| Objective:By using adriamycin intraperitoneal injection method to induce the DCM rat model. By series intraperitoneal injection to detecte the expression of phospholamban(PLN) and sarcoplasmic reticulum calcium ATP enzyme(SERCA2a) in an adriamycin-induced cardiomyopathy rat model,and determine the expression of DCM myocardial PLN and SERCA2a after using astragalus and carvedilol to improve the cardiac function, and seek the relationship between the PLN, SERCA2a and left ventricular ejection fraction(LVEF).Method:82 Wistar rats were randomly assigned to normal group(n=10) and DCM model group(n=72). Injected adriamycin 2mg/kg/times to the abaoinal cavity in the model group, and the same quantity with saline to normal control group. Both groups were injected 1 week and 8 weeks continuously. After successful modeling, LVEF of 5 rats were measured. Divide the 40 DCM model rats randomly into DCM group(n=14), asragus group(n=13) and carvedilol group(n=13), meanwhile, observe the 5 rats in the normal group. A drug interference to the four groups as following methods:fill the stomaches with 2ml saline in both DCM group and the normal control group; fill the stomaches with 2ml saline and astragalus grain of 1 g/kg/d(Astragaloside 0.6mg/kg/d) in the astragalus group; fillthe the stomaches with 2ml saline and carvedilol of 10mg/kg/d in the carvedilol group. Four groups were 1 times/day for 4 weeks. At 12 weeks, 5 rats in each group were taken and detected LVEF and then killed, leaving the left ventricle of rats of the 8th week and 12th week for PLN, SERCA2a RT-PCR and Western-Blot.Results:1. The model of DCM rats were successfully established by intraperitoneal injection adriamycin eight weeks weekly.2. The expression of PLN in DCM rats:At 8 weeks and 12 weeks, compared with normal rats respectively, the expression of PLN at mRNA and protein levels were significantly higher(.P<0.05). Compared with 8 weeks and 12 weeks of DCM rats, the expression of PLN at mRNA and protein levels were no significant changed (P>0.05).3. The relationship of PLN and SERCA2a in DCM rats:Compared with the same age in normal rats, the expression of SERCA2a mRNA and protein levels in 8 and 12 weeks DCM rats were significantly lower(P<0.05). The scatter were drawn and the linear correlation analysis were given through SERCA2a mRNA and protein levels in 12 weeks rats, shown a negative correlation, r values are-0.99,-0.89, P<0.05.4. The relationship between PLN and LVEF in DCM rats:Compared with 8 and 12 weeks DCM rats, the LVEF of 12 weeks was significantly decreased, P<0.05. The scatter were drawn and the linear correlation analysis were given through PLN mRNA and protein levels respectively in 12 weeks rats with LVEF, shown a negative correlation, r values are-0.965,-0.925, P＜0.05.5. The relationship between SERCA2a and LVEF in DCM rats:The scatter were drawn and the linear correlation analysis were given through SERCA2a mRNA and protein levels respectively in 12 weeks rats with LVEF, shown a positive correlation, r value respectively is 0.93,0.996, P<0.05.6. Compared with 8 weekend DCM group, LVEF of astragalus group and carvedilol group were significantly increased. Compared with 8 and 12 weekend DCM group respectively, the expression of PLN at mRNA and protein levels of astragalus group and carvedilol group were significantly decreased, and SERCA2a were increased(P<0.05).Conclusion:1. The expression of PLN in DCM group was increased, but over time,there were no significant changes; at the same time, the expression of SERCA2a was reducted and LVEF was declined.2. The expression of myocardial PLN in DCM group was negatively correlated with LVEF and SERCA2a, and SERCA2a was positively related with LVEF.3. After LVEF of DCM rats was improved by astragalus and carvedilol,the expression of PLN was significantly decreased, and the expression of SERCA2a was increased.4. The mechanism of PLAN affect LVEF was primarily associated with decreased expression of PLN and enhanced SERCA2a. |
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