Pharmacological Mechanisms of Tinglizi against Chronic Heart Failure Determined by Network Pharmacology and Molecular Docking.

OBJECTIVE: Tinglizi has been extensively used to treat chronic heart failure (CHF) in modern times, but the material basis and pharmacological mechanisms are still unclear. To explore the material basis and corresponding potential targets and to elucidate the mechanism of Tinglizi, network pharmacology and molecular docking methods were utilized. METHODS: The main chemical compounds and potential targets of Tinglizi were collected from the pharmacological database analysis platform (TCMSP). The corresponding genes of related action targets were queried through gene cards and UniProt database. The corresponding genes of CHF-related targets were searched through Disgenet database, and the intersection targets were obtained by drawing Venn map with the target genes related to pharmacodynamic components. Then, drug targets and disease targets were intersected and put into STRING database to establish a protein interaction network. The "active ingredient-CHF target" network was constructed with Cytoscape 3.8.2. Finally, Gene Ontology (GO) Enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of intersection targets were analyzed using metascape. With the aid of SYBYL software, the key active ingredients and core targets were docked at molecular level, and the results were visualized by PyMOL software. Molecular docking was carried out to investigate interactions between active compounds and potential targets. RESULTS: A total of 12 active components in Tinglizi were chosen from the TCMSP database, and 193 corresponding targets were predicted. Twenty-nine potential targets of Tinglizi on CHF were obtained, of which nine were the core targets of this study. Twenty GO items were obtained by GO function enrichment analysis (P < 0.05), and 10 signal pathways were screened by KEGG pathway enrichment analysis (P < 0.05), which is closely related to the treatment of CHF by Tinglizi. The constructed drug compound composition action target disease network shows that quercetin, kaempferol, and other active compounds play a key role in the whole network. The results of molecular docking showed that all the key active ingredients, such as quercetin and isorhamnetin, were able to successfully dock with ADRB2 and HMOX1 with a total score above 5.0, suggesting that these key components have a strong binding force with the targets. CONCLUSION: Through network pharmacology and molecular docking technology, we found that the main components of Tinglizi in the treatment of CHF are quercetin, kaempferol, ß-sitosterol, isorhamnetin, and so on. The action targets are beta 2-adrenergic receptor (ADRB2), heme oxygenase 1 (HMOX1), and so on. The main pathways are advanced glycation end products/receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications, hypoxia-inducible factor (HIF-1) signaling pathway, estrogen signaling pathway, and so on. They play an integrated role in the treatment of CHF.

Study on the Differences between Traditional Chinese Medicine Syndromes in NYHA I-IV Classification of Chronic Heart Failure.

OBJECTIVES: This study investigated the distribution of characteristics of traditional Chinese medicine syndromes and their association with symptoms in 1027 patients with chronic heart failure (CHF). METHODS: An observational study was performed by researchers, collecting data from 1036 patients with CHF from 24 Chinese medicine hospitals from May 2009 to December 2014. Due to incomplete information from nine patients, 1027 patients with CHF were analysed. The distribution of syndromes in CHF and association between high-frequency syndromes and symptoms were investigated. RESULTS: The primary syndromes were qi deficiency, blood stasis, fluid retention, yin deficiency, phlegm turbidity, and yang deficiency. The primary sites of disease were the heart, kidney, lung, and spleen. In patients with CHF of differing cardiac function, there was no significant difference in the frequency of yin deficiency (P>0.05). The distribution of yang deficiency was significantly different between New York Heat Association (NYHA) classes II, III, and IV and between classes I+II and III+IV (P<0.05). The frequency of phlegm turbidity was significantly different between NYHA classes II and III, between classes III and IV, and between classes I+II and III+IV (P<0.05). The frequency of fluid retention was significantly different between NYHA classes I and IV, between classes II, III, and IV, and between classes I+II and III+IV (P<0.05). Regarding associations between syndromes and symptoms, qi deficiency was diagnosed in 87.43% of patients with insomnia and spiritlessness; blood stasis in 84.85% of patients with spontaneous sweating + cyanosis of the lips; fluid retention in 75% of patients with a hard pulse and oedema; and yin deficiency in 72.92% of patients with feverish sensation in the chest, palms, and soles and spontaneous sweating. CONCLUSIONS: The frequency of yang deficiency and fluid retention was higher and that of phlegm turbidity was lower in classes III and IV than in classes I and II.

Identification of the potential crucial genes in invasive ductal carcinoma using bioinformatics analysis.

Invasive ductal carcinoma (IDC) is a common histological type of breast cancer. The aim of this study was to identify the potential crucial genes associated with IDC and to provide valid biological information for further investigations. The gene expression profiles of GSE10780 which contained 42 histologically normal breast tissues and 143 IDC tissues were downloaded from the GEO database. Functional and pathway enrichment analysis of differentially expressed genes (DEGs) were performed and protein-protein interaction (PPI) network was analyzed using Cytoscape. In total, 999 DEGs were identified, including 667 up-regulated and 332 down-regulated DEGs. Gene ontology analysis demonstrated that most DEGs were significantly enriched in mitotic cell cycle, adhesion and protein binding process. Through PPI network analysis, a significant module was screened out, and the top 10 hub genes, CDK1, CCNB1, CENPE, CENPA, PLK1, CDC20, MAD2L1, HIST1H2BK, KIF2C and CCNA2 were identified from the PPI network. The expression levels of the 10 genes were validated in Oncomine database. KIF2C, MAD2L1 and PLK1 were associated with the overall survival. And we used cBioPortal to explore the genetic alterations of hub genes and potential drugs. In conclusion, the present study identified DEGs between normal and IDC samples, which could improve our understanding of the molecular mechanisms in the development of IDC, and these candidate genes might be used as therapeutic targets for IDC.

Exosomes from high glucose-treated glomerular endothelial cells trigger the epithelial-mesenchymal transition and dysfunction of podocytes.

New data indicate that abnormal glomerular endothelial cell (GEC)-podocyte crosstalk plays a critical role in diabetic nephropathy (DN). The aim of our study is to investigate the role of exosomes from high glucose (HG)-treated GECs in the epithelial-mesenchymal transition (EMT) and dysfunction of podocytes. In this study, exosomes were extracted from GEC culture supernatants and podocytes were incubated with the GEC-derived exosomes. Here, we demonstrate that HG induces the endothelial-mesenchymal transition (EndoMT) of GECs and HG-treated cells undergoing the EndoMT secrete more exosomes than normal glucose (NG)-treated GECs. We show that GEC-derived exosomes can be internalized by podocytes and exosomes from HG-treated cells undergoing an EndoMT-like process can trigger the podocyte EMT and barrier dysfunction. Our study reveals that TGF-ß1 mRNA is enriched in exosomes from HG-treated GECs and probably mediates the EMT and dysfunction of podocytes. In addition, our experimental results illustrate that canonical Wnt/ß-catenin signaling is involved in the exosome-induced podocyte EMT. Our findings suggest the importance of paracrine communication via exosomes between cells undergoing the EndoMT and podocytes for renal fibrosis in DN. Thus, protecting GECs from the EndoMT and inhibiting TGF-ß1-containing exosomes release from GECs is necessary to manage renal fibrosis in DN.

Tongxinluo Inhibits Renal Fibrosis in Diabetic Nephropathy: Involvement of the Suppression of Intercellular Transfer of TGF-[Formula: see text]1-Containing Exosomes from GECs to GMCs.

Glomerular mesangial cells (GMCs) activation is implicated in the pathogenesis of diabetic nephropathy (DN). Our previous study revealed that high glucose (HG)-treated glomerular endothelial cells (GECs) produce an increased number of TGF-[Formula: see text]1-containing exosomes to activate GMCs through the TGF-[Formula: see text]1/Smad3 signaling pathway. We also identified that Tongxinluo (TXL), a traditional Chinese medicine, has beneficial effects on the treatment of DN in DN patients and type 2 diabetic mice. However, it remained elusive whether TXL could ameliorate renal structure and function through suppression of intercellular transfer of TGF-[Formula: see text]1-containing exosomes from GECs to GMCs. In this study, we demonstrate that TXL can inhibit the secretion of TGF-[Formula: see text]1-containing exosomes from HG-treated GECs. Furthermore, exosomes produced by HG induced-GECs treated with TXL cannot trigger GMC activation, proliferation and extracellular matrix (ECM) overproduction both in vitro and in vivo. These results suggest that TXL can prevent the transfer of TGF-[Formula: see text]1 from GECs to GMCs via exosomes, which may be one of the mechanisms of TXL in the treatment of DN.

Chinese Herbal Medicine in the Treatment of Chronic Heart Failure: Three-Stage Study Protocol for a Randomized Controlled Trial.

Background. Chinese herbal medicine (CHM) has been used in the treatment of chronic heart failure (CHF) for a long time. Treatment based on syndrome differentiation and the main characteristic of TCM is the fundamental principle of TCM practice. In this study protocol, we have designed a trial to assess the efficacy and safety of CHM on CHF based on syndrome differentiation. Methods/Design. This is a three-stage trial of CHM in the treatment of CHF. The first stage is a literature review aiming to explore the common syndromes of CHF. The second is a multicentral, randomized, placebo-controlled trial to evaluate the efficacy and safety of CHM for the treatment of CHF. The third is a multicentral, randomized controlled clinical trial aiming to make cost-effectiveness analysis and evaluate the feasibility, compliance, and universality of CHM on CHF. Discussion. This trial will evaluate the efficacy, safety, feasibility, compliance, and universality of CHM on CHF. The expected outcome is to provide evidence-based recommendations for CHM on CHF and develop a prescription of CHM in the treatment of CHF. This trial is registered with NCT01939236 (Stage Two of the whole trial).

[A correlation research on Chinese medical syndromes of chronic heart failure and various complications].

OBJECTIVE: To study the correlation between various complications of chronic heart failure (CHF) patients and Chinese medical syndromes, thus indicating distribution laws of Chinese medical syndromes in various complications of CHF patients. METHODS: Chinese medical syndrome typing was performed in 630 CHF patients by cross-sectional study of the demographic data, history of present diseases, related information on Chinese medical four diagnostic methods, and the distribution of complications. Logistic regression analysis was used to determine the correlation of various complications of CHF patients and Chinese medical syndromes. RESULTS: In this study, recruited were common complications such as hypertension, diabetes, arrhythmia, hyperlipemia, and cerebral vascular accident, and so on. Main syndromes were sequenced as qi deficiency syndrome, blood stasis syndrome, water retention syndrome, yin deficiency syndrome, phlegm turbid syndrome, yang deficiency syndrome. Results of Logistic regression analysis indicated that correlation existed between common complications and Chinese medical syndromes. In CHF complicated hypertension patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with hypertension (P < 0.05). In CHF complicated diabetes patients, Logistic regression analysis showed phlegm turbid syndrome and water retention syndrome were positively correlated with diabetes (P < 0.05). In CHF complicated arrhythmia patients, there was no statistical difference in the distribution of each syndrome (P > 0.05). In CHF complicated hyperlipemia patients, Logistic regression analysis showed qi deficiency syndrome and water retention syndrome were negatively correlated with hyperlipemia (P < 0.05), while blood stasis syndrome, yin deficiency syndrome, and phlegm turbid syndrome were positively correlated with hyperlipemia (P < 0.01). In CHF complicated cerebral vascular accident patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with cerebral vascular accident (P < 0.01, P < 0.05). CONCLUSIONS: There existed certain correlations between complications of CHF and the distribution of main Chinese medical syndromes. It could be used as guidance for treating CHF and its various complications by Chinese medicine and pharmacy.

A New Biomarkers Feature Pattern Consisting of TNF- α , IL-10, and IL-8 for Blood Stasis Syndrome with Myocardial Ischemia.

Objective. To explore new diagnostic patterns for syndromes to overcome the insufficiency of obtainable macrocharacteristics and specific biomarkers. Methods. Chinese miniswines were subjected to Ameroid constrictor, placed around the proximal left anterior descending branch. On the 4th week, macrocharacteristics, coronary angiography, echocardiography, and hemorheology indices were detected for diagnosis. IL-1, IL-6, IL-8, IL-10, TNF- α , and hsCRP in serum were detected, and Decision Tree was built. Results. According to current official-issued standard, model animals matched the diagnosis of blood stasis syndrome with myocardial ischemia based on findings, including >90% occlusion, attenuated left ventricular segmental motion, dark red or purple tongues, and higher blood viscosity. Significant decrease of IL-10 and increase of TNF- α were found in model animals. However, in the Decision Tree, besides IL-10 and TNF- α , IL-8 helped to increase the accuracy of classification to 86%. Conclusions. The Decision Tree building with TNF- α , IL-10, and IL-8 is helpful for the diagnosis of blood stasis syndrome in myocardial ischemia animals. What is more is that our data set up a new path to the differentiation of syndrome by feature patterns consisting of multiple biomarkers not only for animals but also for patients. We believe that it will contribute to the standardization and international application of syndromes.

Effects of traditional Chinese medicine on rats with Type II diabetes induced by high-fat diet and streptozotocin: a urine metabonomic study.

BACKGROUND: Type II diabetes has become a serious threat to human health in recent years. Among adults above 20 years old in China, the prevalence rate of diabetes is 9.7%. Thus, it is imperative to study the mechanisms underlying type II diabetes to develop effective therapeutic treatments. OBJECTIVE: To examine metabolic changes in a rat model of type II diabetes and explore mechanisms underlying the beneficial effects of traditional Chinese medicine (TCM) in this model. 120 rats were divided into four groups, including a control group, a high-fat diet group (high-fat diet and streptozotocin injection), a TCM group (high-fat diet, streptozotocin injection, followed by TCM administration), and a rosiglitazone maleate group (high-fat diet, streptozotocin injection, followed by rosiglitazone maleate administration). Metabolites in urine samples from 1-3 weeks (time point 1) and 4-6 weeks (time point 2) of drug administration were compared by gas chromatography-mass spectrometry (GC-MS). RESULTS: Our results showed that in the high-fat diet group, at time point 2, the levels of dihydroxybenzoic acid, L-ascorbic acid, D-gluconic acid, octadecanoic acid, and glutaric acid in urine were significantly higher than at time point 1. In the TCM group, at time point 2, the urine levels of L-ascorbic acid were markedly lower than at time point 1. CONCLUSION: Our studies demonstrated that examining urine metabolic changes provided important insights into the mechanisms underlying type II diabetes as well as the therapeutic effects of TCM.

Metabolomic identification of diagnostic plasma biomarkers in humans with chronic heart failure.

Chronic heart failure (CHF), as a progressive clinical syndrome, is characterized by failure of enough blood supply from the heart to meet the body's metabolic demands, and there is intense interest in identifying novel biomarkers that could make contributions to the diagnosis of CHF. Metabolomics, compared with current diagnostic approaches, could investigate many metabolic perturbations within biological systems. The overarching goal of the work discussed here is to apply a high-throughput approach to identify metabolic signatures and plasma diagnostic biomarkers underlying CHF by 1H-NMR spectroscopy. Plasma samples from 39 patients with CHF and 15 controls were analyzed by NMR spectroscopy. After processing the data, orthogonal partial least square discriminant analysis (OPLS-DA) was performed. The statistical model revealed good explained variance and predictability, and the diagnostic performance assessed by leave-one-out analysis exhibited 92.31% sensitivity and 86.67% specificity. The OPLS-DA score plots of spectra revealed good separation between case and control on the level of metabolites, and multiple biochemical changes indicated hyperlipidemia, alteration of energy metabolism and other potential biological mechanisms underlying CHF. It was concluded that the NMR-based metabolomics approach demonstrated good performance to identify diagnostic plasma markers and provided new insights into metabolic process related to CHF.

Clinical data mining of phenotypic network in angina pectoris of coronary heart disease.

Coronary heart disease (CHD) is the leading causes of morbidity and mortality in China. The diagnosis of CHD in Traditional Chinese Medicine (TCM) was mainly based on experience in the past. In this paper, we proposed four MI-based association algorithms to analyze phenotype networks of CHD, and established scale of syndromes to automatically generate the diagnosis of patients based on their phenotypes. We also compared the change of core syndromes that CHD were combined with other diseases, and presented the different phenotype spectra.

The active ingredients of Jiang-Zhi-Ning: study of the Nelumbo nucifera alkaloids and their main bioactive metabolites.

The object of this study was to identify the major active ingredients of the Chinese Traditional Medicine Jiang-Zhi-Ning (JZN) based on the high performance liquid chromatography (HPLC) profiles of plasma samples obtained from beagle dogs at different times after intragastric administration of JZN, crude JZN extracts, different extracted fractions, different subfractions of the active fraction and different isolated ingredients. 2-Hydroxy-1-methoxyaporphin (2H1M), an alkaloid from Nelumbo nucifera, one of the herbs that make up JZN, was identified as the constituent showing the major pharmacodynamic effect. The major metabolites of 2H1M were analyzed and identified as N-demethyl-2-hydroxy-1-methoxyaporphine-2-O-glycuronic acid, 2-hydroxy-1-methoxy-aporphine-2-O-glycuronic acid and 2-hydroxy-1-methoxy-aporphine-2-O-sulphate. This study provided a comprehensive insight into the active components of JZN.

Metabolomics-based study of clinical and animal plasma samples in coronary heart disease with blood stasis syndrome.

The aim of this study is to explore a bridge connecting the mechanism basis and macro syndromes of coronary heart disease with experimental animal models. GC-MS technique was used to detect the metabolites of plasma samples in mini swine models with myocardial infarction (MI) and patients with unstable angina (UA). 30 metabolites were detected in the plasma samples of more than 50 percent of model group and control group in swine, while 37 metabolites were found in the plasma samples of UA patients and healthy control group. 21 metabolites in the plasma samples of swine model and 20 metabolites in patients with UA were found of significant value. Among which, 8 shared metabolites were found of low level expression in both swine model and UA patients. Independent Student's t-test, principal component analysis (PCA), and hierarchicalcluster analysis (HCA) were orderly applied to comprehend inner rules of variables in the data. The 8 shared metabolites could take place of the 21 or 20 metabolites in classification of swine model with MI and UA patients, which could be considered as a bridge connecting the mechanism basis and macrosyndromes of swine model with MI and UA patients.

The effects of jiang-zhi-ning and its main components on cholesterol metabolism.

To examine how Jiang-Zhi-Ning (JZN) regulates cholesterol metabolism and compare the role of its four main components. We established a beagle model of hyperlipidemia, fed with JZN extract and collected JZN-containing serum 0, 1, 2, 4, and 6 h later. Human liver cells Bel-7402 were stimulated with 10% JZN-containing serum as well as the four main components of JZN and Atorvastatin. The mRNA expression of LDL receptor (LDL-R), 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoAR), cytochrome P450 7A1 (CYP7A1), and acetyl-Coenzyme A acetyltransferase 2 (ACAT2) was measured by real-time PCR. LDL-R surface expression and LDL-binding and internalization were examined by flow cytometry. The results showed that JZN-containing serum significantly increased the mRNA expression of LDL-R, HMG-CoAR, and CYP7A1 in Bel-7402 cells. All the four components significantly increased the mRNA and protein expression of LDL-R and HMG-CoAR and decreased the mRNA and protein expression of ACAT2 in Bel-7402 cells. Hyperinand chrysophanol also markedly increased the mRNA expression of CYP7A1. Stimulation with stilbene glycosidesignificantly increased the surface expression of LDL-R and the binding and internalization of LDL. In conclusion, JZN and its four components have close relationship with the process of cholesterol metabolism, emphasizing their promising application as new drug candidates in the treatment of hyperlipidemia.

In silico syndrome prediction for coronary artery disease in traditional chinese medicine.

Coronary artery disease (CAD) is the leading causes of deaths in the world. The differentiation of syndrome (ZHENG) is the criterion of diagnosis and therapeutic in TCM. Therefore, syndrome prediction in silico can be improving the performance of treatment. In this paper, we present a Bayesian network framework to construct a high-confidence syndrome predictor based on the optimum subset, that is, collected by Support Vector Machine (SVM) feature selection. Syndrome of CAD can be divided into asthenia and sthenia syndromes. According to the hierarchical characteristics of syndrome, we firstly label every case three types of syndrome (asthenia, sthenia, or both) to solve several syndromes with some patients. On basis of the three syndromes' classes, we design SVM feature selection to achieve the optimum symptom subset and compare this subset with Markov blanket feature select using ROC. Using this subset, the six predictors of CAD's syndrome are constructed by the Bayesian network technique. We also design Naïve Bayes, C4.5 Logistic, Radial basis function (RBF) network compared with Bayesian network. In a conclusion, the Bayesian network method based on the optimum symptoms shows a practical method to predict six syndromes of CAD in TCM.