Yiqi Huoxue Traditional Chinese Medicine For Chronic Heart Failure Rats Myocardial Effect Of Mmp 1 And Timp - 1 Experimental Study

Purpose：Based on the model of chronic heart failure (CHF) rats died of myocardialinfarction, this thesis observes reaction and leaf heart effect of MMP-1andTIMP-1on CHF rats by Qi and activating Blood compound prescription, and adoptsRT-PCR, Western blotting, immunohistochemistry to investigate mechanism ofaction of CHF by Qi and activating Blood compound prescription.Material and method：1. Model of chronic heart failure (CHF) rats establishing: select60weighted250-300g, aged12ordinary rats, feed one week, and select randomly10rats ascontrol group, and the rest establishing model of chronic heart failure (CHF)rats by coronary artery ligation(CAL) in conjunction with Exhaustive Swimming,food lowering, then test left venticular ejection fraction(LVEF)≤60%byDoppler echocardiogram, and find rats psy-chasthenic and their fur not shiny,which indicates model establishment done.2. Rats grouping:8rats died during the modeling process. successful modelof rats (24rats)were randomly divided into chronic heart failure model group(8rats),benifiting qi and actvating bloos compound recipe group(8rats), westernmedicine group(8rats), sham operation group(8rats)is control group.3. Medicine feeding: we feed by Qi and activating Blood compound prescriptioncompounded by astragalus, ginseng, safflower, salvia, leonurus japonicus,panax notoginseng powder and semen lepidii. Rats in medication group fed9.2gmedicines/Kg/d or1.5mg lisinopril/Kg/d medicine group by mice and mendose formula.6weeks later, all rats were killed after anesthesia, left theheart myocardium. Control group and model group are only fed distilled waterof conventional volume for6weeks. 4. Heart function check: after6weeks of distilled water feeding, checkthe rats by Doppler echocardiogram, test their left ventricular internaldiameter at end-systole（LVIDs）, Left Ventricular Internal Dimension-Diastole(LVIDd), and Left Ventricular Ejection Fraction (EF%).5. By the real time quantitative PCR, gene expression of MMP-1, TIMP-1inmyocardial tissue at each group were measured.6. Using Western blot method, protein expression of MMP-1, TIMP-1inmyocardial tissue at each group were measured.7. HE staining, and observing morphological changes in myocardial tissueunder light microscope among groups.8. Observing morphological changes in MMP-1、TIMP-1in myocardial tissueby immunohistochemical assay(IHCA).Results：1. Chronic heart failure animal model group LVIDs、LVIDd was significantlyincreased compared with the control group; by drugs intervention, LVIDs、LVIDdof each treatment group was significantly decreased, and EF was increasedobviously.(1)Regarding to LVIDs, it was significantly decreased among Chinesemedicine group and Western group, compared with model group（P＜0.01）; whilethey compare each other, P＞0.05.(2)Regarding to LVIDd, it was significantlydecreased among Chinese medicine group and Western group, compared with modelgroup（P＜0.01）; while they compare each other, P＜0.05.(3)Regarding to EF%,it was significantly increased among Chinese medicine group and Western group,compared with model group（P＜0.01）; while they compare each other, P＞0.05.2. Chronic heart failure animal model group MMP-1mRNA expression wassignificantly increased compared with the control group(P＜0.01）; by drugsintervention, MMP-1mRNA of each treatment group was significantly decreasedcompared with model group(P＜0.01）.Chinese medicine group was decreasedobviously,53%of model group(P＜0.01）; and Western group is0.58%of modelgroup. Chinese medicine group vs Western group,statistically significant(P＜ 0.05）3. Chronic heart failure animal model group MMP-1protein expression wassignificantly increased compared with the control group(P＜0.01）; by drugsintervention, MMP-1protein expression of each treatment group wassignificantly decreased compared with model group(P＜0.01）.Western group wasdecreased obviously, and it is46%of model group; and Chinese medicine groupis half of model group. Chinese medicine group vs Western group, statisticallysignificant(P＜0.05）4. Chronic heart failure model group Timp-1protein expression wassignificantly reduced compared with the control group (P <0.01); by drugintervention the Timp-1mRNA of each treatment group was significantly increasedcompared with model group(P＜0.01）,and Chinese medicine group increasedsignificantly, that is2.5times model group. Western group is2.05times modelgroup. Western group vs Chinese medicine group, statistically significant(P＜0.05）5. Chronic heart failure model group Timp-1protein expression wassignificantly reduced compared with the control group (P <0.01); by drugintervention the Timp-1mRNA of each treatment group was significantly increasedcompared with model group(P＜0.01）,and Chinese medicine group increasedsignificantly, that is2.09times model group. Western group is1.93times modelgroup. Western group vs Chinese medicine group, statistically significant(P＜0.05）6. HE Histological observation showed that in the blank control groupthe myocardial cells were regular arrangement, stripes clear, myocardial nucleilocated in the myocardial cell edge, its shape and size were normal.Orderlyarrangement of myofibrils still shows that there were not faults,defects andthe blank area. Model group: the myocardial cells were intertwined and had nostripes and out of shape. The myofibril were deranged, broken and lossed. Themyocardial nuclear were pyknosis.There were some white space and theheterochromatin were lumpish. Adjacent cells intercalated disk space widened and the tight junctions connecting capsules disappeared. Morphology ofmyocardial cells in all treatment groups compared with the model group wereimproved, Western group improved most significantly.7. Using immunohistochemistry，the average gray of Chronic heart failuremodel group MMP-1was550.14±132.83compared with the control group，therewas statistically significant(P <0.01); Western group impression significantlywas114.72±46.26compared with model group, statistically significant(P <0.01);Chinese medicine group was124.98±30.40compared with model group, there wasstatistically significant (P <0.01). There was no significant differenceamong Chinese medicine and Western group（P＞0.05）.8. Using immunohistochemistry,the average gray of Chronic heart failuremodel group Timp-1was115.83±43.25compared with the control group, there wasstatistically significant(P <0.01); Western group was181.16±55.34comparedwith model group, there was statistically significant(P <0.01); Chinesemedicine group was172.94±48.75compared with model group, there wasstatistically significant(P <0.01); There is no significant difference amongChinese medicine and Western group（P＞0.05）.Conclusion：1. Qi and activating Blood compound prescription by6weeks after treatment,can improve the level of LVIDs, LVIDd,and increased the level of EFin chronicheart failure rats, which will protect the heart function and inhibit the heartchamber to expand.2. Using real-time fluorescence quantitative PCR, Western blot,immunohistochemical detection of rat myocardial tissue, found that Qi andactivating Blood compound prescription inhibit MMP-1protein and gene expression,increase TIMP-1protein and gene expression, reduce MMP-1/TIMP-1ratio, preventMMP-1on myocardial collagen degradation，coordinate degradation and synthesisof ECM，inhibit or reverse ventricular remodeling, improve myocardial compliance,increases myocardial contractility, and playe the role of treatment of chronic heart failure.3. Increase of MMP-1protein and gene expression, decrease of TIMP-1proteinand gene expression, imbalance of MMP-1/TIMP-1ratio, participate in the processof left ventricular remodeling.4. Qi and activating Blood compound prescription has the effect of Qi andblood circulation, benefits of water swelling. It can improve heart function,inhibite ventricular remodeling, which is better than western medicine group.It provides scientific basis and effective way for the rehabilitation of chronicheart failure.