Well-known Traditional Chinese Medicine Formula Research Articles

Exploration on pharmacological mechanisms of YZP against neuropathic pain via inhibiting spinal inflammation and the rationality of its compatibility

Ethnopharmacological relevanceYuanhu Zhitong Prescription (YZP) is a well-known traditional Chinese medicine (TCM) formula for neuropathic pain (NP) therapy with a satisfying clinical efficacy. However, the underlying pharmacological mechanism and its compatibility principle remain unclear. Aim of the studyThis study aims to investigate the analgesic and compatibility mechanisms of YZP on neuropathic pain (NP) at the gene and biological process levels. Materials and methodsThe chronic constriction injury (CCI) rats were intragastrically administrated with extracts of YZP, YH and BZ separately, and then mechanical hypersensitivity were measured to evaluate the analgesic effects between YH and BZ before and after compatibility. Then, RNA-seq and bioinformatics analyses were performed to elucidate the potential mechanisms underlying YZP's analgesia and compatibility. Finally, the expression levels and significant differences of key genes were analyzed. ResultsBehaviorally, both YZP and YH effectively alleviated mechanical allodynia in CCI rats, with YZP being superior to YH. In contrast, we did not observe an analgesic effect of BZ. Genetically, YZP, YH, and BZ reversed the expression levels of 52, 34, and 42 aberrant genes in the spinal cord of CCI rats, respectively. Mechanically, YZP was revealed to alleviate NP mainly by modulating the inflammatory response and neuropeptide signaling pathway, which are the dominant effective processes of YH. Interestingly, the effective targets of YZP were especially enriched in leukocyte activation and cytokine-mediated signaling pathways. Moreover, BZ was found to exert an adjunctive effect in enhancing the analgesic effect of YH by promoting skeletal muscle tissue regeneration and modulating calcium ion transport. ConclusionsYH, as the monarch drug, plays a dominant role in the analgesic effect of YZP that effectively relieves NP by inhibiting the spinal inflammation and neuropeptide signaling pathway. BZ, as the minister drug, not only synergistically enhances analgesic processes of YH but also helps to alleviate the accompanying symptoms of NP. Consequently, YZP exerted a more potent analgesic effect than YH and BZ alone. In conclusion, our findings offer new insights into understanding the pharmacological mechanism and compatibility principle of YZP, which may support its clinical application in NP therapy.

Toxicity studies of condensed fuzheng extract in mice and rats

Nutritional supplements have been used to improve immune function. Condensed fuzheng extract (CFE) is a well-known traditional Chinese medicine (TCM) formula that is predominantly made from sheep placenta, Astragalus mongholicus Bunge, and Polygonatum kingianum Collett & Hemsl. However, the toxicological profile of CFE has not been determined. In this study, we investigated the acute (14 days) and sub-chronic (90 days) oral toxicities of CFE in mice and rats and the phytochemical composition of CFE. Materials and methodsFor the assessment of acute toxicity, 80 ICR mice of both sexes were randomly divided into four groups. Three groups were treated with 4500, 2250 and 1125 mg/kg/d bw CFE daily (n = 10/group per sex) for 14 days; a separate group was used as control. To test the sub-chronic toxicity, male and female Sprague Dawley rats were orally administered 8150, 4075 or 2037 mg/kg bw of CFE for 90 days; a control group was included. Hematological, biochemical, and histopathological markers were tested at the end of the experiment. The chemical composition of CFE was determined by UPLC-HRMS method. ResultsIn both acute and sub-chronic toxicity studies, no mortalities, indications of abnormality, or treatment-related adverse effects were observed. The LD50 of CFE was higher than 4500 mg/kg. There were no significant changes in the hematological and biochemical data in the treatment group compared with the control group (p > 0.05). Histopathological analyses of the heart, liver, spleen, lungs, kidneys, thymus, testes (male rats) and ovaries (female rats) revealed no anatomical changes of each organ. Phytochemical analysis of CFE revealed the presence of flavonoids (highest abundance), phenols and alkaloids. In conclusion, our results showed that CFE is a safe and non-toxic formula. We also reported phytochemicals in CFE that may possess important pharmacological effects.

Computer-aided investigation of Traditional Chinese Medicine mechanisms: A case study of San-Ao decoction in asthma treatment

The San-Ao Decoction (SAD) is a well-known Traditional Chinese Medicine (TCM) formula used to alleviate respiratory symptoms, including asthma. However, its precise mechanisms of action have remained largely unknown. In this study, we utilized computer-aided approaches to explore these mechanisms. Firstly, we conducted a comprehensive analysis of the chemical composition of SAD, which allowed us to identify the 28 main ingredients. Then, we employed computer simulations to investigate the potential active ingredients of SAD and the corresponding binding sites of transient receptor potential vanilloid 1 (TRPV1). The simulations revealed that D509 and D647 were the potential binding sites for TRPV1. Notably, molecular dynamics (MD) studies indicated that site D509 may function as an allosteric site of TRPV1. Furthermore, to validate the computer-aided predictions, we performed experimental studies, including in vitro and in vivo assays. The results of these experiments confirmed the predictions made by our computational models, providing further evidence for the mechanisms of action of San-Ao Decoction in asthma treatment. Our findings demonstrated that: i) D509 and D647 of TRPV1 are the key binding sites for the main ingredients of SAD; ii) SAD or its main ingredients significantly reduce the influx of Ca2+ through TRPV1, following the TCM principle of “Jun, Chen, Zuo, Shi”; iii) SAD shows efficiency in comprehensive in vivo validation. In conclusion, our computer-aided investigation of San-Ao Decoction in asthma treatment has provided valuable insights into the therapeutic mechanisms of this TCM formula. The combination of computational analysis and experimental validation has proven effective in enhancing our understanding of TCM and may pave the way for future discoveries in the field.

Integration of non-targeted multicomponent profiling, targeted characteristic chromatograms and quantitative to accomplish systematic quality evaluation strategy of Huo-Xiang-Zheng-Qi oral liquid

Huo-Xiang-Zheng-Qi oral liquid (HXZQOL) is a well-known traditional Chinese medicine formula for the treatment of gastrointestinal diseases, with the pharmacologic effects of antiinflammatory, immune protection and gastrointestinal motility regulation. More significantly, HXZQOL is recommended for the treatment of COVID-19 patients with gastrointestinal symptoms, and it has been clinically proven to reduce the inflammatory response in patients with COVID-19. However, the effective and overall quality control of HXZQOL is currently limited due to its complex composition, especially the large amount of volatile and non-volatile active components involved. In this study, aimed to fully develop a comprehensive strategy based on non-targeted multicomponent identification, targeted authentication and quantitative analysis for quality evaluation of HXZQOL from different batches. Firstly, the non-targeted high-definition MSE (HDMSE) approach is established based on UHPLC/IM-QTOF-MS, utilized for multicomponent comprehensive characterization of HXZQOL. Combined with in house library-driven automated peak annotation and comparison of 47 reference compounds, 195 components were initially identified. In addition, HS-SPME-GC-MS was employed to analyze the volatile organic compounds (VOCs) in HXZQOL, and a total of 61 components were identified by comparison to the NIST database, reference compounds as well as retention indices. Secondly, based on the selective ion monitoring (SIM) of 24 “identity markers” (involving each herbal medicine), characteristic chromatograms (CCs) were established on LC-MS and GC-MS respectively, to authenticate 15 batches of HXZQOL samples. The targeted-SIM CCs showed that all marker compounds in 15 batches of samples could be accurately monitored, which could indicate preparations authenticity. Finally, a parallel reaction monitoring (PRM) method was established and validated to quantify the nine compounds in 15 batches of HXZQOL. Conclusively, this study first reports chemical-material basis, SIM CCs and quality evaluation of HXZQOL, which is of great implication to quality control and ensuring the authenticity of the preparation.

Shengxian decoction protects against chronic heart failure in a rat model via energy regulation mechanisms

BackgroundChronic heart failure (CHF) is actually a disease caused by an imbalanced energy metabolism between myocardial energy demand and supply, ultimately resulting in abnormal myocardial cell structure and function. Energy metabolism imbalance plays an important role in the pathological process of chronic heart failure (CHF). Improving myocardial energy metabolism is a new strategy for the treatment of CHF. Shengxian decoction (SXT), a well-known traditional Chinese medicine (TCM) formula, has good therapeutic effects on the cardiovascular system. However, the effects of SXT on the energy metabolism of CHF is unclear. In this study, we probed the regulating effects of SXT on energy metabolism in CHF rats using various research methods.MethodsHigh-performance liquid chromatography (HPLC) analysis was used to perform quality control of SXT preparations. Then, SD rats were randomly assigned into 6 groups: sham, model, positive control (trimetazidine) and high-, middle-, and low-dose SXT groups. Specific reagent kits were used to detect the expression levels of ALT and AST in rats’ serum. Echocardiography was used to evaluate cardiac function. H&E, Masson and TUNEL staining were performed to examine myocardial structure and myocardial apoptosis. Colorimetry was used to determine myocardial ATP levels in experimental rats. Transmission electron microscopy was used to observe the ultrastructure of myocardial mitochondria. ELISA was used to estimate CK, cTnI, and NT-proBNP levels, and LA、FFA、MDA、SOD levels. Finally, Western blotting was used to examine the protein expression of CPT-1, GLUT4, AMPK, p-AMPK, PGC-1α, NRF1, mtTFA and ATP5D in the myocardium.ResultsHPLC showed that our SXT preparation method was feasible. The results of ALT and AST tests indicate that SXT has no side effect on the liver function of rats. Treatment with SXT improved cardiac function and ventricular remodelling and inhibited cardiomyocyte apoptosis and oxidative stress levels induced by CHF. Moreover, CHF caused decrease ATP synthesis, which was accompanied by a reduction in ATP 5D protein levels, damage to mitochondrial structure, abnormal glucose and lipid metabolism, and changes in the expression of PGC-1α related signal pathway proteins, all of which were significantly alleviated by treatment with SXT.ConclusionSXT reverses CHF-induced cardiac dysfunction and maintains the integrity of myocardial structure by regulating energy metabolism. The beneficial effect of SXT on energy metabolism may be related to regulating the expression of the PGC-1α signalling pathway.

Shengjiang San alleviated sepsis-induced lung injury through its bidirectional regulatory effect

BackgroundSepsis is a life-threatening organ dysfunction caused by dysregulated host responses to infection, for which effective therapeutic strategies are still absent. Shengjiang San (SJS), a well-known Traditional Chinese Medicine formula, has been widely used clinically. However, its role in sepsis-induced lung injury remains unclear.MethodsTo explore its specific mechanism, we firstly established a sepsis animal model using cecal ligation and puncture (CLP) and treated MH-S cells with LPS plus ATP. Then, UPLC/Q-TOF–MS/MS was utilized to identify its active ingredients. Network pharmacology analysis was performed to uncover the potential mechanism. HE staining and biochemical analysis were conducted to validate its therapeutic effect. ELISA was applied to detect the release of pro-inflammatory and anti-inflammatory cytokines. Western blot was utilized to detect the protein levels of GSDMD, NLRP3, P65, ASC and caspase-1.ResultsSJS could dramatically increase the survival rate of sepsis. In addition, it is able to inhibit the pro-inflammatory cytokines release at day 1 post CLP while promote their production at day 7, indicating SJS could attenuate uncontrolled inflammatory response in the early stage and improve immunosuppression in the late phase. Network pharmacology analysis showed that pyroptosis is the crucial action SJS exerted in the protection of sepsis-induced lung injury. Western blot data implicated SJS could attenuate pyroptosis in early sepsis while enhance in the late phase.ConclusionsSJS acted to alleviate sepsis-induced lung injury through its bidirectional regulatory effect.

Comprehensive multicomponent characterization and quality assessment of Shuang-Huang-Lian powder injection using ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry and ultra-high-performance liquid chromatography-quadrupole-Orbitrap-mass spectrometry.

Shuang-Huang-Lian powder injection (SHLPI) is a well-known modern traditional Chinese medicine formula preparation (TCMFP) widely used to treat acute upper respiratory infections. However, SHLPI is extracted from pure Chinese medicine and administered through an injection, and many adverse reactions have been reported clinically. Therefore, it is necessary to characterize in depth the chemical composition of SHLPI and quantitatively analyze its potential allergenic components. In this study, the samples were analyzed using ion mobility ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UHPLC-QTOF-MS) combined with a self-built database. Furthermore, the parallel reaction monitoring (PRM) model of ultra-high-performance liquid chromatography-quadrupole-Orbitrap-mass spectrometry (UHPLC-Q-Orbitrap-MS) was used to successfully quantify 10 representative bioactive components. Using this strategy 90 compounds were identified, the fragmentation pathways of five representative compounds in the five main components of SHLPI were summarized, and 10 components (neochlorogenic acid, chlorogenic acid, sweroside, forsythiaside A, luteoloside, isochlorogenic acid B, isochlorogenic acid C, baicalin, phillyrin, and baicalein) were determine as the quality markers of SHLPI based on UPLC-Q-Orbitrap-MS. This work comprehensively characterized the material basis of SHLPI, summarized the cracking laws of representative substances, and quantitatively analyzed 10 potential allergenic components. Therefore, this study could provide a basis for the quality control of SHLPI and the clinical rational use of drugs to reduce its adverse reactions.

Multilevel data integration and molecular docking approach to systematically elucidate the underlying pharmacological mechanisms of Er-Zhi-Wan against hepatocellular carcinoma.

As a multicomponent, multitarget empirical therapy, traditional Chinese medicine (TCM) has been used clinically in Asia for thousands of years. Due to this unique feature, TCM therapy is considered a promising therapeutic strategy for the treatment of hepatocellular carcinoma (HCC). Er-Zhi-Wan (EZW), a well-known TCM formula containing two herbs, Fructus Ligustri Lucidi (FLL, Nü-Zhen-Zi) and Ecliptae Herba (EH, Mo-Han-Lian), is commonly used in clinical practice to prevent and treat liver diseases. Modern pharmacological studies have shown that both EH and FLL can inhibit HCC proliferation. However, the pharmacological mechanism, potential targets, and clinical value of EZW in inhibiting HCC have not been fully elucidated. We used multilevel databases (Gene Expression Omnibus (GEO), Traditional Chinese Medicine Systems Pharmacology (TCMSP), High-throughput Experiment- and Reference-guided database (HERB), and SwissTargetPrediction) to show that EZW suppresses HCC through 19 active components acting on 66 potential targets. Enrichment analysis revealed that EZW mainly regulates HCC progression through various metabolic pathways, the cell cycle, and cellular senescence. Furthermore, we used The Cancer Genome Atlas (TCGA)-LIHC database to analyze the expression patterns and clinical characteristics of cellular senescence-related genes and identified CDK1, CDK4, CHEK1, and G6PD as key therapeutic molecular targets in EZW-suppressed HCC. Molecular docking revealed that EZW could exert its anti-HCC effect by binding various active components to the above cellular senescence-related genes and regulating their activities. In conclusion, we systematically revealed the potential pharmacological mechanisms and molecular targets of EZW against HCC based on multilevel data integration and a molecular docking strategy.

QingreHuoxue decoction protects joint and toe bone morphology and structure in rats with active RA through bidirectional regulation of bone metabolism

QingreHuoxue decoction (QRHX) is a well-known traditional Chinese medicine formula for treating rheumatoid arthritis (RA). It is not known how QRHX is involved in bone metabolism regulation and thereby alleviatesbone injury caused by arthritis. In this study, we used collagen-induced arthritis (CIA) rat model to evaluate the effects of QRHX onarthriticdiarthrodial joints and the underlying molecular mechanisms. Our results showed that QRHX significantly reduced foot joint volume, synovial inflammatory infiltration and the arthritis index (AI) score. Micro-CT imaging showed that toe bone and ankle joint were eroded and destroyed progressively during RA process, andcompensatory hyperplasia appeared in late stage RA. QRHX treatment inhibited foot bone erosion of early and mid-stage RA, and improved bone mineral density and trabecular fine structure in late RA.This studyunlocked the whole process details of ankle and toe bone injuryduring RA, as well as the bone metabolic characteristics under QRHX treatmentin CIArats. QRHX was confirmed to inhibite the expression of RANKL and DKK-1 mRNA and protein, enhance the expression of β-catenin. Meanwhile, QRHX reduced the pro-inflammatorycytokines IL-17, IL-1β and TNF-αand upregulated anti-inflammatory IL-4 in serum, improving local and systemic inflammatory microenvironment. These results demonstrated that QRHX had sustained protective effects on diarthrodial joints/bone during RA by a synergistic regulation mechanism, which reverses the imbalance of bone metabolismthroughinhibiting RANKL related osteoclast pathway and promotingWnt/β-catenin related osteoblast activation pathway.

A multidimensional strategy to rapidly identify the chemical constituents in Shengxian Decoction by using ultra-performance liquid chromatography coupled with ion mobility spectrometry quadrupole time-of-flight mass spectrometry.

As a well-known traditional Chinese medicine formula, the chemical constituents of Shengxian Decoction still remain unclear due to its complexity. In this study, a multidimensional strategy based on ultra-performance liquid chromatography coupled with ion mobility spectrometry quadrupole time-of-flight mass spectrometry and informatics UNIFI platform was applied to achieve rapid and comprehensive identification of the complex composition of Shengxian Decoction. Data-independent acquisition, fast data-directed analysis, and high-definition MSE were used to obtain more and cleaner mass spectrum information. As a result, a total of 120 compounds including 74 saponins, 17 flavonoids, 7 cinnamic acid derivatives, 8 triterpenoids, and 14 others were identified or tentatively characterized by high-resolution molecular mass, fragment ions, and collision cross-section values. Furthermore, high-definition MSE was used to identify six pairs of co-eluting isomers that could not be detected from conventional data-independent acquisition and fast data-directed analysis. This research strategy has a certain potential for the analysis of other compound formulae and lays the foundation for the study of traditional Chinese medicine efficacy.

Integration of metabolomics and transcriptomics to reveal anti-chronic myocardial ischemia mechanism of Gualou Xiebai decoction

Ethnopharmacological relevanceGualou Xiebai decoction (GLXB), a well-known classic traditional Chinese medicine formula, is a recorded and proven therapy for the management of cardiac diseases. However, its pharmacological characteristics and mechanism of action are unclear. Materials and methodsThe effects of GLXB and its mechanism of action in an isoprenaline-induced rat model of chronic myocardial ischemia (CMI) were investigated by incorporating metabonomics and transcriptomics. Meanwhile, the echocardiographic evaluation, histopathological analysis, serum biochemistry assay, TUNEL assay and western blot analysis were detected to revealed the protective effects of GLXB on CMI. ResultsThe results of echocardiographic evaluation, histopathological analysis and serum biochemistry assay revealed that GLXB had a significantly cardioprotective performance by reversing echocardiographic abnormalities, restoring pathological disorders and converting the serum biochemistry perturbations. Further, the omics analysis indicated that many genes and metabolites were regulated after modeling and GLXB administration, and maintained the marked “high-low” or “low-high” trends. Meanwhile, the results from integrated bioinformatics analysis suggested that the interaction network mainly consisted of amino acid and organic acid metabolism. The results of TUNEL assay and western blot analysis complemented the findings of integrated analysis of metabolomics and transcriptomics. ConclusionThese findings suggested that GLXB has a curative effect in isoproterenol-induced CMI in rats. Integrated analysis based on transcriptomics and metabolomics studies revealed that the mechanism of GLXB in alleviating CMI was principally by the regulation of energy homeostasis and apoptosis, which was through a multi-component and multi-target treatment modality.

Herb-drug interaction between Shaoyao-Gancao-Fuzi decoction and tofacitinib via CYP450 enzymes

Ethnopharmacology relevanceShaoyao-Gancao-Fuzi decoction (SGFD), a well-known traditional Chinese medicine formula, was originally described in “Treatise on Febrile Diseases” and has been extensively used to dispel wind, eliminate dampness and treat paralysis. It is widely used for the treatment of rheumatoid arthritis in clinic. However, the effect of SGFD on the activity of cytochrome P450 enzymes (CYP450s) and the herb-drug interactions are rarely studied. ObjectiveThe aim of this study was to investigate the effect of SGFD on the activity of CYP450s and evaluate the potential herb-drug interactions between SGFD and tofacitinib, commonly used disease-modifying antirheumatic drug in rheumatoid arthritis. Materials and methodsThe cocktail approach was employed to assess the effect of SGFD on the activity of CYP1A2, 3A4, 2A6, 2E1, and 2C9. The pharmacokinetic profile of oral administration of tofacitinib in rats after two weeks of treatment with SGFD was investigated. RT-qPCR and molecular docking were performed to unveil the underlying mechanism of the herb-drug interaction. ResultsSGFD had no effect on the activities of CYP2E1 and 2C9, had a weak effect on CYP2A6, and had activatory effect on CYP1A2. However, it had a dramatically inhibitory effect on the activity of CYP3A4. Simultaneously, the values of Cmax and AUC0-∞ of tofacitinib were obviously increased after treatment with SGFD for 14 days. The mechanism study manifested that SGFD significantly reduced the gene transcription of CYP3A. Molecular docking work confirmed that the inhibitory activity of glycyrrhetinic acid, glycyrrhizic acid and liquiritin, the main ingredients of SGFD, occurred by occupying the active sites of CYP3A4 and by making favorable interactions with its key residues. ConclusionsThe system exposure of tofacitinib was increased by SGFD. SGFD could affect the activity and gene expression of the key metabolic enzyme CYP3A. These findings give a clear understanding to predict herb-drug interaction of SGFD for safe clinical use in future.

Establishing a chromatographic fingerprint using tandem UV/charged aerosol detection and similarity analysis for Shengmai capsule: A novel method for natural product quality control.

Shengmai San, a well-known traditional Chinese medicine formula, is used to treat coronary heart diseases and myocardial infarction. The complex composition and complicated mechanism of the Shengmai preparations bring a significant challenge in the development of a suitable quality control method. This work aims to establish a chromatographic fingerprinting method and propose a weighting algorithm for application in fingerprint similarity analysis to ensure consistent quality of the Shengmai capsule. A chromatographic fingerprint method was established using tandem UV/charged aerosol detection (CAD) for Shengmai capsule quality control. After method verification, the developed method was applied to analyze 15 batches of the samples. Then a weighting algorithm of the fingerprint peak was proposed and used for the fingerprint similarity analysis. An HPLC-UV/CAD fingerprint method was successfully developed for the Shengmai capsules. Chromatographic conditions of the HPLC-UV/CAD method were optimized with a definitive screening design, and the optimized ranges of operating parameters were obtained with a Monte Carlo simulation method. The combined use of the proposed weighting algorithm and similarity analysis on fingerprint data improves the sensitivity of distinguishing batch-to-batch quality differences. The developed HPLC-UV/CAD fingerprint method is robust, reliable, and efficient. The proposed weighting algorithm combined with similarity analysis is promising and meaningful for the quality consistency assessment of HPLC-UV/CAD fingerprints.

Anticancer Action of Xiaoxianxiong Tang in Non-Small Cell Lung Cancer by Pharmacological Analysis and Experimental Validation

Xiaoxianxiong Tang (XXXT) is a well-known traditional Chinese medicine formula. Evidence is emerging supporting the benefits of XXXT in ameliorating therapy for non-small cell lung cancer (NSCLC). The purpose of this study aimed to explore the effects and mechanisms of XXXT through network pharmacological analysis and biological validation. TCMSP database was used to identify potentially active compounds in XXXT with absorption, distribution, metabolism, excretion screening, and their potential targets. The disease targets related to NSCLC were predicted by searching for Therapeutic Target database, GeneCards database, DrugBank database, and DisGeNET database. Of the 4385 NSCLC-related targets, 156 targets were also the targets of compounds present in XXXT. Subsequently, GO function and KEGG pathway enrichment and PPI network analyses revealed that, of the 95 targets and 20 pathways influenced by 20 ingredients in XXXT, 20 targets were associated with patient survival, and XXXT could exert an inhibitory action on the PI3K-AKT signaling pathway. Moreover, XXXT restrained the proliferation of A549 and H460 cells in a concentration-dependent manner and suppressed the mRNA and protein levels of key targets CCNA2, FOSL2, and BIRC5 closely linked to the PI3K-AKT pathway. Hence, XXXT has the potential to improve therapy for NSCLC by targeting the PI3K-AKT signaling pathway.

Evaluation of the effect of Shengxian Decoction on doxorubicin-induced chronic heart failure model rats and a multicomponent comparative pharmacokinetic study after oral administration in normal and model rats

Shengxian Decotion (SXT), a well-known Traditional Chinese Medicine (TCM) formula composed of Astragali Radix, Bupleuri Radix, Cimicifugae Rhizoma, Anemarrhenae Rhizoma and Platycodonis Radix, is clinically considered as an effective formula against cardiovascular diseases. However, the exact effective substance of SXT in treating chronic heart failure (CHF) still remains unclear. In the current study, we investigated the benefit of SXT in doxorubicin (DOX)-induced CHF rats and established a UHPLC-MS/MS method to simultaneously determine 18 key compounds in a subsequent comparative pharmacokinetic study in normal and CHF rats. Histopathological studies, transmission electron microscopy, and echocardiography were applied to assess the therapeutic effect of SXT on DOX-induced CHF rats, which indicated that SXT significantly ameliorated DOX-induced CHF, similar to enalapril. In addition, we successfully established a UHPLC-MS/MS method to determine the pharmacokinetics of the components in rat plasma, which was validated with good linearity, inter-day and intra-day precisions and accuracies, matrix effects, extraction recovery, and stability values. Our results showed that only astragaloside IV showed increased plasma exposure in the CHF rats, while saikosaponin A, quercetin, timosaponin B-II, ferulic acid, isoferulic acid and formononetin decreased compared to their pharmacokinetic characteristics in the normal and CHF rats. This study demonstrates that SXT enjoys obvious therapeutic effect on DOX-induced CHF rats, and the altered metabolism of some compounds in SXT is affected by the pathological state of CHF rats. Our findings provide a better understanding of the in vivo exposure to complex compounds of SXT, supporting effective substance screening and further investigation of the therapeutic mechanism.

Qingda Granule Attenuates Angiotensin II-Induced Blood Pressure and Inhibits Ca2+/ERK Signaling Pathway.

Objective: As a well-known traditional Chinese medicine formula prescribed by academician Ke-ji Chen, Qingda granule (QDG) lowered the blood pressure of spontaneously hypertensive rats and attenuated hypertensive cardiac remodeling and inflammation. However, its functional role and underlying mechanisms on hypertensive vascular function remain largely unclear. This study aims to assess the effects of QDG treatment on Angiotensin II- (AngII-) induced hypertension and vascular function and explore its underlying mechanisms both in vitro and in vivo.Methods: In an in vivo study, 25 male C57BL/6 mice were randomly divided into five groups, including Control, AngII, AngII + QDG-L, AngII + QDG-M, and AngII + QDG-H groups (n = 5 for each group). Mice in AngII and AngII + QDG-L/-M/-H groups were infused with AngII (500 ng/kg/min), while in the Control group, they were infused with saline. Mice in AngII + QDG were intragastrically given different concentrations of QDG (0.5725, 1.145, or 2.29 g/kg/day), while in Control and AngII groups, they were intragastrically given equal volumes of double distilled water for 2 weeks. Blood pressure was determined at 0, 1, and 2 weeks of treatment. Ultrasound was used to detect the pulse wave velocity (PWV) and HE staining to detect the pathological change of the abdominal aorta. RNA sequencing (RNA-seq) was performed to identify the differentially expressed transcripts (DETs) and related signaling pathways. IHC was used to detect the expression of p-ERK in the abdominal aorta. Primary isolated rat vascular smooth muscle cells (VSMCs) were used to assess the cellular Ca2+ release and activation of the ERK pathway by confocal microscope and western blotting analysis, respectively.Results: QDG treatment significantly alleviated the elevated blood pressure, the PWV, and the thickness of the abdominal aorta in AngII-induced hypertensive mice. RNA-seq and KEGG analyses identified 1,505 DETs and multiple enriched pathways (including vascular contraction and calcium signaling pathway) after QDG treatment. Furthermore, confocal microscope showed that QDG treatment partially attenuated the increase of Ca2+ release with the stimulation of AngII in cultured VSMCs. In addition, IHC and western blotting indicated that QDG treatment also partially alleviated the increase of phospho-ERK levels in abdominal aorta tissues of mice and cultured VSMCs after the infusion or stimulation of AngII.Conclusion: QDG treatment attenuated the elevation of blood pressure, abdominal aorta dysfunction, pathological changes, Ca2+ release, and activation of the ERK signaling pathway.

Bawei Chenxiang Wan Ameliorates Cardiac Hypertrophy by Activating AMPK/PPAR-α Signaling Pathway Improving Energy Metabolism.

Bawei Chenxiang Wan (BCW), a well-known traditional Chinese Tibetan medicine formula, is effective for the treatment of acute and chronic cardiovascular diseases. In the present study, we investigated the effect of BCW in cardiac hypertrophy and underlying mechanisms. The dose of 0.2, 0.4, and 0.8 g/kg BCW treated cardiac hypertrophy in SD rat model induced by isoprenaline (ISO). Our results showed that BCW (0.4 g/kg) could repress cardiac hypertrophy, indicated by macro morphology, heart weight to body weight ratio (HW/BW), left ventricle heart weight to body weight ratio (LVW/BW), hypertrophy markers, heart function, pathological structure, cross-sectional area (CSA) of myocardial cells, and the myocardial enzymes. Furthermore, we declared the mechanism of BCW anti-hypertrophy effect was associated with activating adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK)/peroxisome proliferator–activated receptor-α (PPAR-α) signals, which regulate carnitine palmitoyltransferase1β (CPT-1β) and glucose transport-4 (GLUT-4) to ameliorate glycolipid metabolism. Moreover, BCW also elevated mitochondrial DNA-encoded genes of NADH dehydrogenase subunit 1(ND1), cytochrome b (Cytb), and mitochondrially encoded cytochrome coxidase I (mt-co1) expression, which was associated with mitochondria function and oxidative phosphorylation. Subsequently, knocking down AMPK by siRNA significantly can reverse the anti-hypertrophy effect of BCW indicated by hypertrophy markers and cell surface of cardiomyocytes. In conclusion, BCW prevents ISO-induced cardiomyocyte hypertrophy by activating AMPK/PPAR-α to alleviate the disturbance in energy metabolism. Therefore, BCW can be used as an alternative drug for the treatment of cardiac hypertrophy.

Exploring the Active Components of Simotang Oral Liquid and Their Potential Mechanism of Action on Gastrointestinal Disorders by Integrating Ultrahigh-Pressure Liquid Chromatography Coupled with Linear Ion Trap-Orbitrap Analysis and Network Pharmacology.

Simotang oral liquid (SMT), a well-known traditional Chinese medicine formula composed of four medicinal and edible plants, has been extensively used for treating gastrointestinal disorders (GIDs) since ancient times. However, the major active constituents and the underlying molecular mechanism of SMT on GIDs are still partially understood. Herein, the preliminary chemical profile of SMT was first identified by ultrahigh-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry (UHPLC-LTQ-Orbitrap). In total, 70 components were identified. Then, a network pharmacology approach integrating target prediction, pathway enrichment analysis, and network construction was adopted to explore the therapeutic mechanism of SMT. As a result, 170 main targets were screened out and considered as effective players in ameliorating GIDs. More importantly, the major hubs were found to be highly enriched in a calcium signaling pathway. Furthermore, 26 core SMT-related genes were identified, which may play key roles in ameliorating gastrointestinal motility. In conclusion, this work would provide valuable information for further development and clinical application of SMT.

Qingda granule attenuates angiotensin II-induced cardiac hypertrophy and apoptosis and modulates the PI3K/AKT pathway

Qingda granule (QDG), simplified from Qingxuan Jiangya Decoction, is a well-known traditional Chinese medicine formula that has been used for decades to treat hypertension. However, the cardioprotective effects of QDG on Ang II-induced hypertension remain unknown. This study aimed to investigate the effects of QDG on hypertension-induced cardiac hypertrophy and apoptosis, as well as explore its underlying mechanisms. Mice were infused with Ang II (500 ng/kg/min) or saline solution as control, then administered oral QDG (1.145 g/kg/day) or saline for two weeks. QDG treatment attenuated the elevation in blood pressure caused by Ang II, as well as the decreased left ventricle ejection fractions and fractional shortening. Moreover, QDG treatment significantly alleviated the Ang II-induced elevation of the ratio of heart weight to tibia length, as well as cardiac injury, hypertrophy, and apoptosis. In cultured H9C2 cells stimulated with Ang II, QDG partially reversed the increase in cell surface area and number of apoptotic cells, up-regulation of hypertrophy markers ANP and BNP, and activation of caspases-9 and -3. QDG also partially reversed Ang II-induced accumulation of reactive oxygen species (ROS), depolarization of the mitochondrial membrane, release of cytochrome C, up-regulation of Bax, and decrease in levels of p-PI3K, p-AKT, and Bcl-2. These results suggest that QDG can significantly attenuate Ang II-induced hypertension, cardiac hypertrophy and apoptosis, and it may exert these effects in part by suppressing ROS production and activating the PI3K/AKT signaling pathway.

Effect of shengmai san on insulin resistance, tumor necrosis factor-alpha and oxidative stress in rats fed on a high-fat diet

Shengmai San (SMS) is a well-known traditional Chinese medicine formula prescribed for patients with coronary heart disease (CHD). Insulin resistance is the most likely explanation for the development and progression of this disease. To investigate the effect of SMS on high-fat diet-induced insulin resistance, inflammatory cytokine production and oxidative stress, male Wistar rats were fed with low-fat control diet, high-fat diet and high-fat diet supplemented with 4% SMS for eight weeks. An oral glucose tolerance test was conducted in the seventh week. Rats fed the SMS diet had significantly lower plasma fructosamine concentration (p < 0.05) and tended to have lower (p < 0.1) AUC values of plasma glucose and insulin concentrations after glucose challenge, compared to those fed the high-fat diet. Moreover, SMS reduced TNF-α and lipid peroxidation levels in the liver and heart. However, SMS had no effect on fasting plasma concentrations of glucose, insulin, triglyceride, non-esterified fatty acids, total cholesterol, HDL-cholesterol and insulin resistance index (homeostasis model assessment; HOMA) in rats. Our results show that SMS may have little or no significant effect on reducing insulin resistance, but display anti-oxidative and anti-inflammatory properties.