Advanced Glycation End Products Signaling Research Articles

The Identification of Potential Drugs for Dengue Hemorrhagic Fever: Network-Based Drug Reprofiling Study

Background Dengue fever can progress to dengue hemorrhagic fever (DHF), a more serious and occasionally fatal form of the disease. Indicators of serious disease arise about the time the fever begins to reduce (typically 3 to 7 days following symptom onset). There are currently no effective antivirals available. Drug repurposing is an emerging drug discovery process for rapidly developing effective DHF therapies. Through network pharmacology modeling, several US Food and Drug Administration (FDA)-approved medications have already been researched for various viral outbreaks. Objective We aimed to identify potentially repurposable drugs for DHF among existing FDA-approved drugs for viral attacks, symptoms of viral fevers, and DHF. Methods Using target identification databases (GeneCards and DrugBank), we identified human–DHF virus interacting genes and drug targets against these genes. We determined hub genes and potential drugs with a network-based analysis. We performed functional enrichment and network analyses to identify pathways, protein-protein interactions, tissues where the gene expression was high, and disease-gene associations. Results Analyzing virus-host interactions and therapeutic targets in the human genome network revealed 45 repurposable medicines. Hub network analysis of host-virus-drug associations suggested that aspirin, captopril, and rilonacept might efficiently treat DHF. Gene enrichment analysis supported these findings. According to a Mayo Clinic report, using aspirin in the treatment of dengue fever may increase the risk of bleeding complications, but several studies from around the world suggest that thrombosis is associated with DHF. The human interactome contains the genes prostaglandin-endoperoxide synthase 2 (PTGS2), angiotensin converting enzyme (ACE), and coagulation factor II, thrombin (F2), which have been documented to have a role in the pathogenesis of disease progression in DHF, and our analysis of most of the drugs targeting these genes showed that the hub gene module (human-virus-drug) was highly enriched in tissues associated with the immune system (P=7.29 × 10–24) and human umbilical vein endothelial cells (P=1.83 × 10–20); this group of tissues acts as an anticoagulant barrier between the vessel walls and blood. Kegg analysis showed an association with genes linked to cancer (P=1.13 × 10–14) and the advanced glycation end products–receptor for advanced glycation end products signaling pathway in diabetic complications (P=3.52 × 10–14), which indicates that DHF patients with diabetes and cancer are at risk of higher pathogenicity. Thus, gene-targeting medications may play a significant part in limiting or worsening the condition of DHF patients. Conclusions Aspirin is not usually prescribed for dengue fever because of bleeding complications, but it has been reported that using aspirin in lower doses is beneficial in the management of diseases with thrombosis. Drug repurposing is an emerging field in which clinical validation and dosage identification are required before the drug is prescribed. Further retrospective and collaborative international trials are essential for understanding the pathogenesis of this condition.

Epithelial to mesenchymal transition in mammary gland tissue fibrosis and insights into drug therapeutics.

The epithelial-mesenchymal transition (EMT) is a multi-step morphogenetic process in which epithelial cells lose their epithelial properties and gain mesenchymal characteristics. The process of EMT has been shown to mediate mammary gland fibrosis. Understanding how mesenchymal cells emerge from an epithelial default state will aid in unravelling the mechanisms that control fibrosis and, ultimately, in identifying therapeutic targets to alleviate fibrosis. The effects of EGF and high glucose (HG) on EMT in mammary epithelial cells, MCF10A and GMECs, as well as their pathogenic role, were studied. In-silico analysis was used to find interacting partners and protein-chemical/drug molecule interactions. On treatment with EGF and/or HG, qPCR analysis showed a significant increase in the gene expression of EMT markers and downstream signalling genes. The expression of these genes was reduced on treatment with EGF+HG combination in both cell lines. The protein expression of COL1A1 increased as compared to the control in cells treated with EGF or HG alone, but when the cells were treated with EGF and HG together, the protein expression of COL1A1 decreased. ROS levels and cell death increased in cells treated with EGF and HG alone, whereas cells treated with EGF and HG together showed a decrease in ROS production and apoptosis. In-silico analysis of protein-protein interactions suggest the possible role of MAPK1, actin alpha 2 (ACTA2), COL1A1, and NFκB1 in regulating TGFβ1, ubiquitin C (UBC), specificity protein 1 (SP1) and E1A binding protein P300 (EP300). Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment suggests advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signalling pathway, relaxin signalling pathway and extra cellular matrix (ECM) receptor interactions underlying fibrosis mechanism. This study demonstrates that EGF and HG induce EMT in mammary epithelial cells and may also have a role in fibrosis.

Study on the pharmacological mechanisms of sodium-glucose co-transporter 2 inhibitors in obesity-related atrial fibrillation based on network pharmacology and experimental verification.

Obesity is an independently risk factor of atrial fibrillation (AF). It is likely that the global burden of AF will escalate due to the current obesity epidemic. Weight loss can effectively reduce the risk of AF, while sodium-glucose co-transporter 2 inhibitors (SGLT2i) can reduce body weight, so SGLT2i are potentially effective for obesity-related AF. SGLT2i are a novel type of oral medication. This current study used network pharmacology to examine the potential mechanisms of SGLT2i in the treatment of obesity-related AF, and the therapeutic effects were assessed in vivo. Potential gene targets for SGLT2i in treating obesity-related AF were identified from public database. Cytoscape V3.7.1 was used to construct the "Drug-Target" and "Drug-Target-Disease" networks. The STRING database was applied to investigate the protein-protein interactions (PPIs). Additionally, the Bioconductor tools were used to analyze the Gene Ontology (GO) biological functions and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The efficacy of SGLT2i in treating obesity-related AF was investigated in vivo using a diet-induced obese C57BL/6J male mouse model. A number of indicators were assessed, including invasive electrophysiology, testing of blood samples, and expression detection of pathway targets. These experiments were used to verify the targets mined by network pharmacology. The results indicated that SGLT2i had 80 potential target genes during the treatment of obesity-related AF, and 10 hub genes were obtained by further screening. It was predicted that the treatment of obesity-related AF by SGLT2i involved the advanced glycation end product (AGE)-receptor for advanced glycation end product (RAGE) signaling pathway and other signaling pathways. In in vivo experiments, administration of SGLT2i (the SGLT2i + DIO group) resulted in a lower AF induction rate (P<0.05), decreased serum AGEs/soluble RAGE (sRAGE) ratio (P<0.01), and decreased expression of NADPH oxidase 2 (NOX2) (P<0.05) compared to untreated DIO mice (the DIO group). In this study, pharmacological network analysis and in vivo experiments demonstrated that SGLT2i acts on obesity-related AF by inhibiting the AGE-RAGE signaling pathway. These results offer fresh perspectives on the pharmacological actions of SGLT2i in treating obesity-related AF.

Stroke-induced hexokinase 2 in circulating monocytes exacerbates vascular inflammation and atheroprogression

BackgroundStroke accelerates inflammatory monocyte recruitment to the endothelium and consequent atheroprogression via high-mobility group box 1-receptor for advanced glycation end products signaling. Notably, Hmgb1 interacts with multiple toll-like receptors (TLRs) and promotes TLR4-mediated proinflammatory myeloid cell activation. Therefore, TLR-associated mechanism(s) within monocytes may play a role in Hmgb1-driven poststroke atheroprogression. ObjectivesWe aimed to elucidate the TLR-associated mechanism(s) within monocytes that contribute to stroke-induced exacerbation of atherosclerotic disease. MethodsA weighted gene coexpression network analysis on the whole blood transcriptomes of stroke model mice identified hexokinase 2 (HK2) as a key gene associated with TLR signaling in ischemic stroke. We conducted a cross-sectional analysis of monocyte HK2 levels in patients with ischemic stroke patients. We performed in vitro and in vivo studies using high-cholesterol diet–fed myeloid-specific Hk2-null ApoE−/− (ApoE−/−;Hk2ΔMφ) mice and ApoE−/−;Hk2fl/fl controls. ResultsWe found markedly higher monocyte HK2 levels in patients with ischemic stroke patients during the acute and subacute phases poststroke. Similarly, stroke model mice displayed a profound increase in monocyte Hk2 levels. Using aortas and aortic valve samples collected from high-cholesterol diet–fed ApoE−/−;Hk2ΔMφ mice and ApoE−/−;Hk2fl/fl controls, we found that stroke-induced monocyte Hk2 upregulation enhanced poststroke atheroprogression and inflammatory monocyte recruitment to the endothelium. Stroke-induced monocyte Hk2 upregulation induced inflammatory monocyte activation, systemic inflammation, and atheroprogression via Il-1β. Mechanistically, we demonstrated that stroke-induced monocyte Hk2 upregulation was dependent upon Hmgb1-driven p38-dependent hypoxia-inducible factor-1α stabilization. ConclusionStroke-induced monocyte Hk2 upregulation is a key mechanism underlying poststroke vascular inflammation and atheroprogression.

Wumei Wan attenuates angiogenesis and inflammation by modulating RAGE signaling pathway in IBD: Network pharmacology analysis and experimental evidence

BackgroundWumei Wan (WMW) has been used to address digestive disorder for centuries in traditional Chinese medicine. Previous studies have demonstrated its anti-colitis efficacy, but the underlying mechanism of its action remains to be further clarified. PurposeTo investigate the underlying mechanisms of WMW in the treatment of chronic ulcerative colitis (UC) through network pharmacology and experimental validation. MethodsTraditional Chinese Medicine Systems Pharmacology (TCMSP) platform were used to identify the ingredients and potential targets of WMW. The microarray gene data GSE75214 datasets from GEO database was used to define UC-associated targets. Cytoscape3.7.2 was employed to construct the protein-protein interaction (PPI) network and compounds-disease targets network. GO enrichment analysis and KEGG pathway analysis were performed by R software for functional annotation. UPLC-TOF-MS/MS method was used to quantitatively analyze the active ingredients of WMW. For experimental validation, three cycles of 2% dextran sulfate sodium salt (DSS) were used to construct chronic colitis model. The hub targets and signal pathway were detected by qPCR, ELISA, western blotting , immunohistochemical and immunofluorescence. ResultsThrough network analysis, 104 active ingredients were obtained from WMW, and 47 of these ingredients had potential targets for UC. A total of 41 potential targets of WMW and 13 hub targets were identified. KEGG analysis showed that WMW involved in advanced glycation end products-receptor of advanced glycation end products (AGE-RAGE) signaling pathway. Taxifolin, rutaecarpine, kaempferol, quercetin, and luteolin of WMW were the more highly predictive components related to the AGE-RAGE signaling pathway. In vivo validation, WMW improved DSS-induced colitis, reduced the expression of inflammatory cytokines and chemokines. Notably, it significantly decreased the mRNA expression of Spp1, Serpine1, Mmp2, Mmp9, Ptgs2, Nos2, Kdr and Icam1, which were associated with angiogenesis. In addition, we confirmed WMW inhibited RAGE expression and diminished DSS-induced epithelial barrier alterations ConclusionOur results initially demonstrated the effective components and the strong anti-angiogenic activity of WMW in experimental chronic colitis. Sufficient evidence of the satisfactory anti-colitis action of WMW was verified in this study, suggesting its potential as a quite prospective agent for the therapy of UC.

Therapeutic targets and signal transduction mechanisms of medicinal plant formula Gancao Xiexin decoction against ulcerative colitis: A network pharmacological study

<b>Background:</b> Ulcerative colitis (UC) is a chronic disease that often presents with abdominal pain, diarrhea, hematochezia, and significant morbidity. Gancao Xiexin decoction (GXD), a traditional Chinese medicine, has been applied for the clinical treatment of UC, while its action mechanisms are unclear. <b>Methods:</b> The active ingredients and their targets of GXD, and UC-related targets, were derived from public databases. Protein-protein interaction, Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the important active compounds, key targets, and signaling pathways. Then, molecular docking and animal experiments were performed to verify the findings. A total of 213 active compounds and 89 common targets of GXD for UC were obtained. <b>Results:</b> The hub gene network showed <i>ALB</i>, <i>AKT1</i>, <i>IL6</i>, <i>TNF</i>, <i>VEGFA</i>, <i>TP53</i>, <i>CXCL8</i>, <i>MAPK1</i>, <i>PTGS2</i>, and <i>IL1β</i> may be potential targets of GXD against UC. GO and KEGG pathway enrichment analyses suggested that the action of GXD against UC was mainly related to response to oxygen levels, lipopolysaccharide, and molecule of bacterial origin, etc., and achieved by advanced glycation endproducts/receptors for advanced glycation endproducts signaling pathway in diabetic complications, hypoxia-inducible factor (HIF)-1 signaling pathway, interleukin-17/HIF-1 signaling pathway, TNF signaling pathway, etc. Molecular docking results showed that the GXD had good potency of action with the hub target. <i>In vivo</i> experiments showed that GXD significantly alleviated the symptoms of UC and down-regulated the expression of inflammatory factors, nuclear factor-κB and signal transducer and activator of transcription 3. <b>Conclusions:</b> The anti-UC action of GXD is mainly attributed to its anti-oxidative stress, anti-inflammatory, and immunomodulatory functions.

Advanced glycation end products modulate electrophysiological remodeling of right ventricular outflow tract cardiomyocytes: A novel target for diabetes-related ventricular arrhythmogenesis.

Diabetes mellitus is associated with cardiovascular disease and cardiac arrhythmia. Accumulation of advanced glycation end products closely correlates with cardiovascular complications through mitochondrial dysfunction or oxidative stress and evoke proliferative, inflammatory, and fibrotic reactions, which might impair cardiac electrophysiological characteristics and increase the incidence of cardiac arrhythmia. This study examined the mechanisms how advanced glycation end products may contribute to arrhythmogenesis of right ventricular outflow tract-a unique arrhythmogenic substrate. A whole-cell patch clamp, conventional electrophysiological study, fluorescence imaging, Western blot, and confocal microscope were used to study the electrical activity, and Ca2+ homeostasis or signaling in isolated right ventricular outflow tract myocytes with and without advanced glycation end products (100 μg/ml). The advanced glycation end products treated right ventricular outflow tract myocytes had a similar action potential duration as the controls, but exhibited a lower L-type Ca2+ current, higher late sodium current and transient outward current. Moreover, the advanced glycation end products treated right ventricular outflow tract myocytes had more intracellular Na+ , reverse mode Na+ -Ca2+ exchanger currents, intracellular and mitochondrial reactive oxygen species, and less intracellular Ca2+ transient and sarcoplasmic reticulum Ca2+ content with upregulated calcium homeostasis proteins and advanced glycation end products related signaling pathway proteins. In conclusions, advanced glycation end products modulate right ventricular outflow tract electrophysiological characteristics with larger late sodium current, intracellular Na+ , reverse mode Na+ -Ca2+ exchanger currents, and disturbed Ca2+ homeostasis through increased oxidative stress mediated by the activation of the advanced glycation end products signaling pathway.

Single-cell transcriptomic, transcriptomic, and metabolomic characterization of human atherosclerosis

Atherosclerosis is the main cause of many cardiovascular and cerebrovascular diseases (CVDs), and gaining a deeper understanding of the intercellular connections and key central genes which mediate formation of atherosclerotic plaques is required. We performed a comprehensive bioinformatics analysis of differential genetic screening, Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway annotation, protein-protein interactions (PPIs), pseudo-timing, intercellular communication, transcription factors on carotid single-cell sequencing data, and aortic bulk transcriptome and metabolomic data. Ten cell types were identified in the data: T cells, monocytes, smooth muscle cells, endothelial cells, B cells, fibroblasts, plasma cells, mast cells, dendritic cells, and natural killer cells. Endothelial, fibroblast, macrophage, and smooth muscle cell subtype differentiation trajectories, interaction networks, and important transcription factors were characterized in detail. Finally, using this information combined with transcriptome and metabolome analyses, we found the key genes and signaling pathways of atherosclerosis, especially the advanced glycation end products and receptor for advanced glycation end products signaling pathway (AGE-RAGE signaling pathway) in diabetic complications, linked the differential metabolites with fibroblasts and atherosclerosis and contributed to it in patients with diabetes. Collectively, this study provides key genes, signaling pathways, cellular communication, and transcription factors among endothelial cells, fibroblasts, macrophages, and smooth muscle cells for the study of atherosclerotic plaques, and provides a basis for the diagnosis and treatment of atherosclerosis-like sclerosis.

Glycation-Associated Diabetic Nephropathy and the Role of Long Noncoding RNAs.

The glycation of various biomolecules is the root cause of many pathological conditions associated with diabetic nephropathy and end-stage kidney disease. Glycation imbalances metabolism and increases renal cell injury. Numerous therapeutic measures have narrowed down the adverse effects of endogenous glycation, but efficient and potent measures are miles away. Recent advances in the identification and characterization of noncoding RNAs, especially the long noncoding RNAs (lncRNAs), have opened a mammon of new biology to explore the mitigations for glycation-associated diabetic nephropathy. Furthermore, tissue-specific distribution and condition-specific expression make lncRNA a promising key for second-generation therapeutic interventions. Though the techniques to identify and exemplify noncoding RNAs are rapidly evolving, the lncRNA study encounters multiple methodological constraints. This review will discuss lncRNAs and their possible involvement in glycation and advanced glycation end products (AGEs) signaling pathways. We further highlight the possible approaches for lncRNA-based therapeutics and their working mechanism for perturbing glycation and conclude our review with lncRNAs biology-related future opportunities.

Network Pharmacology-Based Study on the Efficacy and Mechanism of Lonicera japonica Thunberg

Network pharmacology is an emerging method for investigating the potential effects and mechanisms of natural products through system-level analyses of gene sets in herbs. Lonicera japonica Thunberg (LJ) is known to have anti-inflammatory, anti-bacterial, anti-oxidant, anti-tumor and neuroprotective effects. In the present study, network pharmacological analysis was performed to assess the potential efficacy and mechanisms of LJ. First of all, constituents of LJ were gathered from public databases: the Oriental Advanced Searching Integrated System (OASIS) database, PubChem and the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database. Then, a network was constructed using Cytoscape3.8.2, which visualizes biomedical interactions, and a functional enrichment analysis was conducted to uncover the pathways most relevant to LJ through Enrichr based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway 2021. Further, we performed a study of the literature to determine whether the results of our study were consistent with those of previous studies. The results showed that ‘Advanced glycation end products-Receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications’ was the pathway most relevant to LJ, especially through ‘Mitogen-activated protein kinase (MAPK) signaling pathway’, ‘Phosphatidylinositol 3 kinase-Protein kinase B (PI3K-AKT) signaling pathway’ and ‘Janus kinase-Signal transducers and activators of transcription (JAK-STAT) signaling pathway’. Based on the literature study, LJ showed relevance to MAPK, PI3K-AKT and JAK-STAT and was associated with therapeutic effects on diabetes and diabetic complications. This study shows that network pharmacology can be a suitable approach for analyzing LJ and suggests the potential efficacy and mechanisms of LJ.

Study on the mechanism of Ginseng-Gegen for mesenteric lymphadenitis based on network pharmacology.

BackgroundThis study aimed to determine the main active ingredients of the Ginseng-Gegen (Panax Ginseng-Radix Puerariae) drug pair, to predict relevant action targets, and to establish a network of “drug-active ingredients-targets”, to ultimately explore the mechanism of Ginseng-Gegen in the treatment of mesenteric lymphadenitis.MethodsThe Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform was used to screen the chemical constituents of Ginseng-Gegen, and the active ingredient targets were retrieved by UniProt database. The databases of GeneCards and the Online Mendelian Inheritance in Man (OMIM) were applied to search for mesenteric lymphadenitis-related targets. Cytoscape software was used to construct the network of active ingredient-action targets. The biological functions of the targets were analyzed in the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database.ResultsA total of 26 potential active ingredients of the Ginseng-Gegen drug pair were screened, with 128 drug-related targets and 255 mesenteric lymphadenitis-related targets. After matching, 23 potential targets were obtained for treating mesenteric lymphadenitis. Among them, MOL012297 (puerarin), MOL005344 (ginsenoside Rh2), and MOL000358 (beta-sitosterol) were linked to 3 or more key target genes. They were supposed to be important ingredients of Ginseng-Gegen in the treatment of mesenteric lymphadenitis.ConclusionsGinseng-Gegen is related to oxidative stress and inflammation, and it is a part of the nuclear factor κB (NF-κB) signaling pathway, tumor necrosis factor (TNF) signaling pathway, and the advanced glycation end products/receptor for advanced glycation end products (AGE-RAGE) signaling pathway. These biological processes and signaling pathways may be potential mechanisms of Ginseng-Gegen for treating mesenteric lymphadenitis.

Abstract A3: Possible Differential Effects Of Angiotensin Receptor-neprilysin Inhibitor And Angiotensin Receptor Blocker Against Hypertension And Cardiorenal Injury In A Mouse Model Of Cardiorenal Syndrome

Objective: Angiotensin receptor-neprilysin inhibitors (ARNI) are potential therapeutic candidates for the treatment of cardiorenal syndrome (CRS). Although evidence is accumulating on the therapeutic effects of ARNI for heart failure and hypertension, the effects of ARNI on kidney disease or CRS are controversial. Here, we assessed the hypothesis that ARNI has poor renal protective effect beyond its antihypertensive effect, in contrast to its promised cardioprotective effect, by using the ANS [Ang II (A) + nephrectomy (N) + saline (S)] mice, which is the CRS model. Method: C57BL/6 mice were divided into 5 groups (N = 5-6): Control (sham operation) group, ANS (ANS + vehicle treatment) group, Val M [ANS + moderate dose of valsartan treatment (30 mg/kg/day)] group, ARNI [ANS + sacubitril/valsartan treatment (60 mg/kg/day)] group, and Val H [ANS + high dose of valsartan treatment (60 mg/kg/day)] group. Sacrifice was performed after 4 weeks, and various analyses were conducted. Results: ARNI, Val M, and Val H groups attenuated the elevation of blood pressure (BP) in ANS mice (BP [mean ± standard error (SEM)] [mmHg]: ANS, 164 ± 3; ARNI, 133 ± 4; Val M, 143 ±1; Val H, 129 ± 2). Regardless of the strength of the antihypertensive effect, the ARNI group was most effective in preventing the decrease in cardiac contraction and cardiac fibrosis in ANS mice. However, the ARNI group was insufficient to suppress urinary albumin excretion (UAE) (UAE [mean ± SEM] [μg]; ANS, 5351 ± 2391; ARNI, 1169 ± 700) and renal fibrosis in ANS mice. Both valsartan groups suppressed UAE in ANS mice (UAE [mean ± SEM] [μg]; Val M, 243 ± 141; Val H, 305 ± 179), as well as renal fibrosis and glomerular swelling. In addition, the results of kidney RNA sequencing analysis showed that the receptor for advanced glycation end-products (RAGE) signaling pathway was activated in the kidney of ARNI group compared with the Val M and Val H groups. Conclusions: The ARNI group showed better cardioprotective effects than valsartan treatment groups, but not sufficient kidney protection. These results suggested that the relationship between the antihypertensive effects and the organ protective effects of cardiorenal injury in CRS pathology may be different between angiotensin II receptor blocker (ARB) and ARNI.

Network pharmacology combined with pharmacodynamics revealed the anti-inflammatory mechanism of Tanreqing capsule against acute-exacerbation chronic obstructive pulmonary disease

Acute-exacerbation chronic obstructive pulmonary disease (AECOPD) is mainly associated with acute respiratory tract infection. In recent years, a growing number of studies have found that Tanreqing capsule (TRQ) has a favorable anti-inflammatory effect. In this study, we used network pharmacology and pharmacodynamics to explore the molecular mechanism and effects of TRQ in AECOPD treatment. To further understand the molecular mechanism of TRQ in AECOPD treatment, we used the network pharmacology to predict components of TRQ, TRQ-related targets, AECOPD-related targets, and pathways. In addition, we used the cigarette-smoke/lipopolysaccharide -induced AECOPD experimental model in Sprague–Dawley rats (72 rats randomly divided into six groups [n = 12 each]: control, model, high-TRQ [TRQ-H], medium-TRQ [TRQ-M], low-TRQ, and dexamethasone [Dex]) to evaluate the therapeutic effects of TRQ and to verify the network pharmacology. We found that 59 overlapping targets based on component-and AECOPD-related targets were frequently involved in the advanced glycation end product–receptor for advanced glycation end product signaling pathway in diabetic complications, the phosphatidylinositol-3-kinase–protein kinase B signaling pathway, and the hypoxia-inducible factor 1 signaling pathway, which might play important roles in the anti-inflammatory mechanism of TRQ in AECOPD treatment. Moreover, TRQ groups exerted protective effects against AECOPD by reducing the infiltration of inflammatory cells. Meanwhile, TRQ-M and TRQ-H groups significantly downregulated or upregulated the expression of tumor necrosis factor, interleukin (IL) 6, C-reactive protein, IL10, and serum amyloid A, as key targets in network pharmacology, in the serum and bronchoalveolar lavage fluid to achieve anti-inflammatory efficacy. Our study showed that TRQ had better anti-inflammatory efficacy against AECOPD, and initially elucidated its molecular mechanism. Moreover, our study also provides a new strategy to explore effective mechanism of TRQ against AECOPD; and further studies are needed to validate the biological processes and pathways of TRQ against AECOPD.

Reduning Attenuates LPS-Induced Human Unmilical Vein Endothelial Cells (HUVECs) Apoptosis Through PI3K-AKT Signaling Pathway.

The molecular mechanism of Reduning (RDN) in the treatment of sepsis was analyzed based on network pharmacology. The system pharmacology method was administered to search the active ingredients and targets of RDN, identify the sepsis-related genes, and determine the targets of RDN in the treatment of sepsis. Cytoscape was used to build a “drug component-target” network to screen key compounds. A protein-protein interaction (PPI) network was constructed using STRING, and core targets were revealed through topological analysis. 404 shared targets of RDN and sepsis were introduced into DAVID Bioinformatics Resources 6.8 for GO and KEGG enrichment analysis to predict their possible signaling pathways and explore their molecular mechanisms. GO enrichment analysis highlighted that they were largely related to protein phosphorylation, inflammatory reaction, and positive regulation of mitogen-activated protein kinase (MAPK) cascade. KEGG enrichment analysis outlined that they were enriched in PI3K-AKT signaling pathway, calcium signaling pathway, rhoptry-associated protein 1 (Rap1) signaling pathway, and advanced glycation end products and receptors for advanced glycation end products (AGE-RAGE) signaling pathway. Molecular biological validation results exposed that RDN could significantly improve the protein expression of p-AKT and p-PI3K, alleviate apoptosis-related proteins expression level and decrease apoptosis rate in LPS-induced HUVECs. In conclusion, it was illustrated that RDN could considerably constrain LPS-induced apoptosis by activating the PI3K-AKT signaling pathway, which advocated a basis for fundamental mechanism research and clinical application of RDN in the treatment of sepsis.

Effects of diabetes on osteocytes.

Better understanding of the mechanisms underlying skeletal dysfunction in the context of diabetes is needed to guide the development of therapeutic interventions to reduce the burden of diabetic fractures. Osteocytes, the 'master regulators' of bone remodeling, have emerged as key culprits in the pathogenesis of diabetes-related skeletal fragility. Both type 1 diabetes and type 2 diabetes cause chronic hyperglycemia that, over time, reduces bone quality and bone formation. In addition to acting as mechanosensors, osteocytes are important regulators of osteoblast and osteoclast activities; however, diabetes leads to osteocyte dysfunction. Indeed, diabetes causes the accumulation of advanced glycation end-products and senescent cells that can affect osteocyte viability and functions via increased receptor for advanced glycation endproducts (RAGE) signaling or the production of a pro-inflammatory senescence-associated secretory phenotype. These changes may increase osteocyte-derived sclerostin production and decrease the ability of osteocytes to sense mechanical stimuli thereby contributing to poor bone quality in humans with diabetes. Osteocyte dysfunction exists at the nexus of diabetic skeletal disease. Therefore, interventions targeting the RAGE signaling pathway, senescent cells, and those that inhibit sclerostin or mechanically stimulate osteocytes may alleviate the deleterious effects of diabetes on osteocytes and bone quality.

Mechanism underlying efficacy of Shugan Sanjie decoction on plasma cell mastitis, based on network pharmacology and experimental verification.

To investigate the mechanism underpinning the effeicay of Shugan Sanjie decoction (, SGSJD) on plasma cell mastitis (PCM) based on network pharmacology, and to verify it through . Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine were used to screen effective compounds and drug targets; Online Mendelian Inheritance in Man and GeneCards were used to search for PCM targets. The potential targets of SGSJD in treating PCM were obtained after the drug targets and disease targets were crossed. Cytoscape software was used to establish and analyze the network of Chinese medicines-active compounds-targets-disease; STRING database platform was used to analyze Protein Protein Interaction network; Bioconductor software package was used to perform Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment for potential targets. Western blot analysis was used to verify the janus kinase/signal transducer and activator of transcription (JAK-STAT) pathway . (a) 47 potential pharmacological components of SGSJD treatment of PCM were screened including quercetin, luteolin, kaempferol and others; 20 common targets were obtained, including interleukin-6 (IL-6), epidermal growth factor receptor, estrogen receptor 1, nitric oxide synthase 3 and others; a number of signal pathways were available, of which advanced glycation end product/ receptor for advanced glycation end products signaling pathway in diabetic complications, hypoxia-inducible factor 1 signaling pathway and janus tyrosine kinase-signal transducer and transcription activator (JAK-STAT) signaling pathway were the main signal pathways related to PCM. (b) Compared with the Blank group, the expressions of p-JAK2/JAK2, p-STAT3/STAT3 and IL-6 protein in the Model group were significantly increased ( < 0.01); Compared with the Model group, the expression of p-JAK2/JAK2, p-STAT3/STAT3, and IL-6 protein in the treatment group were significantly reduced in a dose-dependent manner ( < 0.05). Compared with the Model group, the dexamethasone significantly reduced the expression of p-JAK2/JAK2, p-STAT3/STAT3, and IL-6 ( < 0.01). The SGSJD may regulate the JAK-STAT signaling pathway to achieve the effect of treating PCM by reducing the expression of p-JAK2/JAK2, p-STAT3/STAT3 and IL-6 in a dose-dependent manner.

Identification of the active compounds and functional mechanisms of Jinshui Huanxian formula in pulmonary fibrosis by integrating serum pharmacochemistry with network pharmacology

BackgroundJinshui Huanxian formula (JHF), a traditional Chinese medicine (TCM), has been demonstrated to attenuate idiopathic pulmonary fibrosis (IPF). The active compounds and underlying mechanisms of JHF, however, are unclear. PurposeThe purpose of This study was to aimed to identify the active compounds and pharmacological mechanism of JHF by integrating serum pharmacochemistry with a network pharmacology strategy. MethodsJHF was orally administered to a rat model with bleomycin (BLM)-induced pulmonary fibrosis (PF). The pharmacodynamic effects and compounds present in the serum were identified. The targets and biological mechanisms of these compounds were revealed using network analysis and validated using in vitro experiments. ResultsJHF could significantly ameliorate BLM-induced PF by preventing extracellular matrix collagen deposition. Twenty-seven compounds that were found to be enriched in the serum samples collected 1 h after oral administration with JHF were identified as the candidate active compounds, and their 423 potential targets were identified as JHF targets. primarily related to the advanced glycation and products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, phosphatidylinositol 3 kinase (PI3K)-protein kinase B (PKB or AKT) signaling pathway, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance, etc. The 423 targets, 1145 IPF-related genes and their overlapped genes were applied to analyze, respectively. The results showed that these genes were primarily related to the advanced glycation end-products–receptor for advanced glycation end-products (AGE–RAGE) signaling pathway, lipid and atherosclerosis pathology, phosphatidylinositol 3 kinase (PI3K)-protein kinase B (PKB or AKT) signaling pathway, and epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance. Furthermore, the affinity between serum JHF compounds and the main proteins in the above important pathways was investigated through molecular docking. As a result, Molecular docking analysis showed that, tangeretin, isosinensetin, and peimine were found to could bind to EGFR and AKT, and their inhibitory effect on EGFR and AKT were validated in fibroblast cell induced by transforming growth factor (TGF)TGF-β. The results indicated that suppression of fibroblast activation by inhibiting the EGFR/PI3K/AKT signaling pathway might be an important mechanism of JHF may to treat PF. ConclusionJHF may suppress fibroblast activation by inhibiting the EGFR/PI3K/AKT signaling pathway to ameliorate PF. Tangeretin, isosinensetin, and peimine may be the active compounds in JHF involved in the treatment of that have therapeutic effects on IPF.

Using Network Pharmacology and Molecular Docking Technology to Explore the Mechanism of Modified Pulsatilla Decoction in the Treatment of Ulcerative Colitis

Objective: Using network pharmacology and molecular docking technology, our aim was to clarify the biological activity, key targets, and potential pharmacological mechanisms of modified Pulsatilla decoction (MPD) in the treatment of ulcerative colitis (UC). Materials and Methods: The main active ingredients of MPD were screened using the traditional Chinese medicine systems pharmacology platform. UC targets were obtained from the GeneCard, OMIM, DisGeNET, PharmGkb, and DrugBank databases. The common genes of MPD in the treatment of UC were identified by Venn diagram. The visual interactive network diagram of “active ingredient-target-disease” was constructed using the software Cytoscape. We used the STRING database to construct a protein–protein interaction network and analyze the correlation in protein interaction. We conducted gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis for common genes using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database and R software. Subsequently, the molecular docking verification of ingredients and targets was conducted through Discovery Studio. Last, in vivo experiments were conducted to further verify the findings. Results: A total of 51 active ingredients were screened, involving 141 common genes. The top 5 ingredients in MPD were quercetin, β-sitosterol, luteolin, kaempferol, and stigmasterol. Pathways involved in the treatment of UC include the advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, the interleukin-17 (IL-17) signaling pathway, the tumor necrosis factor (TNF) signaling pathway, viral infection-related signaling pathways, and some cancer pathways. Molecular docking showed that the important ingredients of MPD were well docked with mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase 8 (MAPK8), RAC-alpha serine (AKT1), vascular endothelial growth factor-A (VEGFA), transcription factor AP-1 (JUN), and interleukin-6 (IL-6). Animal experiments showed that MPD could ameliorate the injury and colitis in dextran sulfate sodium (DSS)-induced colitic rats. MPD inhibited the expression of p-p38A and p-MLC in UC rats. Conclusions: MPD has the characteristics of a multisystem, multi-ingredient, and multitarget in the treatment of UC. The possible mechanisms include inhibition of inflammation, apoptosis, oxidation, and tumor gene transcription. MPD may have a protective effect in the treatment of UC.

Contribution of the Microbiome to Western Diet‐Induced Gene Expression Changes in the Intestine and Vasculature

Western diet (high fat/sugar) is a core contributor to obesity, which is associated with increased risk for chronic diseases, including cardiovascular disease (CVD). The mechanisms through which Western diet contributes to CVD are heavily studied, and increasing evidence suggests that Western diet alters the gut microbiota leading to dysbiosis(i.e., disruption of microbial homeostasis and accompanying intestinal inflammation), contributing to development of vascular dysfunction, a strong predictor of future CVD. Still, the biological processes linking intestinal dysbiosis with the vasculature are incompletely understood. To identify potential mediators of intestinal‐vascular crosstalk, we investigated the role of the gut microbiome in Western diet‐induced global gene expression changes using transcriptomics. We performed total RNA sequencing on colons and arteries (aorta) from wild‐type (WT) and germ‐free (GF, no microbiome) mice fed standard or Western purified diets for 8 weeks. In the colon, we found ~100 differentially expressed genes in standard diet‐fed WT vs. GF mice, and pathway analyses showed that these genes were associated with several important biological processes including peroxisome proliferator‐activated receptor (PPAR) signaling, adipocyte lipolysis regulation, and AMP‐activated protein kinase (AMPK) signaling (genes and pathways, FDR &lt; 0.1). In colons of Western diet‐fed WT vs. GF mice, we found 163 differentially expressed genes related to cancer signaling, mitogen‐activated protein kinase (MAPK) signaling, and viral immune response (FDR &lt; 0.1). Interestingly, in transcriptome analyses of aortas we found significantly different expression of 314 genes in standard diet‐fed WT vs. GF mice, and these were related to complement/coagulation cascades, transforming growth factor beta (TGF‐β), and advanced glycation end product (AGE) signaling, but most of these differences were absent with Western diet (only 4 significantly different genes in WT vs. GF mice, FDR &lt; 0.1). These data suggest that the microbiome is an important mediator of gene expression in the intestine regardless of diet, but may only be important in arteries under standard diet conditions, and that the microbiome may not be a strong mediator of intestinal‐vascular crosstalk at the protein‐coding gene expression level. We are currently performing in‐depth analyses of non‐coding RNAs (e.g., micro‐RNAs) to further elucidate the mechanisms at play.

A Dual Anti-Inflammatory and Anti-Proliferative 3-Styrylchromone Derivative Synergistically Enhances the Anti-Cancer Effects of DNA-Damaging Agents on Colon Cancer Cells by Targeting HMGB1-RAGE-ERK1/2 Signaling.

The current anti-cancer treatments are not enough to eradicate tumors, and therefore, new modalities and strategies are still needed. Most tumors generate an inflammatory tumor microenvironment (TME) and maintain the niche for their development. Because of the critical role of inflammation via high-mobility group box 1 (HMGB1)–receptor for advanced glycation end-products (RAGE) signaling pathway in the TME, a novel compound possessing both anti-cancer and anti-inflammatory activities by suppressing the HMGB1-RAGE axis provides an effective strategy for cancer treatment. A recent work of our group found that some anti-cancer 3-styrylchromones have weak anti-inflammatory activities via the suppression of this axis. In this direction, we searched such anti-cancer molecules possessing potent anti-inflammatory activities and discovered 7-methoxy-3-hydroxy-styrylchromone (C6) having dual suppressive activities. Mechanism-of-action studies revealed that C6 inhibited the increased phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) under the stimulation of HMGB1-RAGE signaling and thereby suppressed cytokine production in macrophage-like RAW264.7 cells. On the other hand, in colorectal cancer HCT116 cells, C6 inhibited the activation of ERK1/2, cyclin-dependent kinase 1, and AKT, down-regulated the protein level of XIAP, and up-regulated pro-apoptotic Bax and caspase-3/7 expression. These alterations are suggested to be involved in the C6-induced suppression of cell cycle/proliferation and initiation of apoptosis in the cancer cells. More importantly, in cancer cells, the treatment of C6 potentiates the anti-cancer effects of DNA-damaging agents. Thus, C6 may be a promising lead for the generation of a novel class of cancer therapeutics.