m RNA increased(P=0.047);Shenfu injection 7ml/kg group Compared with the 2ml/kg group of Shenfu injection,the expression level of HO-1m RNA increased(P=0.006),and the relative expression level of NQO1 m RNA increased(P=0.041).5.The results of immunofluorescence analysis showed that the p-Nrf2 protein of cells in the sham operation group was mainly expressed in the cytoplasm;after cardiac arrest/cardiopulmonary resuscitation,p-Nrf2 was partially transferred to the nucleus;after the intervention of Shenfu injection,the p-Nrf2 protein was transferred to the nucleus The expression of p-Nrf2 protein increased significantly,and the high-dose group was better than the normal-dose group.Conclusion: The results of this study prove that Shenfu injection can effectively reduce the degree of myocardial damage in a rat model of cardiac arrest caused by asphyxia combined with ice potassium chloride.Its protective effect may be related to the anti-oxidative stress-related proteins Nrf2,HO-1,The expression of NQO1 is related. |
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