| Objective: To investigate the effects of Water-soluble propolis( WSP) on myocardial cells apoptosis in rats with chronic heart failure and its possible mechanism.Methods: Randomly selected 6 male SD rats as normal control group( NC group), male SD rats modeling by intraperitoneal injection of adriamycin were randomly divided into five groups( n=6) : Chronic heart failure group( CHF group), WSP low dose group( LD group), WSP middle dose group( MD group), WSP high dose group( HD group), digoxin group( DIG group). Rats in those groups were injected with adriamycin( 2mg.kg-1.w-1) for 6 weeks. After the third injection adriamycin,rats in LD, MD, HD and DIG groups were given total flavonoids of propolis in WSP 50mg/kg, 100mg/kg, 200mg/kg and digoxin 0.02mg/kg intragastrically, once a day for 4 weeks; at the same time, the rats in CHF and NC groups were given equal volume of saline gavage, the rats in NC group were given equal volume of saline by intraperitoneal injection for 6 weeks. After six weeks recorded the ECG and cardiac function index of rats by signal acquisition system; While calculated value of rat heart weight/body weight( HW/BW). Plasma cardiac troponin I( c Tn I), interleukin-6( IL-6), tumor necrosis factor-α( TNF-α), brain natriuretic peptide( BNP)content of rats detected by ELISA. Observing the rats in each group myocardial morphological changes and collagen fiber hyperplasia by HE and Masson staining, calculation of collagen volume fraction( CVF%); myocardial apoptosis was detected with Tunel method, and calculation of myocardial apoptosis index. P-Cx43 expression was detected by western blotting method. The relationship between P-Cx43 expression level and apoptotic index was analyzed by Pearson’s correlation.Results: During the experiment, we observed the general state of rats found that rats in NC group were no deaths, mental health was fine, eating and drinking were normal; Rats in CHF group showed weight loss, listlessness, eating less and activity reduced; The situation of WSP groups rats improved compared with CHF group. Compared with NC group, QRS interval of CHF was significantly widened(P<0.01), ST–T segment elevation obviously( P<0.01); LVSP, +dp/dtmax,-dp/dtmax in CHF group significantly lower( P<0.01), LVEDP increased significantly( P<0.01); Compared with CHF group, QRS interva were significantly smaller and ST–T segment decreased in WSP groups( P<0.01); LVSP, +dp/dtmax,-dp/dtmax in MD and HD groups increased( P<0.05), LVEDP decreased significantly in MD and HD groups( P<0.01), LVEDP in LD group obviously decreased( P<0.01), and LVSP, +dp/dtmax,-dp/dtmax in LD group change was not obvious( P>0.05). Compared with NC group, HW/BW of CHF group was hihger( P<0.05); plasma BNP, c Tn I, TNF-α and IL-6 content in the CHF group were significantly higher( P<0.01); Compared with CHF group, HW/BW of HD group decreased( P<0.05), HW/BW of LD and MD groups change was not obvious( P>0.05); BNP, c Tn I, TNF-α and IL-6 content in MD and HD groups were significantly reduced( P<0.01), those plasma indicators in LD group changes were not obvious( P>0.05). HE staining showed that myocardial fibers of rats in the NC group arranged in neat rows,myocardial cell size, normal morphology; Myocardial cells of rats in CHF group were arranged in disorder, with cytoplasmic dissolved state, hyperchromatic nuclei, myocyte gap visible myocardial fiber fracture, dissolution widened obviously; The situation of rats in LD, MD, HD groups improved compared with CHF group. Masson staining results showed there were more blue myocardial collagen deposition in the surrounding cardiomyocytes in the CHF group, CVF% of CHF group( 17.18+4.49) was higher than that in NC group( 5.57+3.57)(P<0.01); MD and HD groups CVF% respectively( 11.19+2.60),( 7.10+1.83) were reduced obviously compared with CHF group( P<0.01), LD group CVF%( 15.56+4.40) compared with the CHF group had no significant difference( P>0.05). The result of Tunel staining showed:In CHF group, myocardial apoptosis indec( %)( 22.62+3.39) than that in NC group( 1.12+0.24) increased significantly( P<0.01); Compared with CHF group, the apoptosis of myocardial cells index( %) of LD, MD and HD groups respectively( 15.79+2.8),( 9.28+2.1) and( 4.73+1.14) were significantly lower than those in CHF group( P<0.01). Western blotting showed that P-Cx43 expression in NC group was significantly lower than that in CHF group( P<0.01); And that in all WSP treatment groups were decreased compared with CHF group( P<0.05 or P<0.01), among which pairwise comparisons also showed differences( P<0.05). The expression level of P-Cx43 was positively correlated with the apoptotic index(r=0.861,P<0.01).Conclusion: Water-soluble propolis dose-dependent improvement in myocardial tissue morphology, inhibition of ventricular remodeling, improve cardiac function, delay myocardial cells apoptosis in rats of CHF, the possible intrinsic mechanism is closely related to the regulation of Cx43 expression. |
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