Experimental Study Of Shenzhuxinkang Decoction Regulating MiRNA-21 In Anti-cardiac Fibrosis End-MT In Rats With Heart Failure

Objective: To explore the effects of Shenzhu Xinkang Decoction on mi RNA-21,PTEN/PI3K/Akt signaling pathway and End-MT,and to explore the internal molecular mechanism of Shenzhu Xinkang Decoction in improving myocardial fibrosis in chronic heart failure.The application provides experimental basis.Methods: Ten of 55 SD rats were randomly selected as the sham operation group.The remaining rats used the abdominal aorta constriction method to replicate the chronic heart failure rat model.The sham operation group only used sutures to pass through the abdominal aorta without ligation.The rest of the operation steps are the same as the model building.The general condition of the rats was observed and recorded after the operation.Eight weeks later,3 rats were randomly selected from the sham operation group and the model group for echocardiography to prove the successful replication of the model.After confirming that the model was successfully copied,the rats in the model were randomly divided into model group,captopril group,and Shenzhu Xinkang decoction group.Both the sham operation group and the model group were intragastrically administered with distilled water(10ml/kg).Captopril(7.8 mg/kg)was given to the Prili group,and the Shenzhu Xinkang Decoction(44g/kg)was given to the Shenzhu Xinkang Decoction group.The stomach was given once a day for 6 weeks.After 6 weeks,Masson staining and HE staining were used to detect myocardial fibrosis in rats,Elisa method was used to detect myocardial Hyp content,cellular immunofluorescence staining was used to detect myocardial CD31,?-SMA,and FSP1 protein expression levels,and RT-PCR was used to detect mi RNA.-21,PTEN,PI3 K,Akt m RNA expression level,Westernblot detects the protein expression level of PTEN,PI3 K,Akt,p-Akt.Results:1.General conditions of rats and echocardiographic examinationRats in the sham-operated group showed no obvious abnormalities.The model rats all showed typical heart failure manifestations such as decreased activity,slow response,shortness of breath,irritability,limb edema,and decreased urine output.Eight weeks later,3 rats were randomly selected from the sham operation group and the model group for echocardiography.Compared with the sham operation group,the model group had an increase in LVIDd(P<0.05),and both LVIDs and LVESV Significant increase(P<0.01),and EF and FS values decreased significantly(P<0.01),proving the success of model building in rats with heart failure.After group intervention for 6 weeks,the heart failure performance of rats in the captopril group and the Shenzhu Xinkang decoction group was significantly improved compared with the model group.2.Results of myocardial pathological staining and myocardial hydroxyproline(Hyp)content in ratsMasson staining and HE staining showed that the myocardial fibrosis in the model group was obvious,and after 6 weeks of administration of captopril and Shenzhu Xinkang decoction,the myocardial fibrosis of rats was improved.Detection of myocardial Hyp content: the hydroxyproline content of myocardial tissue in the model group was significantly increased(P<0.01);compared with the model group,the Hyp content of myocardial tissue in the captopril group was decreased(P < 0.05),the Shenzhuxinkang decoction group The Hyp content of myocardial tissue was significantly reduced(P<0.01);compared with the captopril group,the Hyp content of myocardial tissue was reduced in the Shenzhuxinkangtang group,and the difference was not statistically significant(P>0.05).3.Cellular immunofluorescence staining resultsA large number of cells in the sham operation group are positive for CD31,and only a small number of cells are positive for ?-SMA;a large number of cells in the model group are positive for ?-SMA,only a few cells are positive for CD31,and the expression of CD31 in the model group Compared with the sham operation group,the expression of ?-SMA was significantly increased(P<0.01);a large number of cells in the Shenzhu Xinkang decoction group and the captopril group expressed CD31 positive,and CD31 The expression of ?-SMA was significantly higher than that of the model group(P<0.01),only a few cells were positive for ?-SMA,and the expression of ?-SMA was lower than that of the model group(P<0.05).Compared with the sham operation group,a large number of cells in the model group expressed FSP1 positive(P<0.05).Compared with the model group,only a few cells in the Shenzhuxinkangtang group and captopril group expressed FSP1 positive(P<0.05).There was no significant difference between the test results of the captopril group and the Shenzhu Xinkang decoction group(P>0.05).4.m RNA expression of mi RNA-21,PTEN,Akt,PI3 K in each groupCompared with the sham operation group,the m RNA expression of mi RNA-21,Akt and PI3 K in the model group,captopril group,and Shenzhu Xinkang decoction group increased significantly(P<0.05),and the expression of PTEN m RNA decreased significantly(P<0.01);Compared with the model group,the m RNA expression of mi RNA-21,Akt and PI3 K in the captopril group and the Shenzhu Xinkang decoction group was significantly decreased(P<0.01),and the expression of PTEN m RNA was significantly increased(P < 0.01),compared with the captopril group,the m RNA expression of mi RNA-21 and Akt in the Shenzhu Xinkang Decoction group was significantly decreased(P<0.01),the expression of PTEN m RNA was significantly increased(P<0.01),and the m RNA expression of PI3 K Decrease(P<0.05).5.Protein expression of PTEN,Akt,PI3 K,and p-Akt in each groupCompared with the sham operation group,the expression of PTEN protein in the model group,captopril group,and Shenzhu Xinkang decoction group was significantly reduced(P<0.01).PI3 K,the model group,the captopril group,and the Shenzhu Xinkang decoction group were significantly reduced.The expression of Akt and p-Akt protein increased significantly(P<0.01).Compared with the model group,the expression of PTEN protein in the captopril group and the Shenzhu Xinkang decoction group was significantly increased(P<0.01);the PI3 K,Akt,and p-Akt proteins in the Shenzhu Xinkang decoction group and the captopril group The expression decreased significantly(P<0.01);compared with the captopril group,the PTEN protein expression in the Shenzhu Xinkang decoction group was significantly increased(P<0.01),and the Akt and p-Akt protein expressions were significantly decreased(P<0.01).The expression of PI3 K decreased(P<0.05).Conclusion:Shenzhu Xinkang Decoction can inhibit the proliferation of myocardial collagen fibers,significantly improve myocardial fibrosis in CHF rats,and can effectively treat CHF.Shenzhu Xinkang Decoction can down-regulate the m RNA and protein expression of mi RNA-21,PI3 K,Akt,up-regulate the expression of PTEN m RNA and protein,and inhibit End-MT.The effect of Shenzhu Xinkang Decoction against myocardial fibrosis may be related to its ability to regulate the genes and proteins of the PTEN/PI3K/Akt pathway by inhibiting the expression of mi RNA-21.Shenzhu Xinkang Decoction can inhibit myocardial fibrosis by inhibiting End-MT,and the inhibition of End-MT by Shenzhu Xinkang Decoction may be related to the regulation of mi RNA-21.