Comparative analysis of melatonin and sildenafil in a rat model of pulmonary arterial hypertension: Insights into oxidative stress, inflammation, and mitochondrial biogenesis.

Clinical evidence suggests that essential hypertension is linked to oxidative stress and inflammatory infiltration, which can complicate osteoporosis. Edible house crickets (Acheta domesticus) are novel functional foods rich in proteins, fat, dietary fibers, and micronutrients. This research investigated the effects of a partially defatted house cricket powder (PDCP) on bone microarchitecture and strength in the spontaneously hypertensive rat (SHR) model. Fourteen female SHRs were divided into the experimental group receiving 300 mg/kg/day PDCP and the control group receiving the vehicle for 4 weeks. Blood pressure was determined by the CODA® non-invasive blood pressure system. Femoral bone microarchitecture and strength were determined by micro-computed tomography (µCT) and three-point bending, respectively. Immune cells were counted using an automated machine. Results showed that in the control group, systolic blood pressure (SBP) at baseline (166.6 ± 3.7 mmHg) increased to 182.4 ± 4.7 mmHg (P = 0.016). In contrast, the PDCP-treated group showed no significant change from baseline (168.1 ± 3.9 mmHg) to post-intervention (176.9 ± 5.6 mmHg) (P = 0.156). PDCP had no effects on bone microarchitecture but improved bone strength. The post-intervention yield load (a proxy for strength) of the control group was 77.55 ± 1.88 N, compared to the PDCP-treated group of 83.58 ± 1.26 N (P < 0.009). Similarly, post-intervention yield displacement was 307.72 ± 29.78 in the control vs. 395.93 ± 40.98 µm in the PDCP-treated group. White blood cell counts in the PDCP-treated group (7.91 ± 0.27 × 103/µL) were significantly higher than those in the control (6.91 ± 0.31 × 103/µL). Specifically, lymphocyte counts in the PDCP-treated group (6.66 ± 0.22 × 103/µL) were significantly higher than those in the control (5.75 ± 0.28 × 103/µL). In conclusion, the 4-week PDCP had osteoprotective effects, presumably mediated by dietary proteins and fibers that modulate immune-vascular homeostasis, thereby potentially mitigating immune system-related hypertension and bone mechanical impairment.

Sphingosine-1-phosphate (S1P), a metabolite of sphingosine, is associated with the proliferation of pulmonary artery smooth muscle cells (PASMCs). This study aims to address the mechanisms by which S1P induces PASMC proliferation, contributing to pulmonary arterial remodeling. Primary cultured rat PASMCs were incubated with S1P. Cyclopamine was used to inhibit Smoothened (SMO) function, while siRNA transfection selectively knocked down the expression of signal transducer and activator of transcription 3 ( STAT3 ), glioma-associated oncogene homolog 1 ( GLI1 ), and forkhead box M1 ( FOXM1 ). Cell proliferation was measured by 5-bromo-2'-deoxyuridine (BrdU) and 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. Subcellular localization of Gli1 was determined using immunofluorescence staining. In a monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) rat model, PF543 (the inhibitor of S1P synthetase), NSC74859 (the inhibitor of STAT3), and cyclopamine (the inhibitor of sonic hedgehog [Shh] receptor) were administered to evaluate their effects on disease progression. Hemodynamic changes and histological examination were performed to evaluate the development of PAH. The protein levels of sphingosine kinase 1 (SphK1), phosphorylated/total STAT3 (p-/t-STAT3), Shh, Gli1 and FoxM1 were determined using immunoblotting. S1P increased Shh expression by STAT3 activation, which further caused Gli1 upregulation and nuclear translocation in PASMCs. Activation of Gli1 raised FoxM1 expression and then triggered PASMCs proliferation and migration. In MCT-induced PAH rat models, S1P levels were elevated in lung tissues and serum, triggering STAT3 activation and subsequent upregulation of Shh, Gli1, and FoxM1 in lung. Targeting these molecules alleviated pulmonary arterial remodeling and prevented the development of PAH. S1P/STAT3/Shh/Gli1/FoxM1 pathway plays an important role in PASMCs proliferation and pulmonary arterial remodeling. Targeting this cascade may have potential value for the management of PAH.

The contribution of obesity to pulmonary arterial hypertension (PAH) pathophysiology remains poorly understood. Adipose tissue synthesises estrogens via cytochrome P450 (CYP) 19A1 (aromatase), whereas circulating estrogens are metabolised in the lung by CYP1A1. This study investigated whether obesity predisposes to PAH through enhanced estrogen synthesis and metabolism. A normoxic, two-hit, rat model of obesity-associated pulmonary hypertension (PH) was developed, combining Sugen 5416 (Sugen, Su) with a high-fat diet (HFD). Estrogen levels in SuHFD rat plasma and epicardial adipose tissue (EAT) from PAH patients were quantified using LC-MS/MS. CYP1A1 expression was assessed in lung and cardiac adipose tissue from SuHFD rats and PAH patients. The therapeutic potential of the CYP1A1 inhibitor hesperetin was evaluated in vivo. Complementary studies used pulmonary artery smooth muscle cells (PASMCs) from PAH patients and Simpson-Golabi-Behmel syndrome (SGBS) adipocytes. HFD-fed rats of both sexes developed mild PH, which Sugen moderately exacerbated. EAT from PAH patients exhibited up-regulated aromatase and CYP1A1 expression, along with elevated estrogen levels. Circulating estrone was increased in male SuHFD rats. Pulmonary CYP1A1 expression was elevated in SuHFD rats and PAH patients. Hesperetin attenuated obesity-associated PH, reducing CYP1A1 expression in SuHFD rat lungs and PAH PASMCs. CYP1A1 induction in female SuHFD rat pericardial adipose tissue and Sugen-treated SGBS adipocytes was also tempered. These findings implicate augmented estrogen production by adipose tissue and elevated pulmonary CYP1A1 expression in the pathogenesis of obesity-associated PH. CYP1A1 may represent a novel therapeutic target in obese PAH patients.

Long-acting formulations are increasingly sought after for the management of chronic diseases. Drug-conjugated microparticles with degradation-controlled release present a promising modular extended-release platform that can be delivered through minimally invasive approaches. However, many conventional drug-conjugated microparticles have relatively short durations of release and substantial release of pharmacologically inactive drug-conjugated oligomers. Here, we present surface-erodible, drug-conjugated polyketal microparticles for extended release of unmodified pharmacologically active drug. Microparticles made of a polyketal conjugated to the anti-glaucoma agent tafluprost initially release unmodified tafluprost, then about two weeks later predominantly release the biologically active tafluprost acid due to the mildly base-catalyzed hydrolysis of tafluprost, with a cumulative total drug release of ~78% over 540 days in vitro. Notably, no detectable drug-conjugated oligomers are found in release medium. Tafluprost-polyketal conjugate microparticles that further encapsulate unconjugated tafluprost reduce intraocular pressure for approximately 3 months after subconjunctival injection in ocular normotensive female rats, exhibiting a low grade of tissue reaction, no retinal abnormalities or systemic side effects, and enhanced intraocular pressure-lowering efficacy compared with a long-acting bimatoprost-loaded biodegradable implant in an ocular hypertensive rat model. Our findings have broad implications in advancing extended drug delivery systems.

Pulmonary hypertension (PH) is a progressive disease marked by pulmonary arterial remodeling and right ventricular dysfunction. The molecular mechanisms driving this remodeling, particularly ECM (extracellular matrix)-mediated processes, remain poorly understood. This study investigates THBS4 (thrombospondin-4), an ECM glycoprotein, as a key regulator of pulmonary vascular remodeling in pulmonary hypertension. Whole-transcriptomic analysis was conducted on pulmonary arteries from rat models of pulmonary hypertension induced by hypoxia, hypoxia-SUGEN, and monocrotaline. THBS4 expression was measured in these models and in serum and lung tissue from patients with pulmonary arterial hypertension. The role of HIF-1α (hypoxia-inducible factor 1-alpha), SMAD2, and p38 MAPK (mitogen-activated protein kinase) signaling pathways in regulating THBS4 was explored. Functional assays assessed THBS4's impact on pulmonary artery smooth muscle cells. In vivo, THBS4 silencing was performed to evaluate its effect on vascular remodeling and right ventricular hypertrophy. THBS4 was upregulated in pulmonary arteries across all pulmonary hypertension models, with expression correlating with disease severity. Elevated THBS4 levels were observed in pulmonary arterial hypertension patient serum and lung tissue. Hypoxia-induced THBS4 expression was mediated by HIF-1α, while TGF (transforming growth factor)-β1 stimulation enhanced THBS4 via SMAD2 and p38 MAPK pathways. THBS4 promoted pulmonary artery smooth muscle cell proliferation, phenotypic modulation, and ECM remodeling. In vivo silencing of THBS4 attenuated pulmonary vascular remodeling and right ventricular hypertrophy. Our findings identify THBS4 as a central regulator in a self-reinforcing THBS4-TGF-β/SMAD2 signaling axis driving pulmonary vascular remodeling. Targeting THBS4 represents a promising therapeutic strategy to mitigate pathological vascular remodeling in pulmonary hypertension.

Dysfunction of pulmonary artery endothelial cells (PAECs) contributes to the pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH). However, the role of mitochondrial metabolism in this process remains unclear. The present study evaluated whether the tetrameric form of pyruvate kinase muscle isoform 2 (PKM2) regulates PAEC mitochondrial metabolism through peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1alpha) and mitochondrial transcription factor A (mtTFA), thereby influencing arterial intimal remodeling in CTEPH. A CTEPH rat model was established by repeated injections of autologous thrombi. Activation of PKM2 tetramer expression was achieved through synthetic pyruvate kinase M2 activator (TEPP-46) administration. Pulmonary artery pressure (PAP), thrombus pathology, and protein expression levels of PKM2, mtTFA, and PGC-1alpha were assessed. Plasma lactate concentrations and tumor necrosis factor alpha (TNF-alpha) levels were measured. Rats with CTEPH demonstrated thrombotic obstruction, elevated PAP, and reduced expression of the PKM2 tetramer, mtTFA, and PGC-1alpha. Treatment with TEPP-46 was associated with a reduction in thrombus burden, lower PAP, and restoration of mitochondrial protein expression, accompanied by decreased lactate concentrations and TNF-alpha levels. In the CTEPH rat model, increased inflammation and elevated lactate concentrations were observed, along with decreased expression of mtTFA and PGC-1alpha in the pulmonary artery intima, which is indicative of mitochondrial dysfunction. The PKM2 tetramer may play a role in modulating PAEC mitochondrial function, reducing pulmonary artery pressure, and improving pulmonary arterial intimal remodeling in CTEPH. Key words Chronic thromboembolic pulmonary hypertension " Lactic acid " Mitochondrial transcription factor A " Peroxisome proliferator-activated receptor gamma coactivator 1alpha " Pyruvate kinase muscle.

Dysfunction of pulmonary artery endothelial cells (PAECs) contributes to the pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH). However, the role of mitochondrial metabolism in this process remains unclear. The present study evaluated whether the tetrameric form of pyruvate kinase muscle isoform 2 (PKM2) regulates PAEC mitochondrial metabolism through peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) and mitochondrial transcription factor A (mtTFA), thereby influencing arterial intimal remodeling in CTEPH. A CTEPH rat model was established by repeated injections of autologous thrombi. Activation of PKM2 tetramer expression was achieved through synthetic pyruvate kinase M2 activator (TEPP-46) administration. Pulmonary artery pressure (PAP), thrombus pathology, and protein expression levels of PKM2, mtTFA, and PGC-1α were assessed. Plasma lactate concentrations and tumor necrosis factor alpha (TNF-α) levels were measured. Rats with CTEPH demonstrated thrombotic obstruction, elevated PAP, and reduced expression of the PKM2 tetramer, mtTFA, and PGC-1α. Treatment with TEPP-46 was associated with a reduction in thrombus burden, lower PAP, and restoration of mitochondrial protein expression, accompanied by decreased lactate concentrations and TNF-α levels. In the CTEPH rat model, increased inflammation and elevated lactate concentrations were observed, along with decreased expression of mtTFA and PGC-1α in the pulmonary artery intima, which is indicative of mitochondrial dysfunction. The PKM2 tetramer may play a role in modulating PAEC mitochondrial function, reducing pulmonary artery pressure, and improving pulmonary arterial intimal remodeling in CTEPH.

Angiotensin converting enzyme (ACE) regulates blood pressure via the renin-angiotensin and bradykinin systems. Though synthetic ACE inhibitors are more effective, they pose several side effects. Methyl palmitate (MP), a natural fatty acid methyl ester with cytoprotective, antioxidant, anti-inflammatory, and vasodilatory properties, is not explored for its ACE inhibitory or antihypertensive potential. This study aimed to investigate the in vitro ACE inhibition of MP and its effect on L-NG-Nitro Arginine Methyl Ester (<sc>L</sc>-NAME)-induced hypertensive male Wistar rats. An in vitro ACE inhibition assay was conducted to compare the inhibition potency of MP with that of lisinopril. Male Wistar rats (n = 35, 7 per group) were grouped into control, disease control, and treatment groups receiving 100, 150, or 200 mg/kg/day of MP for 21 days each. Blood pressure, serum ACE activity, malondialdehyde (MDA), and nitric oxide (NO) levels in kidney tissue homogenate, and thoracic aorta histopathology were assessed. MP inhibited ACE by 61.05% at 5 µ<sc>m</sc>, exceeding lisinopril's 41.67%. High-dose MP significantly reduced serum ACE and MDA levels, while increasing NO (p < 0.001). Histopathology revealed near-normal vasculature, although changes in blood pressure were not statistically significant (p > 0.05). MP demonstrates strong natural ACE inhibition, antioxidant, and vascular protective effects, supporting further research for therapeutic optimization.

HHcy promotes hypertension by activating ferroptosis via suppression of the renal H₂S-SLC7A11/GPX4 axis.

Benign prostatic hyperplasia (BPH) is a highly prevalent age-associated disorder and a leading cause of lower urinary tract symptoms in men worldwide. Given the limitations of current therapies, there is increasing interest in phytotherapeutic compounds as sources of biologically active agents. Acmella oleracea, a medicinal plant rich in the alkamide spilanthol, has been traditionally associated with urogenital effects; however, the biological impact of distinct plant organs on prostate hyperplasia remains poorly defined. In this study, we investigated the effects of flower (A.Fl) and leaf (A.Le) extracts of A. oleracea using human prostate cell lines (RWPE-1 and PC-3) and a spontaneously hypertensive rat (SHR) model of BPH. In vitro analyses included cell viability assays and immunofluorescence for androgen receptor (AR), estrogen receptor alpha (ERα), and proliferating cell nuclear antigen (PCNA). In vivo, SHR were treated orally with A.Fl or A.Le (100 mg/kg/day for 21 days), followed by morphological, immunohistochemical, ultrastructural, and oxidative stress analyses of the ventral prostate. A.Fl displayed lower cytotoxicity than A.Le in both prostate cell lines and preferentially increased ERα immunoreactivity, whereas A.Le more strongly modulated AR without affecting cell proliferation. In SHR, both extracts attenuated prostatic hyperplasia, although A.Fl produced a more pronounced reduction in epithelial proliferation and stromal remodeling. These effects occurred independently of changes in systemic blood pressure or antioxidant activity. Collectively, these findings demonstrate that flower and leaf extracts of A. oleracea exert distinct biological and endocrine-modulatory effects on prostate tissue. The present data provide experimental evidence that different plant organs differentially influence epithelial-stromal dynamics and steroid receptor signaling in prostatic hyperplasia, supporting further mechanistic and translational investigations.

Chronic thromboembolic pulmonary hypertension is characterized by proximal pulmonary artery obstruction and distal microvasculopathy. However, the mechanisms driving this dual-compartment pulmonary vascular remodeling remain unclear. Male Sprague-Dawley rats were injected with gelatin sponge combined with SU5416 as a secondary insult. Hemodynamics, echocardiography, and pulmonary vascular remodeling were evaluated to investigate the development of chronic thromboembolic pulmonary hypertension. Single-cell RNA sequencing of rat lung tissue was conducted to elucidate the molecular mechanisms underlying pulmonary vascular remodeling. The results were validated by immunofluorescence and cell-based experiments. The optimal size range of gelatin sponge for large pulmonary artery obstruction was 710 to 1000 µm, which synergized with a low dose of SU5416 (10 mg/kg) to induce significant increases in right ventricular systolic pressure and right ventricular hypertrophy at 5 weeks. The model exhibited persistent elastic pulmonary artery obstruction and remodeling, along with significant wall thickening and muscularization of pulmonary microvessels. Single-cell transcriptomic analysis revealed a significant reduction in microvascular endothelial cells and an increase in smooth muscle cells in the chronic thromboembolic pulmonary hypertension rats. STAT3, EGR1, and HIF1A were identified as key genes driving aberrant smooth muscle cell proliferation. The Sell (encoding L-selectin)-Podxl (encoding PODXL, podocalyxin) ligand-receptor pair was found specifically in diseased rats and mediated immune cell-endothelial cell interactions. L-selectin promoted neutrophil adhesion and dysfunction in pulmonary arterial and microvascular endothelial cells, both of which were reversed by PODXL knockdown. Our new model recapitulates human chronic thromboembolic pulmonary hypertension pathophysiology and is useful for understanding pulmonary microvasculopathy. Sell-Podxl is a previously unrecognized link between inflammation and vascular remodeling, offering a potential therapeutic target.

This study identifies corn silk extract (CSE) as a novel plant-derived prebiotic with antihypertensive effects mediated through gut microbiota modulation. Using a spontaneously hypertensive rat model, we demonstrated that CSE reshapes gut microbial composition, enhances microbial diversity, and promotes beneficial genera while reducing systemic inflammation and restoring nitric oxide (NO)-mediated vascular function. Importantly, fecal microbiota transplantation confirmed the causal role of gut microbiota in mediating these effects. These findings highlight a gut microbiota-inflammation-NO axis as a key pathway through which CSE regulates blood pressure. As a safe, accessible, and food-compatible intervention, CSE represents a promising strategy for non-pharmacological blood pressure management and broadens the application scope of prebiotics in cardiovascular health.

PurposeEmerging evidence implicates ferroptosis in trabecular meshwork (TM) injury during glaucoma pathogenesis. This study aims to systematically characterize ferroptosis-related gene signatures in primary open-angle glaucoma (POAG) and identify small-molecule therapeutics targeting this mechanism.MethodsWe performed Mendelian randomization (MR) analysis of East Asian genome-wide association study data to assess causal relationships between blood metals and POAG. Ferroptosis-related indicators (iron ions, MDA, GPX4, GSH) were measured in aqueous humor samples from patients with POAG using ELISA. Genome-wide expression of TM from patients with POAG was integrated with ferroptosis-related gene sets (FerrDb) to identify core regulatory genes. The CMap database and molecular docking were employed to screen therapeutic compounds, with subsequent validation in hydrogen peroxide (H2O2)–induced TM ferroptosis models. Digoxin was formulated as topical ophthalmic drops and administered twice daily to chronic ocular hypertension (COH) rat models to evaluate its IOP-lowering efficacy.ResultsMR analysis revealed that elevated serum iron levels causally increase POAG risk. Patients with POAG exhibited increased iron and MDA levels alongside decreased GPX4 and GSH. We identified 14 ferroptosis-related differentially expressed genes, with PPI analysis pinpointing SLC2A3, SCD, and HBA1 as hub genes linking ferroptosis to TM injury. Their dysregulation amplified lipid peroxidation cascades. CMap screening and molecular docking suggested ATPase inhibitor digoxin as a potential POAG therapeutic. Digoxin significantly attenuated H2O2-induced ferroptosis in HTMCs by reducing MDA and restoring GPX4, ultimately lowering IOP in COH eyes.ConclusionsOur findings establish digoxin as a promising therapeutic agent that suppresses ferroptosis and reduces IOP via the SLC2A3/SCD/HBA1 pathway, providing a transformative strategy for POAG treatment.

Glaucoma is a major cause of irreversible blindness, characterized by the progressive degeneration of retinal ganglion cells (RGCs), with oxidative stress and apoptosis playing central roles in its pathogenesis. Sirtuin 3 (SIRT3) has demonstrated antioxidant and anti-apoptotic effects in various neurodegenerative diseases; however, its precise role in glaucoma remains unclear. This study aimed to elucidate the neuroprotective function and mechanistic basis of the SIRT3/AMP-activated protein kinase (AMPK) axis in glaucoma. A rat model of chronic ocular hypertension (COH) was generated using cross-linked hydrogel injection, while an N-methyl-D-aspartate (NMDA)-induced RGC injury model was developed in vitro. SIRT3 overexpression was achieved using adeno-associated virus (AAV) transfection, either alone or combined with the AMPK inhibitor Compound C. Functional and molecular analyses were performed, including intraocular pressure (IOP) measurement, hematoxylin-eosin (H&E) staining, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, immunofluorescence, Cell Counting Kit-8 (CCK-8) cell viability assay, flow cytometry, Quantitative real-time PCR (qRT-PCR), and western blotting. In the COH model, both SIRT3 expression and the p-AMPK/AMPK ratio were significantly reduced at weeks 2, 4, and 6 (p < 0.05). Overexpression of SIRT3 lowered IOP, preserved retinal thickness, and decreased the number of TUNEL-positive cells (p < 0.001), while Compound C partially reversed these effects (p < 0.05). In addition, SIRT3 overexpression markedly educed reactive oxygen species (ROS) accumulation (p < 0.001) and restored the p-AMPK/AMPK ratio (p < 0.001), both of which were partially inhibited by Compound C. In NMDA-induced RGCs, SIRT3 overexpression significantly increased SIRT3 mRNA levels (p < 0.01), enhanced cell viability (p < 0.001), and suppressed apoptosis (p < 0.001), with these effects attenuated by Compound C (p < 0.01). The reduction of ROS and activation of AMPK by SIRT3 in this model were also partly reversed by AMPK inhibition (p < 0.01). This study provides the first comprehensive in vivo and in vitro evidence in glaucoma models that SIRT3 confers neuroprotection in experimental glaucoma, primarily through activation of the AMPK signaling pathway. These findings identify the SIRT3/AMPK axis as a novel mechanistic target and suggest a promising therapeutic strategy for IOP-independent neuroprotection in glaucoma.

Hypertension remains a major global health burden, yet the optimal exercise strategy to mitigate its associated inflammatory and oxidative stress responses is still unclear. This study aimed to compare the effects of high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) on markers of inflammation and oxidative stress, including serum IL-6, NOX4, and cardiac tissue MMP-2 in a rat model of hypertension. Hypertension was induced in male Wistar rats through L-NAME administration for five weeks, during which the rats concurrently underwent HIIT or MICT. L-NAME administration resulted in a progressive increase in systolic blood pressure (SBP) by week five compared to control rats. L-NAME significantly increased serum IL-6 and NOX4 levels but did not significantly affect cardiac MMP-2 expression. MICT significantly reduced SBP and NOX4 levels, whereas IL-6 and MMP-2 levels remained unchanged. In contrast, HIIT did not produce significant reductions in SBP, IL-6, NOX4, or MMP-2. Statistical significance was defined as p ≤ 0.05. In conclusion, these findings indicate that MICT is more effective than HIIT in reducing elevated SBP and NOX4 levels in L-NAME-induced hypertensive rats.

Erectile dysfunction (ED) is widespread among individuals with high blood pressure and negatively affects quality of life. The effect of stromal vascular fraction (SVF) on hypertension-related ED remains unexplored. We used a hypertensive rat model to explore the relative efficacy of adipose tissue stromal vascular fraction (tSVF) and cellular SVF (cSVF). We then investigated the possible mechanisms of these treatments. Hypertensive rats were divided into three groups according to different treatments. Their intracavernous pressure (ICP) during erection and condition of cavernous tissue were compared to those of the controlled group. Endothelial-mesenchymal transformation (EndMT) markers as well as related inflammatory factors were also measured. cSVF and tSVF were labeled with CM-Dil before injection in order to determine whether cSVF and tSVF survived, proliferated, and transdifferentiated in vivo. The increased ICP during erection demonstrated that tSVF treatment significantly improved hypertension-related ED. tSVF increased the smooth muscle-to-collagen ratio and inhibiting the expression of fibrosis-related proteins in hypertensive rats while rescuing the expression of vWF and eNOS, which indicated the preserving of endothelial tissue of the penis. Immunofluorescence staining and western blotting of penile tissue clearly suggest the inhibitory effect of tSVF on the overoccurring EndMT. Immunofluorescence staining and Western blot analysis of endothelial cells in vitro corroborate the whole-tissue findings. The experiments in N-nitro-L-arginine methyl ester hydrochloride (L-NAME)-induced human umbilical vein endothelial cells (HUVECs) revealed tSVF suppresses EndMT via inhibiting the TGF-β2-Smad2/Smad3 pathway. In vivo tSVF and cSVF tracing suggested that tSVF showed better longevity and transdifferentiation capacity than cSVF, thus exerting a more significant therapeutic effect. Treatment with tSVF significantly reserved erectile function in a hypertensive rat model. The mechanism appears to be inhibition of pathological EndMT through self-differentiation. We conclude that tSVF is a promising therapeutic candidate for treating hypertensive ED.

This study evaluates the protective effects of kaempferol on right ventricular (RV) structure and function in a rat model of high-altitude pulmonary hypertension (HAPH) and explores the underlying mechanisms. Male Sprague-Dawley rats (n = 57) were exposed to a simulated high-altitude environment (5000 m) for 28 days to induce HAPH, followed by the administration of kaempferol (25, 50, 100 mg kg-1 day-1) or sildenafil (30 mg kg-1 day-1). Echocardiography, hemodynamic measurements, and histopathological analyses revealed that kaempferol significantly reduced the RV/(LV + S) index, mean pulmonary artery pressure (mPAP), mean right ventricular systolic pressure (mRVSP), and RV free wall thickness (RVFWT); increased the pulmonary artery acceleration time (PAAT), PAAT/pulmonary artery ejection time (PAET) ratio, tricuspid annular plane systolic excursion (TAPSE), and RV-pulmonary artery coupling indices (TAPSE/mPAP and TAPSE/pulmonary artery systolic pressure [PASP]); and attenuated RV remodeling (assessed by H&E staining, Masson's trichrome staining, and transmission electron microscopy). Phosphoproteomic screening identified AMPK signaling as a key pathway modulated by kaempferol. In vivo, kaempferol downregulated the hypertrophy marker (MYH7B), suppressed the AMPK/ACC/CPT1B axis (at 25/50 mg kg-1), and reduced the LDHA (at 50/100 mg kg-1) and p-PDHA1 levels (at 25/50 mg kg-1) in RV tissues. Using the CCK-8 assay, fluorescence microscopy, and hypertrophy marker (BNP/MYH7B) detection, the optimal concentrations of Ang II (1 × 10-6 M) and kaempferol (40 μM) were selected. The effects of kaempferol on AMPK-mediated glucose and fatty acid oxidation were partially reversed by AICAR co-treatment, confirming the involvement of the AMPK pathway. Collectively, these findings demonstrate that kaempferol protects the RV structure and function and attenuates RV remodeling in HAPH rats, potentially through the modulation of the AMPK/ACC/CPT1B axis and the AMPK-mediated LDHA/PDHA1 pathway.

We investigated the retinal response to acute elevation of intraocular pressure (IOP) in rat eyes. IOP elevation was induced in the right eyes of adult Wistar rats using laser photocoagulation of the trabecular meshwork, while the left eye served as untreated controls. Retinal layer thickness was measured in vivo using a home-built spectral-domain optical coherence tomography (SD-OCT) system. Glial cell reactivity was assessed using immunofluorescence staining for glial fibrillary acidic protein (GFAP). IOP was significantly increased in treated eyes one day after laser treatment (31.4±6.4mmHg), returning to normal in most animals by the second day (10.1±2.0mmHg). We followed the retinal layer thickness for 4 weeks (n=14) with a subset for 9 weeks (n=5). OCT images showed a significant increase in retinal layer thickness that persisted for 9 weeks. Histological analysis of retinal sections corroborated the in vivo OCT findings. Furthermore, the immuno-biochemical analysis revealed a marked GFAP upregulation, primarily in the retinal nerve fiber layer (RNFL). Quantification of retinal ganglion cells (RGCs) showed no significant loss of RGCs in the treated eyes in ten days after laser photocoagulation. These findings demonstrated that acute IOP elevation in this rat model induces a persisted significant increase in retinal layer thickness, rather than the commonly expected decrease in RNFL thickness. This thickening is associated with the activation of glial cells, suggesting a potential mechanism for this response.

The study also emphasizes the deployment of economical synthetic methods for obtaining more potent derivatives of commercially available drugs. The present study aimed to synthesize schiff-base (SB) derivatives of atenolol (ATN) and their metal complexes with copper and zinc; further characterized by various spectroscopic techniques as well as biological assay. The synthesized compounds are subjected to in-silico docking and ADME (using Swiss ADME program) studies to confirm their suitability as lead molecule. Antioxidant, antimicrobial and anticancer activities were carried out using DDPH, disk diffusion method and MTT using HepG2 cell lines respectively. Additionally; Anti-hypertensive activity was tested L-NAME induced hypertensive rat model through invasive method. Molecular docking studies indicate higher score for 4c and 5e than parent drug atenolol (ATN); SB and their metal derivatives also indicate promising therapeutic profile with good GIT absorption parameters. Here 4a represents potent antioxidant activity with IC50 = 8.95 ± 11μg/ml; when compared to std. drug Ascorbic acid with an IC50= 8.3± 11μg/ml. Furthermore; all the metal complexes are reactive against B.Subtilis a gram +ve bacteria than gram -ve E.Coli, 4a (39.31mm) and 4b (42.02mm) possessed higher antimicrobial activity when compared to std. drug Vancomycin (31.48mm). The results from MTT assay reveals higher anticancer potential (%age inhibition) of 4a (73%), 4d (75%) and 5d (74%) when compared to std. drug Doxorubicin (101.2%) at HepG2 cell lines. In-vitro analysis shows significant results with SB metal derivatives, where, Zn complexes of SB derivatives (5a-e) represents excellent anti-hypertensive activity in rats in contrast to standard drug alone. The imine Schiff based derivatives of atenolol and their metal complexes exhibit good biological efficacy as well as multi modal effects when compared to the frequently prescribed drug, atenolol.