| Objective To observe the effect and mechanism of Yiqi Yangyin and Wenyang Lishui treatment of chronic heart failure.methods 1.The establishment of the model: The rat model of chronic heart failure induced by pressure overload was established by abdominal aortic coarctation(AAC).The sham operation group received the same operation but not abdominal aortic coarctation.2.Grouping and administration: 6 weeks after the operation,the survival model rats were randomly divided into three groups: model group,Qili group,Yiqi Yangyin and Wenyang Lishui Prescription group(traditional Chinese medicine group),10 rats in each group;the remaining 10 rats in the sham operated group rats as normal control.Qili group was administrated Qili Qiangxin capsule into powder suspension with 0.1g/100 g dose timing by gavage once a day;the traditional Chinese medicine group at a dose of 12.13g/kg orally once timing;model group and sham operated group were given saline gavage.4ml one time;drug intervention for 4 weeks.3.Determination of the respiratory frequency: Before modeling establishment,10 rats were randomly selected from 62 healthy SD rats to test frequency respiratory when they were in silence(at about 8:00-10:00 in the morning),calculating the average value;before drug intervention and ater 4 weeks’ drug intervention,frequency respiratory of each rat were also tested and recorded.After 6 weeks of modeling in drug intervention and drug intervention of each rat respiratory frequency and record the mean calculated after 4 weeks comparison;in rats at different time points of the respiratory frequency change.4.Collection of samples: after 4 weeks of continuous drug intervention, weighed rats and recorded BW.Rats were anesthetized by intraperitoneal injection of 10% chloral hydrateat at a dose of 0.3ml/100 g,and all rats were detected by color Doppler echocardiography to assess the changes of cardiac structure and function.The blood samples were collected from the abdominal aorta,and the serum concentrations of BNP,MMP-2 and Ang II were detected by ELISA.After bled to death,their hearts and left ventricular tissues were separated immediately,rinsing with ice saline and drying with filter paper.After weighing,recording heart mass(HM),left ventricular mass(LVM),calculated the cardiac mass index(HMI)and left ventricular mass index(LVMI),HMI=HM/BW,LVWI=LVM/BW.Ventricular tissue was divided into two parts.One part was fixed in the 4% Polyoxymethylene solution and saved in 4℃ refrigerator for HE staining and Massion staining for the purpose of myocardial cell morphology observation and detection of myocardial fibrosis and myocardial collagen volume fraction calculation;the other part was put into the refrigerator at-80℃ for testing the expression level of myocardial TGF-β 1 mi R-21 and SPRY2.Protein content of myocardial TGF-β 1 was determined by Western blot,and the expression of mi R-21 and SPRY2 m RNA by rt-q PCR.5.Results: 10 weeks after AAC,compared with sham group,rats accepted AAC operation had decreased activity decreased,reduced intake of diet and water,the respiration rate,especially in the model group.Rats in traditional Chinese medicine group and Qili group were better in general state compared with model group.The indicators of cardiac hypertrophy showed that the HMI and LVMI in the model group were significantly higher than that of the sham operation group(P < 0.01);Echocardiography showed that the level of indicators including LVEDD,LVESD,LVESV,LVEDV,LVPWI,IVST,E/e’in model group were significantly higher than that in the sham group(P < 0.05 or P < 0.01);the 9 indicators above of traditional Chinese medicine group and Qili group were higher than that of the sham operation group,lower than that of model group(P < 0.05);Chinese medicine group decreased HWI and LVWI compared with Qili group(P < 0.01);there was no significant difference between the Chinese medicine group and Qili group in LVEDD,LVESD,LVEDV,LVESV,LVPWT,IVST,E/e(P > 0.05);lower LVEF and FE was detected in the model group than in the sham operation group(P < 0.05);higher LVEF and FE was detected in Qili group and Chinese medicine group than in model group(P < 0.05).However,the LVEF and EF of the four groups were more than 50%.HE staining showed no hypertrophy 、 interstitial hyperplasia andinflammatory cell infiltration with normal cell morphology in the myocardial cells of rats in sham operation group.Rats in model group showed obvious myocardial cell disorder,cell hypertrophy,interstitial hyperplasia,inflammatory cell infiltration.The Chinese medicine group and Qi Li group showed that myocardial cells arranged regularly and displayed slight hypertrophy and minimal stromal hyperplasia as well as slight inflammatory cell infiltration.The two groups showed no significant visible difference.Masson staining showed that the model group compared with the sham operation group showed obvious myocardial fibrosis and increased collagen volume fraction(P < 0.01);traditional Chinese medicine group and Qili group showed reduced myocardial fibrosis compared with the model group,and the collagen volume fraction decreased(P < 0.01);there were no significant difference in collagen volume fraction between traditional Chinese medicine group and model group(P > 0.05).Serological tests showed that BNP,MMP-2,Ang II serum concentration of the model group was significantly higher than that of sham operation group(P < 0.05);Chinese medicine group and Qili group showed higher level of BNP,MMP-2,AngⅡ than the sham operation group did(P < 0.05),and lower level than the model group(P < 0.01).Western blot showed that expression level of TGF-β 1 in the model group,Chinese medicine group and Qili group were higher than those in sham operation group(P < 0.01);Chinese medicine group and Qili group decreased TGF-β 1 expression compared with the model group(P < 0.05);there was no significant difference between the traditional Chinese medicine group and Qili group(P > 0.05).rt-q PCR showed that the m RNA level of mi R-21 in model group was increased significantly compared with the sham operation group(P < 0.01);the level of mi R-21 in Qili group and Chinese medicine group was lower than the model group(P < 0.05),higher than the sham operation group(P < 0.01).the m RNA level of SPRY2 in Chinese medicine group,model group and Qili group was lower than that in sham operation group(P < 0.05);Qili group and Chinese medicine group elevated the expression of SPRY2 compared with control group(P < 0.05);Qili group and Chinese medicine group showed no significant difference in mi R-21 and SPRY2 m RNA expression(P > 0.05).Conclusion 1.The hypertrophy of myocardium and myocardial fibrosis in chronic heart failure rats were related to the increase of serum BNP,MMP-2,Ang II and the protein overexpression of TGF-β 1 in myocardium.2.Yiqi yangyin and Wenyang Lishui treatment can reduce the cardiac hypertrophy and myocardial fibrosis,resulting in inhibiting ventricular remodeling and improving cardiac function.3.Yiqi yangyin and Wenyang lishui formula can decrease the expression of serum Ang,BNP,MMP-2 and the level of myocardial TGF-β 1 to reverse ventricular remodeling and improve cardiac function.4.Yiqi yangyin and Wenyang lishui treatment has a cardiacprotective effect on chronic heart failure rats by depressing m RNA expression of mi R-21 and upregulating SPRY2 m RNA expression,which importantly involved in activation of MAPK signaling pathway and EMT process. |
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