| PurposeThis experiment study for Yiqihuoxue compound treatment of chronic heartfailure after myocardial infarction in rats. Observe AT1receptor and ERK2changes before and after treatment during the process of ventricular remodelingin chronic heart failure. Explore the mechanism and effect of yiqi huoxuecompound reverse left ventricular remodeling in heart failure. Provide ascientific and reliable experimental research method for the treatment oftraditional Chinese medicine compound in chronic heart failure.Material and method1. Establish chronic heart failure model of blood stasis due to qi deficiency:Choose one hundred regular grade12weeks old healthy male wistar rats, whichbody weight between180g to220g. Through the operation of chest coronary arteryligation. Dynamic display of chest lead ST segment arch upward inelectrocardiogram. With method of exhaustion swimming type and underfeed sixweeks after the operation, through heart Doppler calculate left ventricularejection fraction≤40％or cardiac index≤180ml/min/kg as a symbol of buildingchronic heart failure model successful. Those chronic heart failure model hadmanifestation of weight continued to drop, dispirited, ecchymosis in claw,edema, which belong to blood stasis due to qi deficiency type in traditionalChinese medicine.2. Experimental animal groups: Divided survival model rats into four groups,normal group, control group, Chinese medicines group and western medicine group.Each group includes10rats.3. Experimental drugs: Yiqihuoxue compound contains milkvetch root,motherwort herb, safflower, danshen root, sanqi, Indian bread, dried freshginseng, pepperweed seed, which provided by Jiangyin Tianjiang Pharmaceutical.Western medicine group use Lisinopril which provided by Shantou Jinshipharm, each pill dose5mg.4. Drug dose: Normal group and control group rats without drug intervention,quantitative volume of distilled water to fill the stomach everyday. Treatmentgroup rats dose on the basis of conversion formula，Chinese medicines group oncedose4.6g Yiqihuoxue compound per kilogram, western medicine group once dose1.5mg Lisinopril per kilogram. Once per day, continuous lavage fed for6weeks.5. Ultrasonic Doppler examination of cardiac function: After6weeks with drugor distilled water to fill the stomach, continuous detecting left ventricularend systolic diameter(mm) and left ventricular end diastolic diameter(mm)inthree different cardiac cycle, taking average value, calculating leftventricular ejection fraction and fractional shortening.6. The ratio of rat heart weight/body weight: After groups of rats have finishedcardiac Doppler examination, put rats to death immediately. Get rat heart weightand body weight, calculating the ratio of rat heart weight/body weigh, whichis heart weight index.7. Through haematoxylin and eosin stain, observe groups of rats myocardialmorphology changes under microscopic.8. Measure each group rats’ myocardial tissue AT1-mRNA, ERK2-mRNA expressionlevel by real-time fluorescent quantitative PCR technique.9. Observe each group rats’ myocardial tissue AT1, ERK2changes byimmunohistochemistry, and using Image－Pro Plus to measure average gray value.10. Use statistics software to analyze each group rats’ myocardial tissueAT1-mRNA, ERK2-mRNA and protein expression levels and those that therelationship between the ratio of heart weight/body weigh and left ventricularejection fraction, fractional shortening.Results1. Ultrasonic Doppler examination of cardiac function，control group rats’left ventricular ejection fraction and fractional shortening reducedsignificantly, and compared with normal group rats, P<0.01，date difference are significantly. After treatment with drugs, treatment groups rats’ leftventricular ejection fraction and fractional shortening raised significantly,and compared with control group rats, P<0.05，date difference are significantly.Compared between Chinese medicine and western medicine group, P<0.05, datedifference are significantly. These prompt that Yiqihuoxue compound is superiorto western medicine lisinopril in the aspect of improving left ventricularejection fraction and fractional shortening.2. Compared with normal group rats, control group rats’ cardiac weight indexraised significantly, P<0.01，date difference are significantly. After treatmentwith drugs, treatment groups rats’ cardiac weight index reduced significantly,and compared with control group rats, P<0.05，date difference are significantly.Compared between Chinese medicine and western medicine group, P>0.05, datedifference are not significantly. These prompt that Yiqihuoxue compound is equalto western medicine lisinopril in the aspect of improving rats’ cardiac weightindex.3. Then watch HE staining groups in heart failure rat myocardial tissuemorphology changes: Normal groups rats’ myocardial cells is neatly, that rundisk, band is clear, morphology, structure is conformity, nucleus is locatedin the edge and myofibril arranged orderly. Control group rats’ myocardial cellinterwoven into a network, that can’t see the horizontal stripes, karyon, musclefibers arranged disordered, even fractured and appearing a blank area. Westernmedicine group rats’ myocardial cell arrangement is neatly, that the morphology,size of nucleus is basic normal, myofibril arranged orderly, but the defectsand blank area are still visible. Chinese medicine group rats’ myocardial cellarrangement is neatly, that nucleus is located in the edge, myofibril arrangedorderly, but the defects and blank area are still visible.4. Compared with normal group rats, the expression of AT1-mRNA, ERK2-mRNAin control group rats’ myocardial cell increased significantly, P<0.01，thatdate difference are significantly. Compared with control group, each treatmentgroup after drug intervention AT1-mRNA, ERK2-mRNA decreased obviously, P<0.05, that date difference are significantly. Compared between Chinese medicine andwestern medicine group, P>0.05, date difference are not significantly. Theseprompt that Yiqihuoxue compound is equal to western medicine lisinopril in theaspect of pulling down the expression of AT1-mRNA, ERK2-mRNA5. Using immunohistochemical method analysis the different expression of AT1,ERK2protein in each group rats’ myocardial tissue, and using Image－Pro Plusto measure average gray value. The expression of AT1, ERK2protein in controlgroup rats’ myocardial cell increased significantly, P<0.01，that datedifference are significantly. After treatment with drugs, the expression ofAT1,ERK2protein in treatment groups rats’ myocardial cell reducedsignificantly, and compared with control group rats, P<0.05，that datedifference are significantly. Compared between Chinese medicine and westernmedicine group, P>0.05, date difference are not significantly. These prompt thatYiqihuoxue compound is equal to western medicine lisinopril in the aspect ofpulling down the expression of AT1ERK2protein.6. The expression of AT1, ERK2gene and protein in chronic heart failure rats’myocardial tissue has a relationship with the ratio of rat heart weight/bodyweight, left ventricular ejection fraction and fractional shortening, P<0.01，that date difference are significantly.Conclusion1.By observing the behavior of rats and morphological changes in rats’myocardial tissue, texting heart Doppler, calculating left ventricular ejectionfraction, it proved that a blood stasis due to qi deficiency animal model ofchronic heart failure can be successfully established though ligature rat’coronary artery with exhaustion swimming and eating less.2. Yiqi huoxue compound could improve heart failure rats’ left ventricularejection fraction and fractional shortening, reduce the ratio of rat heartweight/body weight, delay or reverse left ventricular remodeling in heartfailure. 3. Left ventricular remodeling in rats with chronic heart failure cardiacmuscle tissue in the rise of AT1, ERK2.4. Yiqi huoxue compound could reduce the expression of AT1, ERK2in chronicheart failure rats’ cardiac muscle tissue, relieve the symptoms, improvecardiac function, delay or reverse myocardial tissue structure which was alreadyhappened refactor.5. Yiqihuoxue compound is equal to western medicine lisinopril in the aspectof improving cardiac function, delaying or reversing left ventricular remodelingin heart failure. Provide a scientific and reliable experimental research methodfor the treatment of traditional Chinese medicine compound in chronic heartfailure. |
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