| Purpose:The experiment was to explore the relation between the matrix metalloproteinase (MMP-1), matrix metalloproteinase tissue inhibitor (TIMP-1) , MMP-1 / TIMP-1 and left ventricular remodeling of chronic heart failure and the mechanism of benifiting qi and actvating bloos compound recipe allying exercise training profecting cardiac function and reducing left ventricular remodeling in chronic heart failure rats by establishing the chronic heart failure modol ,benifiting qi and actvating bloos compound recipe allying exercise training as an intervention factor.Then providing the scientific basis and effective way of the mechanism of chronic heart failure and rehabilitation treatment.Material and method:1. Creating the the model of chronic heart failure in rats: Take the ordinary level 60 healthy SD male rats, weighing 200 to 230 grams, and ligated coronary artery with exhaustive style swimming and hunger mehtods to create after 1 week feeding adaptation.Then to determine wether the the model of chronic heart failure in rats was successful or not by detecting left ventricular ejection fraction (EF)≤60% in Doppler echocardiography, and observing the state of the rats, cardiac muscular tissue morphology.2. Grouping the experimental animal: Eight rats died during the modeling process. successful model of rats (44 rats)were randomly divided into chronic heart failure model group(8 rats) ,benifiting qi and actvating bloos compound recipe group(8 rats), exercise training group(10 rats), benifiting qi and actvating bloos compound recipe allying exercise training group(10 rats), western medicine group(8 rats), sham operation group(8 rats)is control group. 3.Selecting medicine: benifiting qi and actvating bloos compound recipe consists of astragalus, ginseng, safflower, red sage root, motherwort, Pseudo-ginseng powder, semen lepidii etc. Granular preparation of benifiting qi and actvating bloos compound recipe is provided by the third pharmaceuticat factory in Benxi. Lisinopril tablets of western medicine group are produced by Shanghai Xinyi Pharmaceutical Company, each piece 10Mg.4.Giving medicine and doing exercise training to experimental animals: in addition to heart failure model group and sham operation group, other groups were given drug treatment and(or) exercise training after successful modeling for 6 weeks. Control group and model group were treated with conventional equal volume of saline, given medicine groups were administered dose at conversion formula of human and mice.Western medicine was given Lisinopril at 1.5mg crude drug / Kg / d, Chinese medicine group was given benifiting qi and actvating bloos drug at 9.2g crude drug / Kg / d. Exercise training was done after 1 week in the successful modeling. According to the official physical therapy using rat body weight / oxygen consumption regression equation, ET model[1].Intermittent training, each exercise 15 min, rest 5 min.Treadmill exercise training conducted at an initial treadmill angle of 5°, a speed of 10 m / min, and had a rest 10 minutes. Increasing training intensity, and ultimately achieved at a speed of 16 m / min, 50 min exercise break (including 5.minutes warm-up training), and in the course of the experiment to maintain this amount of exercise,5 days a week. Rats were treadmill for 5 min in order to adapt to treadmill running, and then adjusted to 5 degrees, 10 m / min (equivalent to the maximum oxygen uptake 45% VO2max).First time ran 15 min,second time 30 min,third time 45 min and in the course of the experiment to maintain this amount of exercise, 5 days a week, training for 5 weeks.5. Detecting the cardiac function by echocardiography: before and after medicine treatment and (or) exercise training, each group was made on echocardiography, To measure the left ventricular systolic and diastolic final internal diameter (LVIDs and LVIDd, mm) in three different continuous cycle by M-mode echocardiography, and took the the mean value to calculate the left ventricular ejection fraction, left ventricular end systolic and diastolic volume, output and per Bo fractional shortening.6.By routine HE staining, the rats were observed morphological changes of myocardial cells.7.By the real time quantitative PCR, gene expression of MMP-1, TIMP-1, MMP-1 / TIMP-1 ,Ⅰcollagen,Ⅲcollagen in myocardial tissue at each group were measured.8.By Western-blot, the protein levels of MMP-1, TIMP-1, MMP-1 / TIMP-1 ,Ⅰcollagen,Ⅲcollagen in myocardial tissue at each group in myocardial tissue of each group were determined.9.By the immunohistochemical method, the morphological changes of MMP-1, TIMP-1, MMP-1 / TIMP-1 ,Ⅰcollagen,Ⅲcollagen in myocardial tissue at each group were detected .Results:1. In model group the value of EF,FS and SV decreased significantly and LVEDV, LVESV increased significantly compared with sham control group(P<0.01). After medicine treatment and (or) exercise training, the value of EF, FS and SV at treatment groups were increased, and the value of LVEDV and LVESV were decreased significantly compared with the model group (P<0.01). (1) For the EF, traditional Chinese medicine combined exercise group increased most significantly and had statistical significance compared with the Chinese medicine group, western medicine group and exercise group (P<0.01). Traditional Chinese medicine group compared with exercise group and western medicine group compared with exercise group had statistical significance (P<0.01).Traditional Chinese medicine group compared with western medicine group had no significance (P>0.05). (2) For the FS, traditional Chinese medicine combined exercise group increased most significantly and had statistical significance compared with traditional Chinese medicine group, western medicine group and exercise group (P<0.01). Traditional Chinese medicine group compared with western medicine group had no statistical significance (P>0.05). (3) For the SV, traditional Chinese medicine combined exercise group increase most significantly and had statistical significance compared with excercising training group. (4) For the LVESV, traditional Chinese medicine combined exercise group decreased most significantly and had statistical significance compared with the exercise group (P <0.01). (5) For the LVEDV, traditional Chinese medicine combined exercise group decreased most significantly and had statistical significance compared with Chinese medicine group, western medicine group and exercise training group (P <0.01).2. In model group, the MMP-1 mRNA expression increased significantly and had statistical significance compared with the sham operation group (P<0.01). After drugs treatment and (or) exercise training, the MMP-1 mRNA expression at the treatment group decrease significantly and had statistical significance compared with the model group (P<0.01). Chinese medicine combined exercise group decreased most significantly, followed by the Chinese medicine group, medicine group and exercise group. Traditional Chinese medicine combined exercise group had statistical significance compared with the Chinese medicine group, western medicine group and exercise group (P<0.01). Traditional Chinese medicine group and western medicine group had statistical significance compared with exercise group. Traditional Chinese medicine group had statistical significance compared with and western medicine group (P<0.05).3. In model group, the MMP-1 protein expression increased significantly and had statistical significance compared with the sham operation group (P<0.01). After drugs treatment and (or) exercise training, the MMP-1 mRNA expression at treatment group decreased significantly compared with model group (P<0.01). Chinese medicine combined exercise group decrease most significantly, followed by the western medicine group, traditional Chinese group and exercise training group. Traditional Chinese medicine combined exercise group had statistical significance compared with the Chinese medicine group, western medicine group and exercise group (P<0.01). Traditional Chinese medicine group and western medicine group had statistical significance compared with exercise group. Traditional Chinese medicine group had statistical significance compared with and western medicine group (P<0.05).4. In model group, the TIMP-1 mRNA expression decreased significantly and had statistical significance compared with sham control group (P <0.01); After drugs treatment and (or) exercise training, TIMP-1 mRNA expression at treatment group increased significantly compared with model group (P<0.01). Chinese medicine combined exercise group increased most significantly, followed by traditional Chinese group, western medicine group and exercise training group. Chinese medicine combined exercise group had statistical significance compared with western medicine group and exercise training group (P<0.01).Traditional Chinese medicine and western medicine group had statistical significance compared with exercise training group (P<0.01).Traditional Chinese medicine had statistical significance compared with western medicine (P < 0.05).5. In model group, the TIMP-1 protein expression decreased significantly and had statistical significance compared with sham control group (P <0.01); After drugs treatment and (or) exercise training, TIMP-1 protein expression at treatment group increased significantly compared with model group (P<0.01). Chinese medicine combined exercise group increased most significantly, followed by traditional Chinese group, western medicine group and exercise training group. Chinese medicine combined exercise group had statistical significance compared with westen medicine group and exercise training group (P<0.01). Traditional Chinese medicine group and western medicine group had statistical significance compared with exercise group (P<0.01). Traditional Chinese medicine had statistical significance compared with western medicine(P<0.01).6. In model group, the collagenⅠmRNA expression decreased significantly and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training,Ⅰcollagen mRNA expression slightly decreased in western group and exercise training group and had no statistical significance compared with model group(P>0.05). TheⅠcollagen mRNA expression increased significantly in Chinese medicine combined exercise group and traditional Chinese medicine group. Chinese medicine combined exercise group had statistical significance compared with exercise training and western medicine group(P<0.01). Taditional Chinese medicine had statistical significance compared with westen medicine group and exercise training group (P<0.01).7. In model group, the collagenⅠprotein expression decreased significantly and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training,Ⅰcollagen protein expression increased significantly at every treated group. Chinese medicine combined exercise group increased most significantly, followed by traditional Chinese group, western medicine group and exercise training group. Chinese medicine combined exercise group had statistical significance compared with traditional Chinese mecicine group ,western medicine group and exercise training group (P<0.01). Traditional Chinese medicine and western medicine group had statistical significance compared with exercise training group (P<0.01).8. In model group, the collagenⅢmRNA expression decreased significantly and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training,Ⅲcollagen mRNA expression increased significantly at every treated group. Westen medicine group increased most significantly, followed by Chinese medicine combined exercise group,traditional Chinese group and exercise training group. Westen medicine group had statistical significance compared with traditional Chinese mecicine group and exercise training group (P<0.01).Chinese medicine combined exercise group had statistical significance compared with Chinese mecicine group and exercise training group (P<0.01).9. In model group, the collagenⅢprotein expression decreased significantly and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training,Ⅲcollagen protein expression increased significantly at every treated group. Chinese medicine combined exercise group increased most significantly, followed by westen medicine group,traditional Chinese medicine group and exercise training group.combined exercise group,traditional Chinese group and exercise training group. Chinese medicine combined exercise group had statistical significance compared with traditional Chinese mecicine group,Westen medicine group and exercise training group (P<0.01). Traditional Chinese medicine and western medicine group had statistical significance compared with exercise training group (P<0.01).Western medicine group had no statistical significance compared with Chinese mecicine group(P>0.05).10.HE Histological observation showed that in the model group the myocardial cells were intertwined and had no stripes and out of shape. The myocardial nuclear were pyknosis.There were some white space and the heterochromatin were lumpish. Adjacent cells intercalated disk space widened and the tight junctions connecting capsules disappeared.The myofibril were deranged, broken and lossed. Morphology of myocardial cells in all treatment groups compared with the model group were improved, traditional Chinese medicine combined exercise group improved most significantly.11. Using immunohistochemical, in model group the MMP-1 average gray value was 550.14±132.83 and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training, the MMP-1 average gray value decreased significantly at every treated group.Chinese medicine combined exercise group decreased most significantly, followed by western medicine group, traditional Chinese group and exercise training group. Traditional Chinese medicine combined exercise training group ,traditional Chinese group and western medicine group had statistical significance compared with exercise training group (P<0.01).12. Using immunohistochemical, in model group the TIMP-1 average gray value was 115.83±43.25 and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training, the TIMP-1 average gray value increased significantly at every treated group.Westen group increased most significantly, followed by traditional Chinese medicine group, Chinese medicine combined exercise group and exercise training group. Traditional Chinese medicine combined exercise training group ,traditional Chinese group and western medicine group had statistical significance compared with exercise training group (P<0.01).13. Using immunohistochemical, in model group the the collagenⅠaverage gray value was 45.34±8.10 and had statistical significance compared with sham control group (P<0.01); After drugs treatment and (or) exercise training, the the collagenⅠaverage gray value increased significantly at every treated group. Chinese medicine combined exercise group increased most significantly, followed by traditional Chinese medicine group, western medicine group and exercise training group.Traditional Chinese medicine combined exercise group had statistic significance compared with western medicine group,traditional chinesee medicine group and exercise training group(P<0.01). Traditional Chinese medicine group and western medicine group had statistic significance compared with exercise training group(P<0.01).14. Using immunohistochemical, in model group the the collagenⅢaverage gray value was 61.46±7.11 and had statistical significance compared with sham control group (P <0.01); After drugs treatment and (or) exercise training, the the collagenⅢaverage gray value increased significantly at every treated group. Chinese medicine combined exercise group increased most significantly, followed by western medicine group, traditional Chinese medicine group and exercise training group. Chinese medicine combined exercise group had statistic significance compared with traditional western medicine group, traditional Chinesee medicine group and exercise training group(P<0.01). Traditional Chinese medicine group and western medicine group had statistic significance compared with exercise training group(P<0.01). Conclusion:1. By echocardiography,the behavior change in rats and myocardial tissue morphology observations,it proved that ligated coronary artery with exhaustive style swimming and hunger mehtods can successfully establish an animal model of chronic heart failure.2. Benifiting qi and actvating bloos compound recipe allying exercise training can increase the level of EF, SV, FS and decrease the level of LVEDV, LVESV in CHF rats,which will protect the heart function and inhibit the heart chamber to expand.3. Left ventricular remodeling in CHF rats is related to the MMP-1 protein and gene expression increase, TIMP-1 protein and gene expression decrease, MMP-1 / TIMP-1 ratio increase,ⅠandⅢcollagen decrease in cardiac tissue.4. Benifiting qi and actvating bloos compound recipe allying exercise training can regulate the collagen content and reduce the Left ventricular remodeling by decreasing the MMP-1 protein and gene expression, increasing the TIMP-1 protein and gene expression and decreasing the ratio of MMP-1/TIMP-1.5. Benifiting qi and actvating bloos compound recipe allying exercise training can protect heart function, reduce ventricular remodeling, which is better than traditional Chiese medicine group, exercise training group and western medicine group. It provides scientific basis and effective way for the rehabilitation of chronic heart failure. |
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