Study On The Mechanism Of Mechanism Of Moxibustion Combined With Benazepril On Endoplasmic Reticulum Stress-mediated Apoptosis In CHF Rats

Objective:To observe the effects of moxibustion combined with benazepril on endoplasmic reticulum stress-mediated apoptosis in rats with chronic heart failure(CHF),and explore the potential mechanism of moxibustion combined with Benazepril in the prevention and therapy of CHF,in order to offer more theoretical basis for clinical treatment of CHF.Method:A total of 60 SPF male SD rats were stochastically divided into blank series(n=10)and modeling series(n=50)based on random number table method after one week of adapted feeding.Rats in the blank series were not intervened,and the rats in the model series were treated with doxorubicin(DOX)by intraperitoneal injection to replicate the CHF model.DOX powder was diluted with 0.9% sodium chloride solution(1mg/ml),and the rats were injected intraperitoneally at a dose of 2.5mg/kg,and the cumulative injection dose was 15 mg/kg once a week for 6 consecutive weeks.After the modeling,the rats were fed for 3 consecutive weeks,and the model was identified by the general behavioral signs of the rats and echocardiography.The rats that did not die after modeling were stochastically divided into model series,moxibustion series,drug series(benazepril)and moxa series(moxibustion + benazepril),with 10 rats in each series.The rats in blank series and model series were anchored on the platform for 20 min during the intervention period.Rats in moxibustion series were shaved by shaving machine at acupoints and tied to the rat table."Fei Shu" and "Xinshu" were taken from both sides of the rats,and moxa sticks were placed 1-2cm above the two acupoints for gentle moxibustion for 20 min.The rats in the drug series were given benazepril hydrochloride suspension(0.86mg/kg).Rats in Moxa series were first given Benazepril hydrochloride suspension(0.86mg/kg),then moxibustion at "Feishu" and "Xinshu" points,moxibustion method was the same as that in moxibustion series.Except blank series and model series,other Seriess were treated once a day for 3consecutive weeks.After the intervention,the general behavioral signs of rats in each series were observed,and echocardiography was performed to detect ejection fraction(EF)and left ventricular diameter shortening rate(FS).Hematoxylin-eosin(HE)staining was used to observe the pathological changes of myocardial tissue.Serum B type brain natriuretic peptide(BNP)and angiotensin Ⅱ(AngⅡ)were measured by enzyme-linked immunosorbent assay(ELISA).The measured levels of CHOPmRNA and ATF4 mRNA in myocardium were detected by RT-qPCR.The expression levels of GRP78 and Caspase12 in myocardial tissue were detected by Western blot.Results:Compared with the blank Series,the rats in the modeling Series showed yellowing hair color,decreased diet,lethargy,weight loss,decreased activity,decreased urine volume,accompanied by asthma and fatigue,and both EF and FS values decreased in the experimental series(P<0.01).Significantly lower EF and FS for the model series compared to the blank series(P<0.01),serum BNP and AngⅡ contents were increased(P<0.01),myocardial cells were disordered,cell swelling,vacuole,inflammatory infiltration and other changes appeared in different degrees,and the nuclei of the cells were also significantly reduced.The expression levels of CHOPmRNA,ATF4 mRNA,GRP78 and Caspase12 increased significantly(P<0.01).Compared with model Series,EF and FS in treatment Seriess were increased(P<0.01),the contents of BNP and AngⅡin serum were decreased(P<0.01),the degree of pathological injury was reduced,the cell swelling and vacuol-like change were reduced,and the nucleus was increased.The expression levels of CHOPmRNA,ATF4 mRNA,GRP78 and Caspase12 were decreased(P<0.01).Compared with moxibustion Series,EF and FS in moxibustion Series and drug Series were further decreased(P<0.01,P<0.05),and the contents of BNP,AngⅡ,CHOPmRNA,ATF4 mRNA,GRP78 and Caspase12 were increased(P<0.01).Conclusion:1.Moxibustion can improve the general behavioral signs and cardiac function of CHF rats,reduce the levels of BNP and AngⅡ,reduce myocardial injury in rats,and delay the onset of CHF.2.Moxibustion,medication and moxibustion combined with medication can improve the cardiac function of CHF rats,reduce the levels of BNP and AngⅡ,and reduce myocardial injury in rats,and have good curative effects on CHF.The efficacy of moxibustion combined with medication in the treatment of CHF is better than that of single moxibustion or single medication.3.Moxibustion of bilateral "Feishu" and "Xinshu" points may improve CHF by inhibiting endoplasmic reticulum stress-mediated apoptosis-related ATF4 mRNA and CHOPmRNA relative expression and reducing the expression levels of related proteins GRP78 and Caspase12.