Background:Neuroinflammation in the hypothalamic paraventricular nucleus (PVN) drives sympathetic overactivity in hypertension. The Anjiang Formula (AJ) shows clinical antihypertensive potential; however, the precise molecular targets mediating its central neuroprotective effects remain undefined. Methods:In this translational study, we investigated the clinical efficacy of AJ and tested the hypothesis that it directly inhibits the central RhoA/ROCK2 signaling axis. We integrated a retrospective cohort analysis with mechanistic validation. Clinically, 85 elderly patients with Grade 1 essential hypertension were treated with AJ (n = 43) or lifestyle control (n = 42) for 8 weeks. Target engagement was verified using surface plasmon resonance (SPR), microscale thermophoresis (MST), and cellular thermal shift assays (CETSA). Mechanisms were validated in Spontaneously Hypertensive Rats (SHRs) and Angiotensin II-stimulated microglia. Results:Clinically, AJ reduced systolic blood pressure (SBP) by a mean difference of 10.2 mmHg compared to controls (p < 0.001), with a 93% responder rate. This was accompanied by improved flow-mediated dilation (+1.3%) and reduced serum IL-6. Biophysical assays identified Shinflavanone as a direct ROCK2 ligand (K D = 20.0 nM; CETSA ΔT m = +5.2 °C). In SHRs, AJ lowered blood pressure and suppressed PVN microglial activation. In vitro, AJ inhibited the RhoA/ROCK2 cascade, downregulated JUN, and upregulated CREB1/NQO1, thereby reducing oxidative stress. These effects were abolished by the ROCK2 agonist lysophosphatidic acid. Conclusion:AJ provides antihypertensive efficacy in elderly patients. These benefits are mechanistically driven by Shinflavanone-mediated inhibition of ROCK2, which attenuates central neuroinflammation and restores redox homeostasis in the PVN.

Hypertension caused cardiomyocyte apoptosis and remodeling, leading to heart failure. Wogonoside (WOG), a flavonoid from Qingda granules, was evaluated for cardioprotective effects and mechanisms. Spontaneously hypertensive rats (SHRs) received WOG (0.075, 0.75, or 7.5 mg/kg/d) or valsartan (7.5 mg/kg/d) for 10 weeks. Blood pressure, cardiac function, histology, fibrosis, cardiomyocyte size, and apoptosis were assessed using echocardiography, hematoxylin and eosin, Masson, TUNEL and wheat germ agglutinin staining. Network pharmacology and Kyoto Encyclopedia of Genes and Genomes analyses identified targets and pathways. In vitro, Annexin V/propidium iodide staining, JC-1 staining, and Western blotting were adopted for the assessment of apoptosis and mitochondrial function in H9C2 cells stimulated by angiotensin II (Ang II). WOG treatment reduced elevated blood pressure and increased left ventricular ejection fraction and left ventricular fractional shortening in SHRs. Network pharmacology analysis revealed 74 overlapping targets enriched in apoptosis and MAPK pathways. WOG reduced myocardial hypertrophy, fibrosis, and apoptosis in SHRs, as evidenced by decreased expression of cleaved caspase-3 and Bax, and increased expression of Bcl-2. WOG treatment reduced cell apoptosis and mitochondrial depolarization, downregulated Bax and Cleaved caspase 3, and upregulated Bcl-2. Mechanistically, WOG treatment suppressed the phosphorylation levels of ERK, p38MAPK, and JNK in Ang II-stimulated H9C2 cells, as evidenced by decreased ratios of p-ERK/ERK, p-p38MAPK/p38MAPK, and p-JNK/JNK, in Ang II-stimulated H9c2 cells. WOG relieves cardiac injury and cardiomyocyte apoptosis induced by hypertension, likely by suppressing multiple signaling pathways including the MAPK signaling pathway.

BackgroundBaicalin shows potent vasculoprotective effects against hypertension despite poor oral bioavailability. We investigated whether gut microbiota modulation contributes to the systemic vasculoprotective effects of orally administered baicalin.MethodsWe utilized an Angiotensin II-induced hypertensive mouse model, employing broad-spectrum antibiotics, 16S rRNA sequencing, metabolomics, and in vitro co-culture assays to map the gut-immune-vascular axis.ResultsOral Baicalin significantly attenuated Ang II-induced blood pressure elevation and improved the intestinal barrier integrity. Antibiotic-induced microbiota depletion substantially weakened these protective effects, supporting a major microbiota contribution under the present experimental conditions. Baicalin reshaped the gut microbial community, enriched SCFA-supporting taxa, and restored a putative butyrate-associated microbial signature, enriching Akkermansia and Lactobacillus accompanied by increased cecal butyrate levels. Exogenous sodium butyrate recapitulated several major protective features of Baicalin treatment, including expansion of Foxp3+ regulatory T cells in mesenteric lymph nodes. These immune changes were accompanied by increased Foxp3+ regulatory immune-cell accumulation in the aortic adventitia. In vitro, butyrate-licensed Tregs suppressed Ang II-induced vascular smooth muscle cell (VSMC) proliferative responses, at least partly through IL-10-mediated inhibition of MAPK/ERK signalling.ConclusionBaicalin alleviates Ang II-associated vascular remodelling, at least in part, by reprogramming gut microbial ecology, increasing luminal butyrate availability, promoting regulatory immune responses, and suppressing VSMC proliferative signalling.

OBJECTIVE:To identify candidate biomarkers of blood stasis syndrome (BSS) associated with coronary artery disease (CAD) and explore the underlying inflammatory mechanisms. METHODS:Using the Olink Target 96 Inflammation panel, we identified plasma proteins in a group of 88 patients comprised of healthy controls (HCs), those with CAD and BSS (CAD-BSS), those with CAD without BSS (CAD-non-BSS), and those with BSS without CAD (non-CAD-BSS) (n = 22 in each group). Protein molecules that were specifically expressed in CAD or BSS were identified by differential expression analyses. Subsequently, potential protein biomarkers were identified using least absolute shrinkage and selection operator regression to enable CAD and BSS differentiation. The potential functional mechanisms of identified proteins were then determined by Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses. RESULTS:Patients with CAD had 31/92 upregulated and 4/92 downregulated proteins compared with those without. Chemokine (C-C motif) ligand 11 (CCL11), CUB domain-containing protein 1, hepatocyte growth factor, sirtuin 2 (SIRT2), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), CCL25, and tumor necrosis factor (TNF) showed the strongest upregulation (all P <0.0001). Patients with BSS had 8/92 downregulated proteins, specifically CCL28, CCL11, cystatin D, STAM-binding protein, 4E-BP1, matrix metalloproteinase-10, SIRT2, and monocyte chemotactic protein 4, compared with those without (all P < 0.05). The CAD-BSS group had one interleukin-17 (IL-17) upregulated and 10/92 downregulated proteins compared with the CAD-non-BSS group. When compared with the non-CAD-BSS group, the CAD-BSS group had 8 upregulated proteins but only 2 downregulated proteins, namely interleukin-10 receptor subunit alpha (IL-10RA) and TNF-related activation-induced cytokine (both P < 0.05). Totally 10 proteins were identified as potential candidate biomarkers of BSS in CAD patients. After least absolute shrinkage and selection operator regression analysis, two proteins that distinguished between BSS and non-BSS individuals among CAD patients were identified (SIRT2 and 4E-BP1). These proteins are primarily associated with the mechanistic target of rapamycin signaling pathway, which regulates inflammation and oxidative stress. CONCLUSIONS:Results suggest that the inflammatory response and mechanistic target of rapamycin signaling pathway participate in CAD and BSS development, and that SIRT2 and 4E-BP1 are prospective protein biomarkers for patients with CAD and BSS.

Cardiac injury is a severe complication of 5-fluorouracil (5-FU) treatment in patients with gastrointestinal tumors, underscoring the urgent need for effective therapeutic strategies. Tetramethylpyrazine (TMP), a bioactive compound derived from traditional Chinese medicine, has demonstrated promising potential for alleviating 5-FU-induced cardiotoxicity. However, the precise mechanisms underlying its cardioprotective effects remain poorly understood. This study aimed to investigate the cardioprotective effects of TMP on 5-FU-induced cardiac injury and elucidate the underlying molecular mechanisms. TMP intervention significantly alleviated 5-FU-induced cardiac injury in both in vivo and in vitro models, as evidenced by improved cardiac function, reduced histological damage, and decreased levels of injury markers (creatine kinase MB (CKMB), cardiac Troponin I (cTn-I), N-terminal pro-B-type natriuretic peptide (NT-proBNP)). RNA sequencing revealed that TMP suppressed the activation of the p38 MAPK/JNK/ERK signaling pathway in cardiac tissue following 5-FU treatment. Further in vivo and in vitro experiments confirmed that TMP treatment reduced 5-FU-induced PANoptosis in cardiomyocytes and inhibited the activation of p38 MAPK/JNK/ERK signaling pathway. The protective effects of TMP on PANoptosis were reversed by pathway activators. In conclusion, TMP alleviates 5-FU-induced cardiac injury and reduces cardiomyocyte PANoptosis via suppressing the p38 MAPK/JNK/ERK signaling pathway. These findings suggest that TMP is a promising therapeutic candidate for managing chemotherapy-induced cardiotoxicity.

Three-Strain Probiotic Combination (Golden Bifid), a probiotic formulation composed of Bifidobacterium longum, Lactobacillus bulgaricus, and Streptococcus thermophilus, is widely used to modulate gut microbiota homeostasis and treat various gastrointestinal disorders. However, the specific molecular mechanisms underlying its therapeutic effects in slow transit constipation (STC) remain incompletely understood. In this study, we demonstrated that Golden Bifid alleviates loperamide-induced constipation by coordinately modulating host transcriptomic profiles, particularly the MAPK and serotonin signaling pathways, and restoring gut microbiota composition and diversity. These multi-omics findings provide novel mechanistic insights into the clinical efficacy of this probiotic combination, which have not been previously elucidated. Using a loperamide (LOP)-induced STC rat model, Golden Bifid was shown significantly increase defecation frequency, fecal water content, and intestinal motility, while improving the pathological damage of colonic tissues. It also elevated the protein expression of c-kit, 5-HT, 5-HT3R, and 5-HT4R in colonic tissue. RNA sequencing identified 1,998 differentially expressed transcripts in Golden Bifid group compared with the LOP group, with 899 upregulated and 1,099 downregulated. These transcripts were enriched in pathways, such as the mitogen-activated protein kinase (MAPK), tumor necrosis factor (TNF) and estrogen signaling pathway. Additionally, 16S rDNA sequencing demonstrated that the Golden Bifid partially restored gut microbiota structure, increased microbial diversity, and reversed the dysbiosis induced by LOP, notably reducing the abundance of Patescibacteria and modulating microbial taxa at both the phylum and genus levels to resemble the gut microbiota composition of the control group. These findings suggest that Golden Bifid alleviate STC by enhancing c-kit and 5-HT signaling, modulating the MAPK signaling pathway and pathway and restoring gut microbiota balance, offering promising therapeutic potential for STC treatment.

Hypertriglyceridemia (HTG) is a significant risk factor for cardiovascular disease, fatty liver, and acute pancreatitis, yet remains a therapeutic challenge due to limitations of current treatment options. To address this unmet clinical need, we screened a natural small-molecule library at an initial concentration of 100 μM to identify effective HTG therapeutic candidates using a CRISPR/Cas9-generated apolipoprotein C2 (apoc2) knockout zebrafish model that resembles human lipid metabolism disorders. Phenotype-based screening identified paeoniflorin (PAE) from 351 compounds as a potent triglyceride-lowering agent. Lipidomics analysis revealed PAE promoted triglyceride lipolysis by β-oxidation and lipophagy. Mechanistic studies demonstrated PAE upregulates peroxisome proliferator-activated receptor α (ppara) and lipoprotein receptor (ldlr) in apoc2 mutants. In oleic acid-induced Huh7 human hepatocytes, PAE reduces intracellular lipid droplet accumulation and significantly upregulated PPARA and LDLR expression, indicating enhanced hepatocellular uptake and oxidative catabolism of triglyceride-rich lipoproteins. Further investigation revealed that PAE upregulates the expression of hepatocyte nuclear factor 4 α (HNF4A), a key upstream transcription factor of PPARA. The HNF4A inhibitor BI-6015 completely abolished PAE's triglyceride-lowering effects, suggesting mediation through the HNF4A-PPARA-LDLR axis. These findings establish PAE as a promising therapeutic candidate for HTG through a novel mechanism targeting the HNF4A-PPARA-LDLR pathway. Our work not only identifies a potential lead compound for HTG treatment but also supports the zebrafish model as an effective platform for discovering drugs targeting hepatic lipid metabolic pathways.

Objective To investigate the potential therapeutic effects of trifolin on hypertension-induced renal injury,as well as the key targets and pathways involved.Methods The mRNA transcriptional profiles of peripheral blood clinical samples from hypertensive patients were analyzed using Gene Expression Omnibus(GEO),a high-throughput gene expression database.The network pharmacology method was employed to screen key targets of trifolin in treating hypertension-induced renal injury.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were conducted.NRK-52E cells,a rat renal proximal tubular cell line,were used to construct an angiotensin Ⅱ(Ang Ⅱ)-stimulated cell model.Flow cytometry was performed to assess cell apoptosis rates and Western blotting was performed to determine the expression levels of apoptosis-related proteins,including Bax,Bcl-2,cleaved caspase-3,and caspase-3,and the phosphorylation and total protein levels of the key MAPK pathway proteins,including ERK,p38 MAPK,and JNK.Results Analysis of the dataset GSE75360 revealed that,compared with healthy controls,3 331 genes were upregulated and 3 197 genes were downregulated in peripheral blood mononuclear cells of hypertensive patients.According to network pharmacology analysis,472 potential targets of trifolin were identified,including CASP3 and MAPK1.Protein-protein interaction network analysis showed that these targets were closely associated with apoptosis regulatory signaling pathways.GO and KEGG pathway enrichment analyses indicated that trifolin was significantly enriched in pathways associated with negative regulation of apoptosis,apoptotic signaling pathways,and the MAPK signaling pathway.The in vitro experiments confirmed that,compared with the Ang Ⅱ group,trifolin intervention inhibited apoptosis in Ang Ⅱ-stimulated NRK-52E cells,suppressed the expression of Bax and cleaved caspase-3,promoted Bcl-2 expression,and inhibited the phosphorylation of p38 MAPK,ERK,and JNK(P<0.05).Conclusion Trifolin may exert its protective effect against hypertension-induced renal injury by inhibiting Ang Ⅱ-induced NRK-52E cell apoptosis and regulating the MAPK signaling pathway,representing an important mechanism underlying its therapeutic action.

BACKGROUND:Panax notoginseng saponin (PNS) has shown potent activities in treating patients with atherosclerosis (AS), whereas its immunometabolic mechanism remained unknown. OBJECTIVES:To elucidate the characteristics of metabolomics of AS plaque and identify immunometabolic mechanisms of PNS in treating AS. METHODS:PNS components were characterized using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE). Spatial Metabolomics was performed to reveal characteristics of plaque from atherosclerosis patients. Then we verified different glycolysis and sphingolipid metabolites between chronic coronary disease (CCD) patients and health control (HC). In vivo and in vitro experiments were performed to study the relationship among hypoxia-inducible factor-1 α (Hif-1α), glycolysis, and macrophage polarization, as well as the protective role and underlying mechanism of PNS in AS. RESULTS:Among 8058 metabolites, 117 metabolites were greatly downregulated and 355 metabolites were remarkably upregulated in the plaque area. Sphingolipid metabolism ranked the top according to KEGG analysis, among which sphingosine-1-phosphate (S1P) and specifically sphingomyelin (SPH) were significantly reduced in CCD patients. Overexpression of Hif-1α could induce macrophage M1 polarization, enhance glycolysis, promote excessive UDP-glucose ceramide glucosyltransferase (UGCG) production, and reduce S1P, resulting in a proinflammatory response. Chemical characterization of PNS verified the presence of the principal saponins (R1, Rg1, Rb1, Rd, Re), which subsequently was shown to act as a novel Hif-1α inhibitor, exerting its anti-atherosclerotic effects through multiple pathways, specifically, downregulating key glycolytic regulators (PFKFB3, GLUT1, HK2) and suppressing downstream glycolysis; promoting the expression of MRC1/iNOS to inhibit M1 macrophage polarization; upregulating S1P to modulate sphingolipid metabolism, thereby alleviating AS. CONCLUSION:These findings emphasize the anti-AS effects of PNS through a novel immunometabolic mechanism, particularly its role as a Hif-1α inhibitor that disrupts such pathways driving AS progression.

BACKGROUND:Although neferine exhibits obvious therapeutic effects against hypertension, its effects on cardiac protection remain unknown. PURPOSE:This study aimed to investigate its potential cardioprotective effects and associated mechanisms. METHODS:Spontaneously hypertensive rats (SHRs) were randomly divided into four groups, namely SHR, SHR + Neferine-L (2.5 mg/kg/day), SHR + Neferine-M (5 mg/kg/day), and SHR + Neferine-H (10 mg/kg/day). Wistar Kyoto rats were used as control. Various concentrations of neferine or double distilled water were then administered intragastrically for 10 weeks. Thereafter, cardiac function, pathological changes, cell apoptosis, and reactive oxygen species (ROS) accumulation, as well as their underlying mechanisms, were evaluated in SHRs and/or hypoxia-induced H9c2 cells. RESULT:Neferine treatment significantly mitigated the decrease in left ventricular ejection fraction and fractional shortening and increase in left ventricular mass, end-systolic volume, and cardiac injury in SHRs. In SHR cardiac tissues, neferine treatment reversed 154 upregulated and 108 and downregulated transcripts. Pathway enrichment analysis found that multiple pathways were commonly enriched, including the apoptosis, PI3K-Akt, MAPK, and HIF-1 pathways. Consistently, neferine treatment significantly mitigated cardiomyocyte apoptosis, restored mitochondrial membrane depolarization, and reduced ROS accumulation. Mechanistically, neferine treatment significantly decreased the phosphorylation of ERK, p38 MAPK, and JNK; the Bax/Bcl-2 ratio; and the expression of HIF-1α, NADPH oxidase 4, and cleaved caspases-3 and -9 but increased the phosphorylation of PI3K and Akt and the expression of CD31. CONCLUSION:Neferine treatment effectively mitigated hypertensive cardiomyocyte apoptosis and attenuated the abnormal activation of multiple signaling pathways, including the PI3K/Akt, MAPK, and HIF-1 pathways.

BACKGROUND:Gastric cancer (GC), a leading cause of mortality globally, is a complex condition stemming from various factors including unhealthy lifestyle habits, genetic mutations expressed in a single nucleotide, environmental effects, and chronic Helicobacter pylori infections. The development and progression of GC are gradual over many years, with a lack of specific symptoms in early stages. This results in late diagnosis at advanced stages when the cancer is more aggressive, which in turn significantly impacts the prognosis and treatment options. SUMMARY:This review examines research focused on genetic mutations and mechanisms that contribute to the development and advancement of GC. We analyzed the literature on genetic mutations in GC from 1 January 2012 to 12 March 2025. These studies were derived from PubMed, Google Scholar, and the Web of Science. The following MeSH terms were used to explore these databases and to derive the relevant data: "gastric cancer" AND "stomach cancer" AND "genetic mutations" OR "genes" AND "onset" OR "development" AND "progression" OR "advancement." KEY MESSAGES:Our analysis revealed various genetic and molecular alterations in GC, including the overexpression of receptors and cancer-causing genes and the inactivation of tumor-suppressing genes. APC, ATM, BRCA1, BRCA2, CDH1, CDH2, MLH1, MSH2, MSH6, SMAD4, TP53, PIK3CA, and RHOA were linked to the onset and progression of GC, with CDH1 mutations emerging as the most prevalent. Identifying the genetic alterations underlying the onset and development of GC is essential, as it can enable prompt diagnosis and help establish effective preventive measures for this condition.

Carbohydrate kinases serve an oncogenic role in several types of cancer; however, the function of FGGY carbohydrate kinase domain containing (FGGY) in colorectal cancer (CRC) remains unknown. The present study investigated the function and possible molecular mechanisms of FGGY in CRC. The results showed that elevated levels of FGGY mRNA and protein were observed in CRC tissues, and a higher expression of FGGY was associated with advanced N stage and reduced overall survival time in patients with CRC. Silencing FGGY inhibited the viability of CRC cells by inducing cell cycle arrest and promoting apoptosis in vitro, thereby attenuating tumor growth in a xenograft mouse model. FGGY knockdown also enriched the senescence‑associated heterochromatin foci (SAHF) pathway and p53 pathway, as further confirmed by enhancing senescence‑associated β‑galactosidase (SA‑β‑gal) activity, with increased levels of SAHF‑associated proteins HP1γ and trimethylation of H3K9 (H3k9me3) in CRC cells, as well as upregulation of p53 and its downstream protein p21. Furthermore, p53 knockout rescued FGGY knockdown‑mediated reductions in cell viability, SA‑β‑gal activity, and the levels of HP1γ and H3k9me3 in CRC cells. These findings indicated that FGGY could act as a newly identified potential oncogene in CRC, partially through regulating the p53/p21 signaling pathway and altering cell senescence.

Background:Raised erosive gastritis (REG) is a chronic gastritis with a high risk of malignant transformation. Current treatments often result in high recurrence rates and complications. Jianpi Qinghua Sanyu Yin (JPQHSYY), a traditional Chinese medicine, shows promise in treating REG. However, the underlying molecular mechanisms remain unclear. This study aimed to investigate the potential mechanism of JPQHSYY's therapeutic effects on REG. Methods:RNA-seq was employed to systematically analyze mRNA, lncRNA, and miRNA profiles in gastric mucosal tissues from REG patients before and after JPQHSYY treatment. The pivotal lncRNA-miRNA and miRNA-mRNA networks were predicted from sequencing data and bioinformatic analysis, and the results were exported using Cytoscape software. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used for functional exploration. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to validate RNA-seq analysis results. CCK8, cell cycle, apoptosis and western blot were performed to detect the effects of miR-122-5p in GES-1 cells in vitro. Results:RNA-seq analysis revealed 576 differentially expressed lncRNAs (269 upregulated, 307 downregulated), 33 differentially expressed miRNAs (13 upregulated, 20 downregulated), and 1717 differentially expressed mRNAs (777 upregulated, 940 downregulated) in JPQHSYY-treated REG patients. GO and KEGG analyses highlighted key pathways, including the PI3K/AKT signaling pathway, involved in cell cycle and apoptosis regulation. The ceRNA network analysis suggested that JPQHSYY impacts the miRNA-lncRNA interactions. Validation experiments confirmed that JPQHSYY inhibits the PI3K/AKT pathway, reducing cell viability, colony formation, and promoting apoptosis in miR-122-5p transfected GES-1 cells. Conclusion:The therapeutic efficacy of JPQHSYY in treating REG might be mediated by the ceRNA-driven PI3K/AKT pathway signaling pathways, which is implicated in the proliferation of gastric mucosal epithelial cells. Furthermore, the investigation of miRNA-lncRNA networks could reveal more information on potential new mechanisms and targets for JPQHSYY in the management of REG.

Endothelial dysfunction (ED) plays a pivotal role in the pathogenesis of hypertension and its associated vascular complications. Qingda granule (QDG) exhibits significant antihypertensive properties and demonstrates therapeutic potential in ameliorating vascular dysfunction. This study aimed to explore QDG's role in alleviating endothelial injury in hypertension. An L-NAME (Nω-Nitro-L-arginine methyl ester)-induced hypertensive mouse model was used to evaluate the effects of QDG on blood pressure and endothelial function. Endothelial function was assessed through histological analysis, nitric oxide (NO) quantification, and vascular response measurements. To explore underlying mechanisms, network pharmacology was conducted using databases such as HERB, SwissTargetPrediction and STRING. Key pathways related to inflammation and cell adhesion were identified. Based on these findings, immunohistochemical staining was conducted to analyze the expression of phosphorylation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (p-NF-κB p65), NF-κB p65, intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α) in vascular tissues. QDG treatment significantly reduced blood pressure, increased NO levels, and enhanced endothelial nitric oxide synthase (eNOS) expression in L-NAME-induced hypertensive mice, indicating its potential to restore endothelial function. Experimental validation further confirmed that QDG markedly suppressed the expression of p-NF-κB p65, TNF-α, and ICAM-1 in vascular tissues. These results suggest that QDG alleviates hypertension-induced ED primarily by inhibiting inflammation and endothelial adhesion via the NF-κB signaling pathway. Overall, QDG presents a promising therapeutic candidate for managing hypertension and its vascular complications.

Trifolin, a bioactive component of the Qingda granule, has demonstrated significant antihypertensive potential; however, its precise mechanisms of action remain largely unknown. This study aimed to investigate the antihypertensive effects of trifolin and unravel its underlying molecular mechanisms. The influence of trifolin on vascular contraction and relaxation and its regulatory effects on ion channels were evaluated through a vascular tension experiment. Morphological changes in the aortic tissues of mice with angiotensin Ⅱ-induced hypertension and the expression profiles of contraction-associated proteins were analyzed via hematoxylin-eosin staining and immunohistochemistry. Additionally, trifolin’s impact on calcium ion dynamics and contraction-associated protein expression in angiotensin Ⅱ-activated vascular smooth muscle cells (VSMCs) was determined through calcium flux assays and western blot analyses. Trifolin treatment decreased the constriction of isolated abdominal aortic rings induced by norepinephrine, KCl, and angiotensin Ⅱ in an endothelium-independent manner and extracellular Ca2+ influx induced by these three substances and thapsigargin. Moreover, trifolin treatment significantly reduced the abdominal aortic wall thickness and downregulated the expression of store-operated channels channel proteins (STIM1 and ORAI1) and calcium signaling-related proteins (CaM, myosin light chain kinase, and p-MLC2) in the abdominal aorta of hypertensive mice and angiotensin Ⅱ-induced VSMCs. In conclusion, calcium signaling inhibition may underlie trifolin’s antihypertensive effects and its ability to ameliorate vascular function. These findings offer new therapeutic insights for hypertension treatment.

Trifolin exhibits anti-tumor activities; however, its effect on hypertension remains unknown. This study was performed to investigate trifolin’s potential therapeutic effects and underlying mechanisms of action on angiotensin II (Ang II)-induced hypertension in mice and Ang II stimulated A7R5 cells. Mice were randomly allocated into six groups: control, Ang II, Ang II + Trifolin (0.1 mg/kg), Ang II + Trifolin (1 mg/kg), Ang II + Trifolin (10 mg/kg), and Ang II + Valsartan (10 mg/kg). The hypertensive mouse model was constructed by infusing Ang II via a micro-osmotic pump (500 ng/kg/min), and trifolin, valsartan, or double distilled water was administered intragastrically once daily for 4 weeks. Blood pressure, vascular function, pathological morphology, and collagen deposition in Ang II infused mice and cell viability of Ang II stimulated A7R5 cells were assessed. A networking pharmacology analysis was performed to identify potential targets, pathways, and processes. These were verified by determining proliferating cell nuclear antigen (PCNA) expression, cell migration, collagen protein expression and related pathway activation in vivo and in vitro using masson, immunohistochemistry, cell counting Kit-8 assays, phalloidin staining, wound healing assays, and western-blotting. Different concentrations of trifolin effectively mitigated the rise in systolic blood pressure, diastolic blood pressure, mean arterial pressure, pulse wave velocity, abdominal aorta wall thickness, and collagen deposition of Ang II infused mice. Notably, higher concentrations of trifolin exhibited greater attenuation which was similar to the effects of valsartan (a positive control). Networking pharmacology analysis identified 105 common targets and various gene ontology processes. The Kyoto Encyclopedia of Genes and Genomes pathways analysis identified multiple enriched signaling pathways, including responses to wounding, phosphatidylinositol 3-kinase complex, oxidoreductase, PI3K/AKT, and FoxO signaling pathways. Consistently, trifolin treatment significantly down-regulated the expression of PCNA and the ratio of p-PI3K/PI3K and p-AKT/AKT in the abdominal aorta tissues. In vitro study indicated that trifolin consistently reduced the cell viability, down-regulated the expression of PCNA, collagen I and collagen III, and reduced the cell migration, as well as reduced the ratio of p-PI3K/PI3K and p-AKT/AKT (similar with the effect of PI3K inhibitor: LY294002) in Ang II stimulated A7R5 cells. Trifolin treatment attenuated the elevation of blood pressure, the proliferation and collagen deposition of VSMCs, and modulated multiple signaling pathways, including PI3K/Akt pathway. These results suggest that trifolin could be a potential therapeutic approach for treating hypertension.

Death-associated protein kinase 1 (DAPK1) is a tumor suppressor gene involved in apoptosis, autophagy, and tumor progression. However, its role in hypertension (HTN) remains largely unexplored and lacks systematic evaluation. We administered adeno-associated virus (AAV) harboring short hairpin RNA targeting DAPK1 or control short hairpin RNA to male spontaneously hypertensive rats (SHRs) and Wistar-Kyoto rats. Additionally, wildtype and DAPK1 knockout mice were infused with angiotensin II (Ang II) or saline for four weeks. Male C57BL/6 mice underwent a four-week Ang II infusion and were treated with TC-DAPK6, a selective DAPK1 inhibitor. We examined the abdominal aortas (AAs) of mice and rats for pathological changes, measured blood pressure (BP) and pulse wave velocity using noninvasive BP methods, ultrasound, and hematoxylin and eosin staining. The role of DAPK1 in early HTN was further assessed through immunofluorescence, ex vivo isometric constriction of the AA, RNA sequencing, Western blot, and immunohistochemistry. Our study demonstrated that the targeted inhibition of DAPK1 with AAV significantly ameliorated HTN in SHRs and reduced damage to the AAs and target organs, including the heart and kidneys. Meanwhile, DAPK1 knockout or inhibition in mice significantly ameliorates Ang II-induced HTN in mice, as well as reducing damage to the AAs and target organs, including the heart and kidneys. Mechanistically, DAPK1 inhibition prevents myosin light chain (MLC) phosphorylation at serine 19, reducing vasoconstriction and protecting against HTN. In conclusion, DAPK1 is involved in HTN pathogenesis by regulating the MLC pathway to mediate vascular constriction, highlighting potential as a therapeutic target for HTN.

Objective:To investigate the potential therapeutic effects, targets, and pathways of wogonoside in hypertension-induced renal injury using the Gene Expression Omnibus (GEO) database and network pharmacology, and to validate the effects of wogonoside intervention on the renal tissues of spontaneously hypertensive rats (SHR), angiotensin Ⅱ (Ang Ⅱ)-stimulated NRK-52E cell apoptosis, and the regulation of relevant pathways through in vivo and in vitro experiments. Methods:GEO dataset and network pharmacology analyses were performed to investigate the key therapeutic targets of wogonoside for hypertensive nephropathy. The STRING database was used to analyze protein-protein interactions. Biological functions were annotated via Gene Ontology (GO), and the potential signaling pathways were enriched using the Kyoto Encyclopedia of Genes and Genomes (KEGG). SHR were randomly divided into groups and given low, medium, or high doses of wogonoside (0.075, 0.75, and 7.5 mg/kg) via gastric gavage for 10 weeks. Morphological changes in the kidney tissue were assessed by hematoxylin-eosin (HE) staining. Serum levels of inflammatory cytokines, including tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6, were measured using ELISA. Apoptosis rates were evaluated by TUNEL staining, and Western blot was performed to determine the expression of Bax, Bcl-2, cleaved caspase-3, and caspase-3, and the expression of phosphorylated and total extracellular signal-regulated kinases (ERK) and p38 mitogen-activated protein kinase (MAPK) proteins. An in vitro model of Ang Ⅱ-stimulated NRK-52E cells was constructed and was treated with wogonoside at different concentrations (25, 50, or 100 μmol/L) for 24 h. The apoptosis rates were then assessed by Annexin V staining, and Western blot was performed to validate the expression of apoptosis-related and pathway-associated proteins. Results:Analysis of dataset GSE41453 revealed 11673 upregulated and 5902 downregulated genes in the renal tissues of SHR compared to the Wistar Kyoto (WKY) rats, or the WKY control group. Through the analysis of multiple databases, 371 potential targets of wogonoside were identified, resulting in 98 overlapping targets. From these, 45 core therapeutic targets were identified through further analysis, including TNF, CASP3, etc. GO analysis significantly enriched processes such as the negative regulation of apoptosis. KEGG pathway enrichment analysis highlighted the apoptosis pathway, IL-17 signaling pathway, and MAPK signaling pathway as being significantly enriched. Wogonoside treatment effectively mitigated pathological damage in SHR kidney tissues and significantly inhibited the expression of inflammatory cytokines, including TNF-α, IL-1β, and IL-6 (P < 0.05). It also decreased cell apoptosis rates in SHR kidney tissues and Ang Ⅱ-stimulated NRK-52E cells, downregulated the expression of Bax and cleaved caspase-3, and upregulated Bcl-2 expression (P < 0.05). Furthermore, wogonoside treatment inhibited the phosphorylation of ERK and p38 MAPK in SHR kidney tissues and Ang Ⅱ-stimulated NRK-52E cells (P < 0.05). Conclusion:Wogonoside may exert its protective effects against hypertension-induced renal injury by suppressing the inflammatory response and cell apoptosis, potentially through the regulation of the MAPK signaling pathway.