| Sig-1R acts as an important chaperone protein of the endoplasmic reticulum,and regulates Ca2+ transport in the mitochondria of the endoplasmic reticulum through interaction with IP3R,protects mitochondrial structure,promotes ATP production,increases SERCA2a activity,and improves Ca2+ cycling and energy metabolism disorders.The role of cardiomyocyte protection is the key to determining cell survival.It plays an important role in inhibiting compensatory hypertrophy of cardiac cells,reversing ventricular remodeling,and alleviating heart function.Based on the understanding of heart failure in Chinese medicine,the key to the pathogenesis of heart failure and blood stasis is the effect of Yiqi Huoxue Prescription(YQHXP)in the treatment of heart failure.Taking Sig-1R as an entry point to study the molecular mechanism of YQHXP in the treatment of cardiovascular diseases will open up new ideas for the treatment of heart failure at the subcellular level.Objective:From the perspective of subcellular organelles and their interrelatedness,we observed the effect of YQHXP,which improves myocardial energy metabolism,on the expression of Sig-1R and related molecules SERCA2a and IP3R,and the mitochondrial morphology,function,and energy metabolism of myocardial tissue.To further explore the mechanism of YQHXP in the treatment of heart failure by Sig-IR regulating endoplasmic reticulum/mitochondrial Ca2+ transport,improving energy metabolism and protecting cardiomyocytes from the perspective of subcellular organelles and their interactions.The theory of pathogenesis of blood stasis provides scientific and objective experimental evidence for the clinical application of prevention and treatment of heart failure.Methods:The model of heart failure after acute myocardial infarction was prepared by ligation of the left anterior descending coronary artery with SD rats,randomly divided into model group,low-dose YQHXP(YQHXP-L)group,high-dose YQHXP(YQHXP-H)group and fluvoxamine group.After 8 weeks of continuous administration of the drug,to assess heart function by rat cardiac ultrasound and body mass ratio measurements.HE staining and electron microscopy were used to observe cardiomyocytes and mitochondrial histomorphology.The expression of Sig-1R,SERCA2a,IP3R mRNA and protein were detected the myocardial infarction area by real-time fluorescence quantitative PCR,Western blot and ATP fluorescence intensity method was used to detect the change of ATP content in myocardial tissue of rats in each group.Results:1.After 8 weeks,echocardiography was used to examine the structure of the heart and its changes in systolic and diastolic function.The results showed that the rat ventricular structure was remodeled after 8 weeks.Compared with the sham group,left ventricular end-diastolic diameter,left ventricular end-systolic diameter,left ventricular end-diastolic volume,and left ventricular end-systolic volume were significantly increased(P<0.01).Left ventricular posterior wall contraction thickness and left ventricular posterior diastolic thickness were significantly reduced(P<0.01),the difference was statistically significant;and each drug group rats on the above indicators of myocardial remodeling Compared with the model group,there was a slight decrease(P<0.01).2.The results of echocardiographic functional indicators in each group showed:Compared with the sham group,the left ventricular ejection fraction and left ventricular short axis shortening rate of the model group rats were significantly reduced(P<0.01),and each drug group can significantly improve the heart EF%,FS%(P<0.01).3.The heart quality index of rats in the model group was significantly increased compared with the sham group(P<0.01).Compared with the model group,the heart quality index of the YQHXP-L group,the YQHXP-H group and the fluvoxamine group were significantly lower(P<0.01).4.HE staining of myocardial tissue in each group was performed under 20x magnification light microscope to observe the left ventricular infarction marginal zone.In the sham-operated group,the myocardial cells of the sham-operated rats were arranged neatly and orderly,the texture was clear,the cytoplasm was evenly colored.In the model group,the myocardial cells were arranged disorderly,the texture was disordered,the cytoplasm was unevenly stained,the nucleus was pyknosis,the myocardium fibers were broken,the structure was destroyed.The YQHXP-L group and YQHXP-H group were similar to the model group,but the lesion range was reduced.The pathological changes in the fluvoxamine group were similar to those in the model group.Disordered cell arrangement,uneven cytoplasm staining and nuclear pyknosis were also observed.5.Effect of YQHXP on Sig-1R,SERCA2a and IP3R mRNA expression in the border area of myocardial Infarction:Compared with the sham group,the expression of Sigma-1R mRNA in the myocardial tissue of the model group was decreased(P<0.05).The expression of Sigma-1R mRNA in the YQHXP-L,YQHXP-H group and fluvoxamine group was significantly higher than that in the model group(P<0.01);The expression of SERCA2a mRNA in the model group was lower than that in the sham group(P<0.05).Compared with the model group,the expression of SERCA2a mRNA was increased in the YQHXP-L group,there was no statistical difference(P>0.05);The expression of SERCA2a mRNA was significantly increased in the YQHXP-H group(P<0.01).The SERCA2a mRNA expression was also increased in the fluvoxamine group compared with the model group(P<0.05);The expression of IP3R mRNA in the model group was higher than that in the sham group(P<0.01).Compared with the model group,the expression of IP3R mRNA in the YQHXP-H group and YQHXP-L group was significantly lower(P<0.05).Compared with fluvoxamine group,the expression of IP3R mRNA was significantly decreased(P<0.01).6.Effect of YQHXP on expression of Sig-1R,SERCA2a and IP3R Proteins in the Border Area of myocardial infarction:Compared with sham operation,the expression of Sig-1R in myocardial tissue of the model group was significantly decreased,there was a statistical difference between the two(P<0.01);The Sig-1R levels in the YQHXP-L group,YQHXP-H group,and fluvoxamine group were higher than those in the model group,the difference was statistically significant(P<0.05).The expression of SERCA2a in the model group was lower than the sham group,at the ?=0.05 level.the difference was not statistically significant;SERCA2a is increased in YQHXP-L and fluvoxamine groups compared with the model group(P>0.05);Compared with the model group,the YQHXP-H group is reduced(P>0.05).The expression of IP3R in the model group was significantly higher than that in the sham group(P<0.05).Compared with the model group,the expression of IP3R was decreased in YQHXP-L group(P<0.05),and the expression of IP3R in the YQHXP-H group was significantly lower(P<0.0 1).the expression of IP3R in the fluvoxamine group was decreased,but there was no statistical difference(P>0.05).7.Effects of YQHXP on Myocardial Mitochondria Morphology after 8 Weeks of Operation:In the sham operation group,the myocardial fibers were arranged neatly and without fracture,and the stripes were clearly visible.The sarcomere is intact and the cytoplasm contains abundant mitochondria,which are round or oval.The mitochondria had normal morphology,no swelling,continuous and complete membrane,clear structure,orderly arrangement,and no vacuolization in mitochondria.Myocardium cells in the model group were disrupted,disorganized or even dissolved,with mitochondrial morphology varying in size,mitochondria swollen,destructive and unclear structure of iliac crest,increased interstinal matrix density,incomplete mitochondrial membrane,and partial vacuolization.;Compared with the model group,the YQHXP-L group,YQHXP-H group and the fluvoxamine group had a more orderly arrangement of myocardial fibers with less pathological changes,and the mitochondria were arranged in a regular,round,oval shape with clear and regular arrangement.8.Effect of YQHXP on ATP Content of Rat Myocardial Infarction Margin:The ATP concentration in the model group was lower than the sham group,with significant statistical difference(P<0.01);Compared with the model group,the ATP concentration in the YQHXP-L group and fluvoxamine group increased(P<0.05);The concentration of ATP in YQHXP-H group was significantly higher than that of the model group,and the difference was statistically significant(P<0.01).Conclusion:YQHXP-L and YQHXP-H can reduce inflammatory cell infiltration,improve cardiac morphological and functional indexes of rats,reverse ventricular remodeling and improve cardiac function in different degrees,and the effect is obvious.Its mechanism of action may be related to promoting the expression of Sig-1R,SERCA2a.At the same time,it inhibits the expression of IP3R.restoring mitochondrial morphological structure and function,and improving myocardial energy metabolism disorder.However,signal transduction between subcellular organelles is a complex network structure.Further research is needed on the more in-depth molecular mechanisms of Sig-1R in the protection of cardiomyocytes and the effects of YQHXP. |
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