Effects Of Shen Qi Yangxin Decoction On High Mobility Group Box 1 Inflammatory Signal Pathway In Rats With Dilated Cardiomyopathy

Objective:To observe the effects of Shen Qi Yangxin Decoction(SQYXD)on survival rate、heart weight index、heart function and morphology of myocardial injury;investigate the effect of SQYXD on High Mobility Group Box 1 inflammatory signal pathways in rats with Adriamycin-inducing dilated cardiomyopathy(DCM),to explore its possible mechanism.Methods:DCM Models were repeated by Adriamycin(1.0mg/Kg,BIW,via intraperitonea)for 6 weeks(n=150),another rats served as normal group(n=10).By the end of 8 week 10 rats were randomly selected to see whether the model was successful.At the sametime,16 rats of were randomly selected as the baseline group,the echocardiography and 18F-FDG Micro-PET myocardial metabolism imaging were measured on them,and the serum and left ventricular myocardial tissue were collected from the rats.Then the remaining DCM model rats were randomly divided into 4 groups:Perindopril group(n=20),Metoprolol group(n=20),SQYXD group(n=20),and model group(n=23).Perindopril was administered by gavage with a dose of 3 mg/kg/d in Perindopril group for 8 weeks,metoprolol was administered by gavage with a dose of 50 mg/kg,bid,in Perindopril group for 8 weeks,SQYXD solution was administered by gavage with a dose of 1.87 g/kg/d in SQYXD group for 8 weeks,distilled water was administered in model group,normal for 8 weeks.The left ventricular dimension at end-diastole(LEVDD)and at end-systole(LVESD),left ventricle ejection fraction(LVEF)and fraction shortening(FS)were measured with echocardiography at the 9th week after gavaged,then rats serum levels of BNP、TNF-α、IL-1、IL-6、IL-10、CRP were measured by the method of enzyme-linked immunosorbent assay.Histopathological characteristics of left ventricular myocardial tissue were observed by HE and transmission electron microscopy.High Mobility Group Box 1(HMGB1),Receptor for advanced glycation end product(RAGE),Toll-like receptor-4(TLR-4),NF-κB mRNAs and Proteins in rats myocardial tissue were determined by the methods of RT-PCR and western blot.The 18F-FDG Micro-PET myocardial metabolism imaging was performed at the 9th week after gavaged in the group of SQYXD group,model group,and normal group.Results:(1)Treatment of DCM rats:1.The LEVDD and LVESD were significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=27.972 P<0.001;F=33.281 P<0.001).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.The LVEF and LVFS were significant higher in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=22.798 P<0.001;F=39.423 P<0.001).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.2.The serum levels of BNP was significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=24.840 P<0.001).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.3.Heart weight index(HWI):The HWI was significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=6.323 P<0.001).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.4.Haematoxylin and eosin staining:Myocardial samples from model groups showed extensive cardiomyocyt evacuolization,degeneration,and necrosis;interstitial edema;inflammatory cell infiltration;and replacement fibrosis.Myocardial samples collected from the Perindopril group,Metoprolol group and SQYXD group showed significantly improved in histopathological changes,as expected.But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.5.Microscopic observation:The fibril of cardiac muscle broke and mitochondria swelled;vacuolar degeneration and the inner membrane ridge disappearance in model group,myocardial samples collected from the SQYXD group showed significantly improved.But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.6.The myocardial glucose metabolism and myocardial 18F-FDG uptake was significantly reduced in the model group,The myocardial glucose metabolism and myocardial 18F-FDG uptake of the SQYXD group was significantly improved compared with model control group.(2)HMGB1 and its inflammatory signal pathways in DCM rats:The expression of HMGB1 mRNA、TLR-4mRNA、RAGE mRNA、NF-κB mRNA in myocardial tissue were significant higher in baseline group than in the normal group.The expression of HMGB1mRNA were positively correlated with TLR-4 mRNA,RAGE mRNA and NF-mRNA.(R=0.904,P<0.000;R=0.952,P<0.000;R=0.909,P=0.002).At the same time,the protein expression in myocardial tissue of HMGB 1、TLR-4、RAGE、NF-κB were also significantly increased in baseline group than in the normal group.The serum levels of TNF-α、IL-1.IL-6、IL-10 and CRP were significantly higher in baseline group.The expression of HMGB 1 mRNA in myocardial tissue was positively correlated with TNF-α、IL-1、IL-6、IL-10 and CRP(R=0.958,P=0.000;R=0.933,P=0.002;R=0.984,P=0.001;R=0.946,P=0.000;R=0.950,P=0.000).The expression of HMGB 1 mRNA was positive correlated with BNP and LVEDD(R=0.916,P=0.001)and it was negative correlated with LVEF(R=-0.880,P=0.001).(3)Treatment of HMGB 1 and its inflammatory signal pathways in DCM rats:1.The expression of HMGB 1 mRNA、RAGE mRNA、TLR-4mRNA、NF-κB mRNA in myocardial tissue were significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=5.684 P<0.001;F=3.597 P=0.009;F=5.215 P=0.001;F=4.705 P=0.002).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.2.The protein expression of HMGB 1、RAGE、TLR-4、NF-κB in myocardial tissue were significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group;The protein expression of HMGB1、RAGE、TLR-4、NF-κB in myocardial tissue were significant decreased in SQYXD group than in the Perindopril group and Metoprolol group.3.The serum levels of TNF-α、IL-1、IL-6、IL-10 were significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=9.365 P<0.001;F=15.387 P<0.001;F=8.490 P<0.001;F=7.400 P<0.001).But there’s no significant difference between Perindopril group,Metoprolol group and SQYXD group.4.The serum levels of CRP were significant decreased in Perindopril group,Metoprolol group and SQYXD group than in the model group(F=14.524 P<0.001).But there’s no significan difference between Perindopril group,Metoprolol group and SQYXD group.Conclusion:(1)HMGB1 and its inflammatory signaling pathways(HMGB1-TLR4/RAGE-NF κB-cytokine)participates in the occurrence and development of dilated cardiomyopathy.(2)Shen Qi Yangxin Decoction can effectively reduce the dilated left ventricular diameter,and improve heart function of dilated cardiomyopathy.(3)The mechanisms of Shen Qi Yangxin Decoction on DCM maybe regulate the gene and protein expression of HMGB1 and its inflammatory signal pathways in rats with dilated cardiomyopathy.