ETHNOPHARMACOLOGICAL RELEVANCE:"Qi deficiency and blood stasis" syndrome is one of the most common syndromes treated with Traditional Chinese Medicine among ischemic heart disease (IHD) patients in clinic. As a Chinese herbal formula with the function of tonifying Qi and activating blood, Yiqihuoxue Decoction (YQHX) has been frequently proven to be effective in the clinical treatment of IHD.AIM OF THE STUDY:The cardioprotective mechanisms of YQHX in treating ischemic heart disease were investigated, with emphasis on the key targets and pathways.MATERIALS AND METHODS:In the present study, the potential targets of compounds identified in YQHX were predicted using PharmMapper, Symmap, and STITCH databases, and a "herb-compound-target" network was constructed using Cytoscape. Subsequently, the GO and KEGG functional enrichment analyses were analyzed using the DAVID database. Furthermore, a protein-protein interaction network was constructed using STRING to obtain the key target information. Besides, we used a myocardial ischemia rat model to investigate the cardioprotective effects of YQHX. Transmission electron microscopy and Western blotting were used to observe apoptotic bodies and confirm protein expressions of key candidate targets, respectively.RESULTS:Network pharmacology showed that a total of 141 potential targets were obtained from these databases. The functional analysis results revealed that the targets of YQHX were largely associated with apoptosis, and the PI3K-AKT and MAPK pathways might represent key functional pathways. The hub genes of network include ALB, TP53, AKT1, TNF, VEGFA, EGFR, MAPK1, CASP3, JUN, FN1, MMP9, and MAPK8. In vivo, YQHX significantly improved cardiac function and suppressed apoptosis in ischemic rat myocardium. Furthermore, YQHX could significantly upregulate Nrf2 and HO-1 expression, and inhibit JNK phosphorylation.CONCLUSIONS:Based on network pharmacology and experimental evidence, this study proves that the cardioprotective effects and mechanisms of YQHX depend on multi-component, multi-target, and multi-pathway. In particular, YQHX exerts anti-apoptotic effects potentially by regulating the Nrf2/HO-1 and JNK-MAPK pathways.

Objective: To investigate the cardioprotective effect of Yiqi Huoxue granule (YQHXG) in the regulation of autophagy in rats induced with myocardial infarction (MI). Methods: An acute MI animal model was established by ligation of the left anterior descending branch of the coronary artery in Sprague-Dawley rats. Besides, 20 rats received sham operation were classified into a control group. The remaining 59 rats were randomly divided into MI model group (n = 19), YQHXG group (n = 20), and perindopril group (n = 20). Relevant indicators on days 7 and 28 were observed in each group. Left ventricular function was determined by echocardiography. The structure and morphology of mitochondria, and the number of autophagic vesicles, were observed by transmission electron microscopy. The mRNA and protein expression levels of LC3, FUNDC1, Beclin-1, and BNIP3 were examined in the tissue of the MI marginal area. Results: Compared with the MI model group, YQHXG showed obvious improvements in cardiac functions. Observing the microscopic morphology of the heart tissue, myocardial tissue damage attenuated, autophagic signs of autophagosomes and autolysosomes reduced, vacuolization in mitochondria mitigated, and mitochondria arranged in order. YQHXG could reduce the degree of tissue lesion after MI and regulate the expression of autophagy-related molecules at different stages. On Day 7, YQHXG significantly downregulated the expression of Fundc1, Becn1, Bnip3 mRNA and reduced the levels of FUNDC1, Beclin-1, BNIP3, and LC3 B proteins expression (all P < .001). On Day 28, YQHXG could upregulate the expression of Becn1, Fundc1 and Bnip3 mRNA and increased the levels of the corresponding proteins expression (all P < .001). Besides, it also increased LC3 B protein expression level (P = .0344). Conclusion: YQHXG regulated the expression of mitochondrial autophagy-related factors in myocardial tissue and mitochondrial autophagic activity at different stages to protect the heart following MI.

目的 观察动脉粥样硬化兔模型内皮细胞标志物的变化及调脂通脉中药的干预作用.方法 采用高脂饲料饮食和球囊损伤颈动脉的方法复制动脉粥样硬化兔模型,随机分为正常组、模型组、假手术组、西药组(阿托伐他汀,1.1 mg/kg)和中药组(调脂通脉中药,7.3 g/kg)组,共5组,选择治疗后第6周、12周为观察点.在两个时间点,检测血脂水平;HE染色检测颈总动脉、肝脏形态学变化;ELISA检测内皮细胞标志物内皮素(ET-1)、一氧化氮合酶(eNOS)、前列环素2(PGI2)、血栓素A2(TXA2)的变化.结果 两个时间点,模型组ET-1、TXA2表达增加(P＜0.05)、eNOS、PGI2表达下降(P＜0.05).调脂通脉中药能够降低ET-1、TXA2水平(P＜0.01),且升高eNOS、PGI2表达(P＜0.05).结论 调脂通脉中药能够改善血管内皮细胞损伤,减轻动脉粥样硬化.

Objective To explore the protective effects of Yiqi Huoxue Fang(Qi-boosting Blood-activa-ting Formula,YQHXF)on rat myocardium with myocardial infarction and its possible mechanism by ob-serving the effects of YQHXF on necrosis and apoptosis of myocardial cell necrosis.Methods Male SD rats were randomly divided into the sham-surgery group,model group,YQHXF group,and perindopril group.Animal model of myocardial infarction was built by ligating coronary artery on the left anterior de-scending branch.Subsequently,YQHXF group received intragastric administration of Yiqi Huoxue Fang (compound medicine,21 g/kg)once per day,and perindopril group was given perindopril (0.4 mg/kg)once a day;Sham-surgery group and model group received sterile distilled water without drug inter-vention.The blood in abdominal aorta was drawn,and the tissue on the peripheral area of the myocardial infarction was taken at two fixed time points (on the 7 th and 28 th day)after administration.Ultrasonic ech-ocardiography was used to detect changes on cardiac morphology and function in rats:left ventricular e-jection fraction (LVEF),left ventricular short axis ratio (LVFS),left ventricular end systolic diameter (LVIDs),left ventricular end-diastolic diameter (LVIDd);serum superoxide dismutase (SOD),gluta-thione peroxidase (gsh-px),creatine kinase (CK),lactate dehydrogenase (LDH),aspertate amin-otransferase (AST),and troponin (cTnT)were measured with ELISA method.Protein imprinting meth-od (Western Blot)was used to measure cytochrome C (CytC)and protein expression of cysteine aspartic acid and protease-3 (caspase-3)in the border zone of infarcted area.Results On the 7 th day,com-pared with the model group,LVEF and LVFS in YQHXF group and perindopril group decreased without statistical significance(P >0.05).On the 28 th day,compared with the model group,LVEF and LVFS in YQHXF group and perindopril group increased significantly(P >0.05 ).LVIDs and LVIDd decreased markedly(P <0.05);LVIDd in perindopril group decreased sharply (P <0.05),and decrease presen-ted on LVIDs without statistical significace (P >0.05).On the 7 th and 28 th day,compared with the mod-el group,CK-MB,LDH,AST,cTnT in YQHXF group and perindopril group decreased significantly (P<0.05),and SOD,GSH-PX increased significantly.On the 7 th day,protein expression of cytC,caspase-3 decreased significantly in YQHXF group and perindopril group(P <0.05).Conclusion Administra-tion of YQHXF at early stage can effectively improve myocardial infarction on cardiac structure and func-tion in rats with myocardial infarction,improve oxidation resistance of myocardial cell,and inhibit myo-cardial apoptosis and necrosis.

OBJECTIVETo investigate the effect of Yiqihuoxue prescription (YQHX) from Traditional Chinese Medicine (TCM) on myocardial glucose and lipid metabolism after myocardial infarction via the cross talk between the liver kinase B1 (LKB1)-dependent Notch1 and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK). YQHX was prepared with substances with properties that benefit, to activate blood circulation based on the TCM theory.METHODSAnimal models of myocardial infarction were established by ligating Sprague Dawley rats' left anterior descending coronary arteries. The animals were randomly divided into a myocardial infarction (MI) group, a YQHX group, a perindopril group, a r-secretase inhibitor, Notch signal inhibitor (DAPT) group, a DAPT+YQHX group and a sham group. The related drugs were administered on the second day after operation, and changes in the relevant indexes were examined on weeks 1 and 4. Changes in cardiac structure and function were examined by echocardiography. The  glucose and free fatty acids (FFA) were examined by ELISA. The expression of Notch, LKB1 and AMPK mRNA was examined by a real-time fluorescence quantitative method. The expression of glucose transporter 4 (GLUT4), and the expression of total acetyl-CoA carboxylase (ACC) and its phosphorylation were examined by western blotting.RESULTSCompared with the sham group, the expression of Notch, LKB1 and AMPK mRNA in the MI group was lower. Compared with the MI group, the expression of these mRNAs in the YQHX and perindopril groups was higher, and their expression in the DAPT group was lower. At all time points, the protein expression of GLUT4 and pACC decreased in the MI group. On week 1, the expression of pACC protein was higher. In the DAPT group, the expression of pACC protein decreased. Compared with the YQHX group, the expression of pACC protein in the DAPT + YQHX group was lower. On week 4, compared with the MI group, the expression of GLUT4 protein in the YQHX group and the perindopril group was higher. The expression of GLUT4 protein in the DAPT group decreased. Compared with the YQHX group, the expression of GLUT4 protein in the DAPT+YQHX group was lower. There was no significant difference in the expression of ACC protein between the groups.CONCLUSIONYQHX promoted cross talk between the LKB1-dependent Notch1 and AMPK in myocardial tissue after myocardial infarction. Furthermore, it regulated the glucose and lipid metabolism of cardiomyocytes at different time points, thereby ameliorating the cardiac energy metabolism via different mechanisms and protecting the heart.

Objective: To investigate how Yiqi Huoxue (YQHX) prescription regulates mitochondrial biosynthesis and ATP synthesis via AMP-activated protein kinase (AMPK) and to reveal its molecular mechanism in preventing and treating post-MI myocardial remodeling. Methods: The MI animal model of myocardial infarction were established by ligating Sprague Dawley (SD) rats' left anterior descending coronary arteries; the animals were randomly divided into MI group, YQHX prescription group and perindopril group, and a sham operation group was set at the same time. Related drug intervention was administered on the 2nd day after surgery, the YQHX group was given Astragalus, angelica, ginseng, Ligusticum wallichii and pseudoginseng, provided by the Dongzhimen Hospital of Beijing University of Chinese Medicine, once per day, at a dose of 21 g/kg body weight/day (the clinical equivalent dose based on a previous study), and the changes in relevant indicators were observed at 1 week and 4 weeks. Echocardiography (ECG) was used to observe the changes in rat cardiac structure and functions; the morphology-based technique was used to observe the changes in myocardial cells and mitochondria; the expression of AMPK signal pathway-related proteins and mRNA was detected using western blotting and real-time fluorescence quantification respectively, while fluorescence enzyme-labeled method was used for detecting ATP synthesis. Results: Cardiac structure and functions: Compared with the MI group at 1 week, the YQHX prescription group and the perindopril group exhibited increased LVEF and LVFS (Pall < .05); their LVEDs and LVEDd were reduced but suggested no statistically significant differences (F = 2.258, F = 0.3464, Pall > .05). At 4 weeks, both LVEF and LVFS were elevated in the YQHX prescription group (P = .008, .009) and the perindopril group (P = .279, .333), where differences in the later group indicated no statistical significance; the YQHX prescription group and the perindopril group were also featured by reduced LVEDs and LVEDd (Pall < .05). Morphology: Compared with the MI group at two time nodes, the YQHX prescription group and the perindopril group exhibited significant improvement in the pathological changes in myocardial cells and mitochondrial structure. Expression of AMPK signal pathway-related proteins and mRNA: The expression of both pLKB1 and pAMPK proteins followed a rising trend in the YQHX prescription group and the perindopril group at 1 week. The expression of LKB1mRNA and AMPKmRNA was elevated (Pall < .05). The increased expression of PGC-1α, NRF1 and mtTFA proteins demonstrated statistically significant differences (Pall < .05). The expression of mtDNA protein followed an increasing trend. At 4 weeks, the expression of both pLKB1 and pAMPK proteins was elevated (Pall < .05). Heightened expression was also reported in LKB1mRNA and AMPKmRNA (Pall < .05). The increased expression of PGC-1α, NRF1, mtTFA and mtDNA proteins demonstrated statistically significant differences (Pall < .05). ATP synthesis: ATP synthesis was increased in the YQHX prescription group and the perindopril group at both 1 week and 4 weeks (Pall < .001). Conclusion: The possible mechanism of YQHX prescription for preventing and treating post-MI myocardial remodeling may function through strengthening the activation AMPK signal pathway by LKB1, thus further increasing the expression of downstream transcription factor proteins and initiating mitochondrial replication and transcription; as a result, the YQHX prescription can improve the post-MI damage in mitochondrial morphological structure in the tissue at heart marginal zone as well as enhance mitochondrial biosynthesis and ATP synthesis.

Objective To investigate the regulatory mechanism of Tiaozhi Tongmai formula on vascular remodeling related factors in rabbits with atherosclerosis.Method The rabbit model of atherosclerosis was established by high fat diet and balloon injury.54 rabbits were randomly divided into normal group, model group, sham operation group, atorvastatin group and Chinese medicine group.Except the rabbits in normal group, the remaining four groups to choose the observation point at 45 day and 90 day after treatment.At 45 days, 30 animals were observed.At 90 days, 24 animals were observed.The lumen diameter and the intima-media thickness and the peak systolic velocity was measured by ultrasound.The common carotid arteries were observed by pathology.The levels of matrix metalloproteinases (MMP-1, MMP-2), tissue inhibitor of metalloproteinase-1(TIMP-1), platelet derived endothelial cell growth factor (PD-ECGF) and uric acid was detected by enzyme-linked immunosorbent assay(ELISA).Result There are two time points, the expression levels of MMP-1, MMP-2, PD-ECGF and UA in the model group were significantly higher than those in the sham operation group (P<0.01), TIMP-1 was lower than that in sham operation group(P<0.01).The serum levels of MMP-1, MMP-2, PD-ECGF and UA in the Tiaozhi Tongma formula group was significantly lower than those in the model group(P<0.01), the level of TIMP-1 was significantly increased(P<0.01), there was no significant difference from atorvastatin group.Conclusions Tiaozhi Tongmai formula can reduced the level of MMP-1, MMP-2,PD-ECGF, UA, and increase the level of TIMP-1, therefore influence the vascular remodeling to improve atherosclerosis, this may be the mechanism of its.

Objective:To conduct computing network pharmacological studies on Radix Astragali seu Hedysari,Radix Angelicae Sinensis,Radix Ginseng,Radix Notoginseng and Rhizoma Ligustici Chuanxiong of Yiqi Huoxue Formula,in order to explore the active constituents in the treatment of myocardial ischemia,clarify the effective constituents of this formula in treating myocardial ischemia,in order to provide a research support for clarifying the pharmacodynamic material basis and popularizing of this formula in future.Methods:Molecular docking and network analysis methods were applied to screen the active constituents of Yiqi Huoxue Formula in treating myocardial ischemia,in order to study the network pharmacological properties of molecules included in Yiqi Huoxue Formula in treating myocardial ischemia.Results:If the network degree was greater than 8,the betweenness and network degree between 20 compounds and 20 anti-myocardial ischemia targets were relatively high.Analysis results showed that Yiqi Huoxue Formula played a role in treating myocardial ischemia based on synergistic effect among the 20 effective constituents.Conclusion:Network pharmacology methods contributed to analyze the possible active molecules in Yiqi Huoxue Formula,which could treat myocardial ischemia disease by acting on the related targets.

Objective: Atherosclerosis is the fundamental pathophysiologic component of cardiovascular disease, and Tiaozhi Tongmai Granules show great efficiency in the treatment of the disease. However, the mechanism of Tiaozhi Tongmai Granules is still unclear. In this study, we have combined experiments with network pharmacology to explore the anti-atherosclerosis mechanism of Tiaozhi Tongmai Granules. Method: 120 male ApoE−/− mice were randomly divided into three groups: the model group, Chinese herb group and Atorvastatin group. The model group, Atorvastatin group and Chinese herb group were fed with a high-fat diet, a high-fat diet plus atorvastatin (5.1 mg/kg/d) and a high-fat diet plus Tiaozhi Tongmai Granules (16.5 g/kg/d) for 16 weeks, respectively. Atherogenesis was identified by H&E staining. The colocalization of neutral lipid stain BODIPY and microtubule-associated protein 1 light chain 3 (LC3) and the colocalization of BODIPY and lysosomal-associated membrane protein 1 (LAMP1) within ApoE−/− mice aortic plaques were tested using fluorescence confocal microscopy and the Pearson's coefficients were calculated. To further explore the anti-atherosclerosis mechanism of Tiaozhi Tongmai Granules, the network pharmacology was used to construct the herb-compound-target network. Results: The size of the aortic lipid plaque in the Chinese herb group and Atorvastatin group were smaller compared with the model group on the 16th week. Compared with the model group on the 16th week, the BODIPY and LC3 colocalization rate, the BODIPY and LAMP1 colocalization rate of the Chinese herb group and Atorvastatin group all presented significant increase in the aortic plaque (P < .001), showing that Tiaozhi Tongmai Granules could enhance autophagosome activity in the macrophage. In the herb-compound-target network, 17 active compounds and 27 targets were obtained through literature searching and using LHRI & DAVID Bioinformatics. It was found that 23 targets were correlated with the macrophage. Some of them participated in macrophage inflammatory response, and the other targets could promote/inhibit phagocytosis of the macrophage. It was hypothesized that the active compounds of Tiaozhi Tongmai Granules were acting on these targets and having y the biological effects. Conclusions: In the progression stage of atherosclerosis, Tiaozhi Tongmai Granules can still make the macrophage have higher autophagosome activity, and play a role of anti-atherogenesis.

Objective The purpose of this article is to through the animal experiment,study the effect of Yiqi-huoxue medicine on the emotional stress of myocardial ischemia model rats,adponectin contant in heart and brain,to provide a theoretical ideas and basis for Yiqi-huoxue medicine in prevention and treatment of cardiovascular diseases. Methods Adopt the method of intraperitoneal injection of epinephrine(5 mg/kg · d),and chronic unpredictability emotional stimuli to make complexsrats under emotional stress state. Animals are divided into five group:Blank group,ISO model group,Complex model group,Chinese medicine group,Western medicine groups. Using ultrasonic electrocardiogram,monitoring method to record the heart function,using the RT-PCR method for determinate the level of adponectin in heart and brain. According to these indexes of every group in the experimental process,respectively to observe contrast the changes of different groups after 14 and 28 days,treatment. Results After 32 days,the ejection fraction(EF)of complex model group,and fractional shortening(FS)were lower than those of blank group,the difference was statistically significant(P<0.01);adponectin contents in complex model group and ISO model group were higher than that in blank group,the difference was statistically significant(P<0.01 or P<0.05);adponectin contents in traditional Chinese medicine group and Western medicine group were lower than the complex model group,the difference was statistically significant(P<0.01 or P<0.05). Conclusion The reason why Yiqi-huoxue medicine can increase cardiac ejection fraction of rats in complex model group may be according to its effect on adponectin.