| Background:The main pathological factor,which affects the prognosis of myocardial infarction(MI),is the lost of damaged heart function,how to promote myocardial cell regeneration is the key to influence the prognosis.SIRT1 can reduce cells apoptosis under the condition of stress,which is widely known,simultaneously cardiac stem cells that have been confirmed also can be divided for good heart muscle.Traditional Chinese medicine(TCM)has an important position in the treatment of MI."Remove stasis gave birth to the new law" can regulate the expression of SIRT1 for inhibiting apoptosis and promoting stem cells to repairing damaged cardiac muscle through the early stage of Natural Science Foundation of China(NSFC),but the relevant reports of SIRT1 with cardiac stem cells is none in the treatment of MI.Objective:1.Through xuefuzhuyu capsule on rat myocardial infarction model of intervention therapy,the curative effect is observed and the internal relations will be revealed between the curative effect and the SIRT1 expression,and function of c-kit/Lin-stem cells.2.In this paper,the hypothesis of "myocardial infarction can induce cardiac stem cells proliferation and increasing SIRT1 expression can improve cardiac stem cells regeneration," remove stasis gave birth to the new law" has targeting effect of SIRT1-cardiac stem cells,so as to realizing the function of ischemic necrosis of myocardial together" will be testified.3.Revealing curative effect of therapeutic mechanism of xuefuzhuyu capsule which based on "remove stasis gave birth to the new law",provides new ideas for prevention and treatment of myocardial infarction,combined with the development of Chinese medicine in translational medicine used in regenerative medicine.Methods:1.Myocardial infarction model:The model of myocardial infarction was established by ligation of left anterior descending coronary artery.The selected rats were SPF grade healthy male WISTAR rats,200 rats weighing 230±10 g.2.Experimental group and mode of administration:a total of 200 models,survived 104,the survival rate was 52%.According to the random number table method,the rats were divided into the following eight groups:model 7 days group,Xuefuzhuyu capsule low dose 7 days group,Xuefuzhuyu capsule in the dose of 7 days group,Xuefuzhuyu capsule Dose 7 days group,model 28 days group,Xuefuzhuyu capsule low dose 28 days group,Xuefuzhuyu capsule in the dose of 28 days group,Xuefuzhuyu capsule high dose 28 days group,plus sham operation 7 days group,Sham surgery 28 days group,a total of ten groups.After the model and the second day of ECG examination,statistics I,avL,V1-V6 lead the total amount of Q wave to determine the severity of myocardial infarction,remove the degree of infarction is too heavy or too light rats,and ultimately into the observation The total number of animals was 64.Administered by intragastric administration.Sham operation group:daily with the drug group and other volume of sterile distilled water,the model group:daily with the drug group and other volume of sterile distilled water gavage,Xuefuzhuyu capsule low dose group:0.216g/kg/time,2ml/time,2 times/d,Xuefuzhuyu capsule in the dose group:0.432g/kg/time,2ml/times,2 times,Xuefuzhuyu capsule high dose group:0.864g/kg/2ml/time,2 times/d.3.ECG detection:respectively,before modeling surgery,12h after the rats were electrocardiogram detection.4.Ultrasound detection:Seven-day and 28-day experimental animals were subjected to ultrasonography using the Vevo 770 high-resolution ultrasound imaging system after 7 and 28 days of administration.Left ventricular end-diastolic margin(LVAWd),left ventricular end diastolic diameter(LVIDd),left ventricular end-diastolic diameter(LVIDs),and left ventricular short axis rate(LVPWd)(FS),left ventricular ejection fraction(EF).5.The area of myocardial infarction was measured by fluorescence irradiation with Fx Pro model.The myocardial tissue was irradiated with fluorescence.6.TUNEL method to detect myocardial cell apoptosis.7.The expression of SIRT1,FoxO1 and c-kit mRNA was detected by real-time fluorescence quantitative PCR.8.The expression of SIRT1 and Bcl-xL was detected by Western blotting.9.The expression of SIRT1 and Bcl-xL was detected by immunohistochemistry.10.ROS release was measured by reactive oxygen species after myocardial infarction.Results:1.Compared with the sham group,the left ventricular end diastolic diameter(LVIDd)and left ventricular end-diastolic diameter(LVIDs)in the model group were significantly increased at 7 days after myocardial infarction(P<0.05),and the difference was statistically significant(P<0.05)<0.01).Compared with the model group,the LVIDd and LVIDs of Xuefuzhuyu capsule low dose group and Xuefuzhuyu capsules decreased the trend of LVIDd and LVIDs,and the difference of LVIDd in Xuefuzhuyu capsule was statistically significant(P<0.01).At 28 days after myocardial infarction,LVIDd and LVIDs in model group were significantly higher than those in sham operation group(P<0.05),and the difference of LVIDs and LVIDs in Xuefuzhuyu capsule was significantly higher than that in sham operation group(P<(P<0.01).Compared with the model group,the difference of LVIDd and LVIDs between the two groups was significant(P<0.01),and compared with the dose group of Xuefuzhuyu Capsule,the low dose of Xuefuzhuyu Capsule Group LVIDs decreased significantly(P<0.01).2.Compared with the sham group,the left ventricular ejection fraction(EF)and left ventricular short axis(FS)of the model group were significantly lower than those of the sham group(P<0.01),and the difference was statistically significant(P<0.01)(P<0.05).The difference of FS in the low dose group of Xuefuzhuyu capsule and Xuefuzhuyu capsule was significantly higher than that in Xuefuzhuyu capsule(P<0.05),and there was significant difference(P<0.05),and there was significant difference between the two groups(P<0.05),and there was significant difference between the two groups(P<0.01),and there was significant difference between the two groups(P<0.01).Compared with the model group,EF and FS in the model group were significantly lower than those in the sham operation group(P<0.01)(P<0.01).Compared with the middle dose group of Xuefuzhuyu Capsule,Xuefuzhuyu Capsule and Xuefuzhuyu Capsule were significantly higher than those in Xuefuzhuyu Capsule There was significant difference in FS group between low dose group(P<0.05).3.Compared with the sham operation group,the myocardial infarct size in the model group increased significantly,and the difference was statistically significant(P<0.01).Compared with the model group,the myocardial infarct size of the model group was significantly higher than that of the model group(P<0.01).Compared with the model group,the myocardial infarct size in the model group was significantly lower than that in the control group(P<0.01),and the myocardial infarct size in the model group was significantly lower than that in the sham operation group(P<0.05),and there was significant difference between the two groups(P<0.05).4.The results of TUNEL showed that the apoptotic rate of the model group was significantly higher than that of the sham operation group(P<0.01),compared with the sham operation group(P<0.01).Compared with the model group,the apoptotic rate of the model group was significantly higher than that of the model group(P<0.01).Compared with the model group,the difference was statistically significant(P<0.01).The apoptotic rate was significantly decreased and the difference was statistically significant(P<0.05).5.The expression of SIRT1 and C-kit in model group was significantly lower than that in sham operation group(P<0.01)(P<0.05),and the expression of FoxO1 was significantly increased(P<0.01)(P<0.05).The expression of SIRT1 and C-kit increased significantly(P<0.05,P<0.01),FoxOl expression was significantly decreased(P<0.01);28 days after myocardial infarction,compared with the model group,the expression of SIRT1 and C-kit in Xuefuzhuyu capsule group was significantly increased(P<0.01),and the expression of Foxin was significantly increased(P<0.01)(P<0.01),FoxOl expression was significantly decreased(P<0.01).6.The expression of Bax and Bcl-xL protein in myocardium was significantly higher than that in sham operation group(P<0.05).The expression of Bax and Bcl-xL protein in model group was significantly higher than that in sham operation group(P<0.05)(P<0.05).Compared with the model group,the expression of Bax protein in Xuefuzhuyu capsule group was significantly lower than that in the model group(P<0.05),the expression of Bcl-xL protein was significantly increased and the difference was statistically significant(P<0.01).Compared with the model group,the expression of Bax protein in the model group was significantly higher than that in the sham operation group(P<0.01),and the expression of Bcl-xL protein in the model group was significantly higher than that in the model group The expression of Bax protein was significantly decreased and the expression of Bcl-xL protein was significantly increased and the difference was statistically significant(P<0.05).7.The expression of Caspase-3 and Fas protein in the model group showed that the expression of Caspase-3 and Fas protein in the model group was significantly higher than that in the sham operation group at 7 days after myocardial infarction and the difference was statistically significant(P<0.05).Compared with the model group,the expression of Caspase-3 protein in Xuefuzhuyu capsule group was significantly lower than that in model group(P<0.05),and the difference was statistically significant(P<0.05)(P<0.01).The expression of Caspase-3 and Fas protein in model group was significantly higher than that in sham operation group at 28 days after myocardial infarction(P<0.01),and the expression of Fas protein was significantly lower than that in sham operation group(P<0.01),The expression of Caspase-3 and Fas protein in Xuefuzhuyu capsule group was significantly lower than that in model group(P<0.01).8.The expression of SIRT1 and SOD2 protein in the model group showed that the expression of SIRT1 and SOD2 protein in the model group was significantly lower than that in the sham operation group(P<0.01),and the expression of SIRT1 and SOD2 protein in the model group was significantly lower than that in the sham operation group(P<0.05).The expression of SIRT1 and SOD2 protein in the model group was significantly lower than that in the sham operation group(P<0.05).The expression of SIRT1 and SOD2 protein in the model group was significantly lower than that in the sham operation group(P<0.05)(P<0.01).Compared with the model group,the expression of SIRT1 and SOD2 protein in Xuefuzhuyu capsule group was significantly increased(P<0.01).9.The expression of FoxOl and P53 protein in myocardium was significantly higher than that in sham operation group(P<0.01),and the expression of P5 3 protein was significantly increased(P<0.01)(P<0.01).The expression of P53 protein was significantly decreased(P<0.01).Compared with the model group,the expression of FoxOl protein in the model group was significantly lower than that in the model group(P<0.01)(P<0.01).Compared with the model group,the expression of FoxOl and P53 protein in Xuefuzhuyu capsule group was significantly decreased and the difference of FoxOl protein expression was significantly statistically significant(P<0.01),but there was no significant difference between the two groups(P<0.01).The difference of P53 protein expression was statistically significant(P<0.05).10.Immunohistochemistry The color intensity of positive cells was measured by measuring the density of the cells and the relative optical density was calculated.The results showed that SIRT1 and Bcl-xL staining at 7 days and 28 days at each time point(P<0.05).The intensity of SIRT1 and Bcl-xL in Xuefuzhuyu capsule group was significantly higher than that in model group(P<0.05),and the intensity of Fas and P53 was significantly increased(P<0.05).The staining intensity of Fas and P53 was significantly lower than that of model group(P<0.05).(P<0.01).11.Compared with the sham operation group,the ROS of the model group was significantly higher than that of the model group(P<0.01).Compared with the model group,the ROS of the Xuefuzhuyu capsule group was significantly higher than that in the sham operation group at 7 days and 28 days after myocardial infarction Significantly decreased(P<0.05).Conclusion:1.Xuefuzhuyu capsule by improving ventricular remodeling,improve heart function,and thus play its role in cardioprotective effect.2.Xuefuzhuyu capsule through anti-cardiomyocyte apoptosis,reduce myocardial infarct size,and thus play its role in cardiac regeneration.3.Xuefuzhuyu capsule has improved ventricular remodeling,anti-cardiomyocyte apoptosis,the mechanism may be related to its inhibition of ROS production.4.Xuefuzhuyu capsule with anti-cardiomyocyte apoptosis,and promote the role of cardiac stem cell regeneration.5.Increased expression of SIRT1 can improve the regeneration of cardiac stem cells.Xuefuzhuyu capsule may have the effect of targeting SIRT1-cardiac stem cells to achieve the common repair of ischemic myocardium.6.SIRT1 pathway may be a potential target for maintaining cardiac stem cell function. |
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