Mechanism of action of Xuefu Zhuyu Decoction on model rats of coronary heart disease with blood stasis pattern based on mitochondrial calcium homeostasis
Objective To explore the mechanism of action of Xuefu Zhuyu Decoction(XFZYD)in treating coronary heart disease(CHD)with blood stasis pattern based on mitochondrial calcium homeostasis.Methods A total of 150 SD rats were randomized into normal group,sham-operated group(only threading without ligation),model group,atorvastatin calcium group(0.90 mg·kg-1·d-1,hereinafter referred to as statin group),and low-,medium-,and high-dose XFZYD groups(3.51,7.02,14.04 g·kg-1·d-1,hereinafter referred to as low-,medium-,and high-dose groups),with six rats in each group.Except for the normal and sham-operated groups,the other groups were prepared for CHD with blood stasis pattern models by ligating the left anterior descending coronary artery of the rats.After 24 hours of successful modeling,the rats were given continuous intragastric administration for 7 d.The myocardial tissue of left ventricular ischemic area in rats of each group was taken.HE staining was used to observe the necrosis of the samples.The fluorescence intensity of calcium ions in mitochondria was determined by mitochondrial calcium ion probe.The Ca2+-Mg2+-ATPase activity of myocardial cells was examined by phosphorus determination method.Immunohistochemistry was used to check the protein expressions of mitochondrial calcium uniporter(MCU),sarco/endoplasmic reticulum Ca2+-ATPase 2a(SERCA2a),transient receptor potential canonical 3(TRPC3),calcium release-activated calcium channel protein 1(ORAI1),cysteinyl aspartate specific proteinase-3(Caspase-3),and cysteinyl aspartate specific proteinase-9(Caspase-9).Results Compared with the normal group,the model group showed disordered cell arrangement,blurred transverse stripes,necrosis of a large number of myocardial cells,a significant increase in mitochondrial Ca2+average fluorescence intensity and AOD values of MCU,TRPC3,ORAI1,Caspase-3,and Caspase-9(P＜0.01),and an obvious decrease in Ca2+-Mg2+-ATPase activity and AOD value of SERCA2a in the myocardial tissue of rats(P＜0.01).Compared with the model group,the statin group and low-,medium-,and high-dose groups showed reduced myocardial cell edema,inflammatory cell infiltration,and cell pathological damage as well as relatively tight cell arrangement,meanwhile,the AOD value of SERCA2a in those groups significantly increased(P＜0.01),and the average fluorescence intensity of mitochondrial Ca2+,AOD values of MCU,TRPC3,ORAI1,and Caspase-9 decreased significantly(P＜0.05,P＜0.01).In addition,the Ca2*-Mg2+-ATPase activity in low-,medium-,and high-dose groups increased significantly(P＜0.05,P＜0.01),while the AOD value of Caspase-3 decreased significantly(P＜0.05,P＜0.01).Conclusion XFZYD can regulate myocardial mitochondrial calcium ion transport and mitochondrial calcium homeostasis in rats of CHD with blood stasis pattern,which may be related to up-regulating the protein expression of SERCA2a and down-regulating the protein expressions of MCU,Caspase-3,and Caspase-9.
万方数据
维普
