Efficacy And Mechanism Of Yiqiwenyang Formula In The Prevention And Treatment Of Chemotherapy-induced Cardiotoxicity

Although immune,targeted and other tumor treatment methods have made breakthroughs,chemotherapy is still one of the cornerstones of anti-cancer treatment in medical oncology.How to reduce the side effects of chemotherapy is still the current issues.Chemotherapy drugs can not only kill tumor cells,but also damage normal cells and tissues,resulting in toxic side effects such as cardiotoxicity,gastrointestinal reactions,bone marrow suppression,neurotoxicity and so on.Among them,the harm of heart disease related to chemotherapy drugs has become increasingly prominent,and it has become the leading cause of non-cancer-related deaths in cancer patients.Doxorubicin(Dox)is one of the representative anthracycline drugs with the highest incidence and severity of cardiotoxicity.The current research mechanisms mainly focus on oxidative stress injury,autophagy,and apoptosis mechanisms.In terms of treatment,only dexrazoxane is proved by FDA to prevent cardiotoxicity.Therefore,it is still urgent to explore drugs that can protect the heart without affecting the antitumor efficacy.Traditional Chinese medicine(TCM)is effective in reducing cardiotoxicity of chemotherapy,which can reduce the occurrence of cardiotoxicity,delay the progression of cardiotoxicity and improve the quality of patients’ life.When summarized the experience of Professor Li Peiwen,a national famous Chinese medicine professor,we found that Yiqiwenyang Formula could improve the patient’s tolerance to chemotherapy by increasing heart rate variability(HRV)and inhibiting the expression of inflammatory factor.The purpose of this study is to explore the efficacy and mechanism of Yiqiwenyang Formula in the prevention and treatment of chemotherapy cardiotoxicity,the value of HRV in the diagnosis of chemotherapy cardiotoxicity.Objective:1.To explore the curative effect and mechanism of Yiqiwenyang Formula through the SD rats’ model of Dox cardiotoxicity.2.To explore the diagnostic significance of HRV for chemotherapy cardiotoxicity through the analysis of heart rate variability.3.To analyze the academic point of view of "Treatment of adverse reactions of chemotherapy from the heart" by summarizing relevant literature on adverse reactions of chemotherapy.Methods:1.Using the method of network pharmacology to preliminarily analyze the mechanism of Yiqiwenyang Formula in preventing and treating cardiotoxicity of chemotherapy,and provide directions for the development of basic experiments.2.To construct the SD rats’ models to evaluate the effects of Yiqiwenyang Formula in preventing and treating cardiotoxicity through the changes in rat body weight,myocardial enzymes,cardiac function,and apoptosis of myocardial cells.4.ELISA,Western blotting and other techniques were used to clarify the mechanism of Yiqiwenyang Formula to reduce the cardiotoxicity of Dox in terms of oxidative stress and mitochondria-related endogenous apoptosis pathway.5.Through the analysis of HRV in SD rats of Labchart software,the correlation analysis was carried out with cardiac function indexes to evaluate whether it could be used as the diagnostic index of chemotherapy cardiotoxicity.6.Establish mouse-derived 4T1 and human-derived MDA-MB-231 breast cancer cell models to explore the effect of Yiqiwenyang Formula on the antitumor activity of Dox.7.Consult the basic theory of TCM and clinical experience to analyze Professor Zhu Shijie’s academic thoughts on "Treatment of side effects of chemotherapy from the heart".Results:1.Network pharmacology analysis:Yiqiwenyang Formula can protect the heart by interfering apoptosis pathway with an effect on Caspase3,BCL-2 protein through analysis of core target genes and KEGG.2.Yiqiwenyang Formula can reduce the cardiotoxicity of Dox:The acute myocardial injury model was established by intraperitoneal injection of doxorubicin 15 mg/kg in SD rats.The results showed that Dox could induce cardiac injury in SD rats,resulting in the increase of serum cTnI and BNP(P<0.05).Dox can also decrease the indexes of cardiac ejection function LVEF and FS(P<0.05).Myocardial cells showed pathological changes such as coagulation necrosis and cytoplasmic lysis,and the proportion of myocardial cell apoptosis increased(P<0.05).Both the high-dose and middle-dose groups of Yiqiwenyang Formula can significantly improve the weight loss of rats caused by Dox,reduce the contents of cTnI and BNP in serum(P<0.05),improve cardiac function,reduce cardiac pathological damage and myocardial injury.The proportion of apoptotic cells(P<0.05)showed no significant difference between the two groups(P>0.05).However,in the experimental process,we found that the appetite of the rats in the high-dose Yiqiwenyang Formula group was significantly reduced,and the body weight showed a downward trend compared with the middle-dose group.Therefore,in the following research,the middle-dose group of Yiqiwenyang Formula can be used to explore the mechanism.3.The machinimas of Yiqiwenyang Formula in reducing cardiotoxicity induced by Dox:Combined with the previous network pharmacology analysis,the protective mechanism of Yiqiwenyang Formula on the heart was explored from the direction of oxidative stress injury and cardiomyocyte apoptosis pathway.Studies have shown that,compared with the normal group,the production of ROS and MDA was significantly increased(P<0.05),the activities of SOD and GSH-Px were significantly reduced(P<0.05).The levels of pro-apoptotic factor Cyt-C,Caspase3 and Caspase9 were upregulated(P<0.05),and the ratio of Bax/Bcl-2 was increased(P<0.05).However,Yiqiwenyang Formula can reduce the contents of oxidative stress products ROS and MDA(P<0.05),increase the activities of antioxidant enzymes GSH-Px and SOD(P<0.05),and inhibit the apoptosis factor Cyt-C and apoptosis.The protein levels of apoptosis proteins Caspase3 and Caspase9(P<0.05),and down-regulation of Bax/Bcl2(P<0.05)protected the heart.4.HRV can be used to diagnose cardiotoxicity:HRV index can reflect the overall state of cardiac autonomic nerve function,and can be divided into time domain index and frequency domain index according to different algorithms.The time domain index SDNN in Dox group was significantly lower than the normal group(P<0.05).The correlation analysis between SDNN and the indicators of cardiac function showed that SDNN was positively correlated with LVEF(r=0.8783,P<0.001)and FS(r=0.8352,P<0.001),negatively correlated with serum BNP level(r=-0.9365,P<0.05).SDNN decreases with decreased cardiac function in a cardiotoxic state.The frequency domain index showed that HF decreased,LF increased,and LF/HF increased in the Dox group(P<0.05).It shows that the sympathetic nerve is activated and parasympathetic nerve is reduced,the balance of them is disturbed.Compared with the Dox group,Yiqiwenyang Formula can increase SDNN and HF,decrease LF and HF/LF ratio(P<0.05),which means that Yiqiwenyang Formula increase HRV and regulate cardiac autonomic nerve function.5.Yiqiwenyang Formula has no effect on the anti-tumor effect of Dox:The key to the prevention and treatment of chemotherapeutic cardiotoxic drugs is to find drugs that protect the heart without affecting the anti-tumor efficacy of doxorubicin.Vitro studies showed that when combine Yiqiwenyang Formula and doxorubicin,compared with the doxorubicin single application group,there was no significant difference in the activity of 4T1 cells and MDA-MB-231 cells(P>0.05).Yiqiwenyang Formula has no effect on the anti-tumor effect of Dox.6.Preliminary formation of the theory of treating side effects of chemotherapy from the Heart:The symptoms such as fatigue,shortness of breath,palpitation,insomnia are very common with chemotherapy patients in the early stage,which are performance of heart qi deficiency and heart spirit disturbation.It can lead to a series of diseases such as damage to heart yang and blood stasis,affecting the physiological functions of the heart storing spirit and controlling blood vessels.Therefore,the overall function of the "heart" should be adjusted to receive better curative effect.Conclusions:1.Yiqiwenyang Formula has a certain curative effect on Dox induced cardiotoxicity.The mechanism is to decrease oxidative products,increase the activity of antioxidant enzymes,and protect the myocardium by regulating the mitochondriamediated endogenous apoptosis pathway.3.Chemotherapy heart injury can lead to decrease in HRV.Yiqiwenyang Formula can increase HRV.HRV can be used to diagnose cardiotoxicity.4.The adverse reactions of chemotherapy are closely related to TCM theory "the heart stores the spirit and controls the blood vessels".The academic point of view of treating the adverse reactions of chemotherapy from the heart has initially formed,which has guiding value for clinical application.