U. Saeed
36612315100
Publications - 18
Bacterial infections and antimicrobial resistance patterns: a comprehensive analysis of health dynamics across regions in Pakistan (2013-2023)
Publication Name: Brazilian Journal of Biology
Publication Date: 2025-01-01
Volume: 85
Issue: Unknown
Page Range: Unknown
Description:
Antimicrobial resistance (AMR) is a significant public health concern globally, and Pakistan is no exception. The misuse and overuse of antibiotics, inadequate regulation of their sale, and a lack of awareness contribute to the rising levels of AMR in the country. study presents a detailed analysis of blood and urine samples collected in Pakistan over various periods, focusing on pathogen prevalence, gender distribution, and age-wise patterns. From January 2013 to 2017, the North region exclusively contributed to the blood sample dataset, with Salmonella emerging as the primary pathogen, particularly affecting infants and neonates. Subsequently, from January 2017 to December 2020, a significant dataset emerged from the North and Punjab regions, with Salmonella and E.coli prevalent across all age groups, notably impacting adults and infants. In the period from January 2021 to the present, blood samples predominantly originated from the North and Punjab regions, with Salmonella and E.coli remaining significant pathogens, affecting adults and the elderly. Regarding urine samples, from January 2013 to December 2017, E.coli was the dominant pathogen, with females showing a higher susceptibility to urinary tract infections (UTIs), particularly among the elderly. Similarly, from January 2017 to December 2020, E.coli remained predominant, with UTIs more prevalent in females and the elderly. In the most recent period, the North region significantly contributed to UTI cases, with E.coli remaining predominant and females exhibiting a higher susceptibility, especially among the elderly. This comprehensive analysis provides crucial insights into the epidemiology of blood and urinary tract infections in Pakistan, informing public health strategies and interventions aimed at addressing these health challenges.
Open Access: Yes
Lymphopenia as a diagnostic biomarker in clinical COVID-19: insights from a comprehensive study on SARS-CoV-2 variants
Publication Name: Brazilian Journal of Biology
Publication Date: 2025-01-01
Volume: 85
Issue: Unknown
Page Range: Unknown
Description:
The enduring SARS-CoV-2 pandemic necessitates robust tools for severity assessment. This study, conducted at Islamabad Diagnostic Center across Pakistan from January 2021 to August 2022, aimed to investigate hematological abnormalities among suspected SARS-CoV-2 subjects. Initial enrollment included 130,347 cases, with 53,078 confirmed positive and 77,269 negative. An additional 11,786 samples expanded the dataset to 142,133. The Omicron and Centaurus variants, in confirmed positive patients, exhibited a slightly higher frequency of hematological abnormalities (30.42%) than negative participants (27.01%). Notably, lymphocyte count reduction (40.95%) suggested its potential as an alternative diagnostic parameter for clinical COVID-19. Decreased levels of NA (37.99%), HGB (26.17%), MCV (20.60%), PLT (6.15%), and ALB (2.28%) were observed. Abnormally elevated NEU, CR, MONO, RBCs, WBC, and EOS levels affected 26.00%, 24.28%, 30.79%, 22.02%, 6.28%, and 5.53% of subjects, respectively. Comparatively, positive patients exhibited higher abnormal blood parameters—LYMP count (57.40%), NEU count (46.08%), EOS count (62.48%), MONO count (31.61%), RBC count (30.32%), ALC count (43.60%), CR count (30.91%), NA count (40.53%), CRP count (68.46%), and DD (63.08%) than negative counterparts. The study underscores lymphocytopenia’s potential as a cost-effective, early diagnostic biomarker for clinical COVID-19, preceding real-time PCR diagnosis. This supports its consideration in resource-limited settings for strategic screening and policy-making in the ongoing SARS-CoV-2 battle.
Open Access: Yes
Estimating high mobility group box protein 1 (HMGB1) single nucleotide polymorphisms among hepatitis B virus infected patients of Pakistan origin
Publication Name: Brazilian Journal of Biology
Publication Date: 2025-01-01
Volume: 85
Issue: Unknown
Page Range: Unknown
Description:
HMGB1 is nuclear non-histone protein and unique member of cytokines. In viral hepatitis infection HMGB1 serum level increases and translocates towards cytoplasm and extracellular spaces where it activates single stimulating hepatic stellate cell proliferation which induces fibrogenic protein expression and causes hepatocellular carcinoma. In this study, total 150 subjects were recruited to assess the association between HMGB1 SNPs and HBV. Three types of genotypes were found visible in rs3742305 of HMGB1; wild type homozygous GG with 65%, homozygous minor type CC with 6% and heterozygous minor type GC with 26% frequency distribution. High prevalence of GG genotype in the selected population presenting that GG genotype may have higher risk for susceptibility to HBV infection. Our results showed significant correlation of HMGB1 polymorphism with HBV infection in the selected Pakistani population.
Open Access: Yes
Bridging the gap between point-of-care and laboratory standards: comparative evaluation of MedSenso and DSA glucometers against Cobas analyzers for accurate diabetes monitoring
Publication Name: Brazilian Journal of Biology
Publication Date: 2025-01-01
Volume: 85
Issue: Unknown
Page Range: Unknown
Description:
Diabetes mellitus remains a major global health burden, with effective management relying heavily on accurate blood glucose monitoring. Personal glucometers are widely used for daily self-checks, yet their performance must be rigorously validated against laboratory standards to ensure reliability. This study undertook a diagnostic evaluation of three glucometers, DSA, MedSenso, and the laboratory-based Cobas systems (C503 and Pro analyzers), to assess their precision and clinical applicability. In a cross-sectional design, 150 clinical samples from diabetic patients were analyzed using the DSA glucometer and Cobas C503, while an additional 200 diabetic blood samples were tested to compare MedSenso with the Cobas Pro analyzer. Ethical approval was obtained, and diagnostic parameters including sensitivity, specificity, correlation, and difference percentages were evaluated against the respective Cobas gold-standard systems. Results revealed nuanced but clinically meaningful findings. For the DSA glucometer, correlation with Cobas C503 ranged from 87.9% to 100%, with differences varying between 0.0% and 32.3%. Although entries with perfect correlation (100%) and no difference (0.0%) indicated excellent agreement, instances of high correlation coupled with significant differences highlighted systematic biases, particularly consistent over- or underestimation by the DSA device. Such discrepancies underscore the need for device-specific awareness to avoid misinformed clinical decisions. In contrast, the MedSenso glucometer demonstrated excellent agreement with Cobas Pro, showing a correlation coefficient of 0.984 and near-identical glucose results across tested samples. Its ease of use and rapid reporting make MedSenso especially promising for clinical settings where fast decision-making is essential. Collectively, the study underscores the complexity of glucose measurement in diabetes care. While the DSA glucometer requires cautious interpretation due to systematic biases, MedSenso emerges as a trustworthy and practical alternative for both clinical and routine use. These findings highlight the importance of balancing correlation strength and difference analysis in device selection, reinforcing the need for continuous validation against laboratory standards to ensure accurate and dependable diabetes management.
Open Access: Yes
In vitro Multi-targeted Anti-cancer Effects of Bavachinin in Papillary Thyroid Carcinoma Cell Line: Dual Pathway Inhibition and Cytokine Downregulation
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2025-01-01
Volume: 24
Issue: 1
Page Range: Unknown
Description:
Background: Thyroid cancer is the most common endocrine malignancy, with aggressive subtypes frequently demonstrating resistance to conventional therapies. Bavachinin, a natural flavonoid derived from Psoralea corylifolia, has exhibited anti-cancer activity in various tumor models; however, its effects on thyroid cancer remain largely undefined. Objectives: The aim of this study is to evaluate the anti-cancer activity of bavachinin in the papillary thyroid carcinoma TPC-1 cell line and elucidate its underlying molecular mechanisms. Methods: TPC-1 cells were treated with bavachinin (5 - 20 μM) for 24 - 72 hours. Cell viability was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay); morphological changes were visualized by confocal microscopy. Migration and invasion were analyzed by wound-healing and Transwell assays, respectively. Cytokine secretion was measured using enzyme-linked immunosorbent assay (ELISA). Gene and protein expression levels of protein kinase B (AKT), mechanistic target of rapamycin (mTOR), extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Apoptosis was confirmed by assessing the B-cell lymphoma-2-associated X protein (BAX)/B-cell lymphoma-2 (BCL-2) ratio and cleaved caspase-3 activity. All experiments were performed in triplicate, and data are presented as mean ± standard deviation (SD). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by Tukey’s post-hoc test (P < 0.05). Results: Bavachinin significantly reduced cell viability, migration, and invasion in a dose-dependent manner (20 μM reduced viability by approximately 50% at 72 hours, P < 0.01). It suppressed the phosphorylation of AKT and ERK1/2, downregulated mTOR expression, and decreased secretion of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β). Apoptosis was confirmed by an increased BAX/BCL-2 ratio and elevated cleaved caspase-3 levels. Conclusions: Bavachinin exerts multi-targeted anti-cancer effects in thyroid carcinoma cells through dual inhibition of the phosphoinositide-3-kinase (PI3K)/AKT/mTOR and mitogen-activated protein kinase (MAPK)/ERK pathways, along with suppression of pro-inflammatory cytokines, culminating in apoptosis and impaired invasiveness.
Open Access: Yes
DOI: 10.5812/ijpr-166894
Innovative public-health strategies for neurodegenerative disease: leveraging diversified ultraviolet irradiation as a next-generation therapy
Publication Name: Brazilian Journal of Biology
Publication Date: 2025-01-01
Volume: 85
Issue: Unknown
Page Range: Unknown
Description:
Neurodegenerative diseases (NDs) such as Alzheimer’s, Parkinson’s, Huntington’s disease, and amyotrophic lateral sclerosis are escalating worldwide, straining healthcare systems and leaving patients with therapies that are largely palliative. Emerging evidence positions diversified ultraviolet (UV) irradiation as a groundbreaking, non-invasive strategy to counter these disorders. Beyond its traditional use in sterilization, specific UV spectra, UV-B (280–320 nm), UV-C (200–280 nm), and far-UV (207–222 nm), are now recognized for modulating oxidative stress, restoring mitochondrial function, correcting apoptotic dysregulation, and enhancing DNA repair. Innovative approaches such as riboflavin-mediated phototherapy and photobiomodulation (PBM) show the capacity to disaggregate toxic protein aggregates like β-amyloid and α-synuclein, boost antioxidant defenses, stimulate neurotrophic factors, and quell neuroinflammation. Preclinical models and early clinical trials reveal preserved cognition, enhanced neurogenesis, and reduced disease biomarkers, suggesting real translational promise. From a public-health perspective, UV-based interventions offer a cost-effective, scalable option for aging populations and resource-limited settings, especially when integrated with community-level health technologies and remote delivery platforms. Continued investigation of optimal dosing, long-term safety, and mechanistic pathways will be pivotal to unlock the full therapeutic and population-wide impact of this novel modality.
Open Access: Yes
Dual TYK2/JAK1 Inhibition by Brepocitinib Reprograms Synoviocyte Pathobiology: Mechanistic Insights Into Targeted Therapy for Rheumatoid Arthritis
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background: Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by synovial hyperplasia, persistent inflammation, and joint destruction. Targeted inhibition of intracellular signaling pathways, such as JAK-STAT, has improved RA treatment outcomes, though safety and selectivity remain as concerns. Brepocitinib, a dual TYK2/JAK1 inhibitor, has shown clinical efficacy in the management of autoimmune diseases, yet its mechanistic impact on synoviocytes remains underexplored. Objectives: To investigate the molecular and functional effects of brepocitinib on MH7A and RA-FLS synoviocytes, a key effector cell type in RA pathogenesis. Methods: MH7A and RA-FLS cells were treated with brepocitinib (0.5 µM, 1 µM, and 5 µM) for 24 hours. Cell viability was assessed. Western blotting was used to examine phosphorylation of TYK2, JAK1, STAT1/3, and apoptotic markers (BAX, BCL-2, caspase-3). Quantitative PCR and ELISA were performed to evaluate mRNA and protein levels, respectively, of IL-6, TNF-α, and IFN-γ. Wound healing assays measured synoviocyte migration. Results: Brepocitinib maintained ≥ 85% cell viability across all doses, compared with ~20% viability in doxorubicin-treated controls. At 5 µM, phosphorylation of JAK1 and STAT3 was suppressed by > 80%, while TYK2 and STAT1 inhibition reached ~70%. IL-6 and TNF-α transcripts were reduced by > 80% and IFN-γ by ~70%, with corresponding decreases in secreted cytokines (IL-6: 100 pg/mL to 20 pg/mL; TNF-α: 150 pg/mL to 15 pg/mL; IFN-γ: 41 pg/mL to 11 pg/mL). Brepocitinib shifted the BAX/BCL-2 ratio fourfold in favor of apoptosis and increased cleaved caspase-3 levels to ~80% of maximal response. Functionally, it reduced wound closure from ~75% in controls to ~20% at 5 µM, confirming potent inhibition of synoviocyte migration. Conclusions: Brepocitinib exerts multi-faceted effects on RA synoviocytes by simultaneously inhibiting inflammatory signaling, suppressing cytokine expression, restoring apoptotic sensitivity, and reducing migratory potential. These findings provide mechanistic support for brepocitinib as a targeted therapeutic agent in RA.
Open Access: Yes
DOI: 10.5812/ijpr-166019
MEK1/2 Inhibitor (U0126) and PI3K Inhibitor (LY294002) Suppress Herpes Simplex Virus Type 1 Replication by Targeting MAPK/ERK1/2 and PI3K/AKT Signaling Pathways: Implications for Oral Health and Translational Control of Orolabial HSV-1 Infection
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2025-01-01
Volume: 24
Issue: 1
Page Range: Unknown
Description:
Background: Current antivirals for orolabial Herpes simplex virus type 1 (HSV-1) often provide incomplete suppression and limited reactivation control, sustaining recurrent oral lesions and inflammation that compromise oral health. HSV-1 subverts host signaling networks to enhance its replication and trigger inflammation. Among these, the extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathways are hijacked to facilitate viral gene expression and cell survival. Objectives: In this study, we employed U0126 [a mitogen-activated protein kinase 1/2 (MEK1/2) inhibitor] and LY294002 [a phosphatidylinositol 3-kinase (PI3K) inhibitor] as targeted pharmacological tools to intercept HSV-1’s exploitation of host keratinocyte signaling. Methods: Human HaCaT keratinocytes were infected with HSV-1 and treated with U0126 or LY294002. Western blotting was used to assess phosphorylation of ERK1/2 and activation of protein kinase B (AKT). MTT assays were performed to evaluate cell viability. Real-time PCR was utilized to quantify viral transcripts (ICP0, ICP4, gB, and gC) and inflammatory cytokines [interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α)]. Confocal microscopy was employed to visualize the intracellular distribution of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), phosphorylated activation of protein kinase B (p-AKT), and HSV-1 glycoprotein D (gD). Viral titers were determined using plaque assays. Results: The HSV-1 infection induced a time-dependent increase in phosphorylation of ERK1/2 and AKT, with p-ERK1/2 peaking at 12 h and p-AKT increasing 2.5-fold by 24 h. Cell viability declined from 100% at baseline to 45% at 24-hours post-infection (hpi). Treatment with U0126 and LY294002 reduced p-ERK1/2 and p-AKT levels to 25% and 30% of infected controls, respectively, restoring viability to 82 - 86%. Both inhibitors markedly suppressed viral gene expression (ICP0, ICP4, gB, gC down by 60 - 80%) and inflammatory cytokines (IL-6 and TNF-α reduced by > 50%). Plaque assays showed a strong decline in infectious titers — from 175 plaques per well in untreated infection to 60 and 45 plaques after U0126 and LY294002, respectively. Confocal imaging further revealed diminished nuclear accumulation of p-ERK1/2 and p-AKT, indicating disruption of post-entry signaling critical for viral replication. Conclusions: Targeting host signaling bottlenecks with U0126 and LY294002 offers a dual-pronged antiviral strategy against HSV-1 by dismantling the ERK/AKT axis critical for replication and inflammatory amplification. These findings position MEK1/2 and PI3K as promising therapeutic nodes for managing cutaneous HSV-1 infections. This host-directed dual-pathway inhibition may therefore help reduce recurrent orolabial HSV-1 lesions.
Open Access: Yes
DOI: 10.5812/ijpr-164639
Curcumin modulates NF-κB and Nrf2 pathways to mitigate inflammation, oxidative stress, and virulence in Pseudomonas aeruginosa-infected bronchial epithelial cells
Publication Name: Archives of Microbiology
Publication Date: 2026-06-01
Volume: 208
Issue: 6
Page Range: Unknown
Description:
Pseudomonas aeruginosa pneumonia is characterized by excessive inflammation, oxidative damage, and virulence-driven epithelial injury. Targeting host inflammatory and antioxidant pathways, alongside bacterial virulence factors, may offer a complementary strategy to mitigate infection-induced damage. Curcumin, a bioactive compound from Curcuma longa, possesses established anti-inflammatory and antioxidant properties, but its effects on epithelial responses during bacterial infection remain incompletely defined. This study evaluated the impact of curcumin on NF-κB and Nrf2 signaling, inflammatory and oxidative responses, bacterial virulence, and epithelial repair in a cell-based model of P. aeruginosa–induced airway injury. Human bronchial epithelial BEAS-2B cells were pretreated with curcumin (10 µM) prior to infection with P. aeruginosa (1 × 10⁷ CFU/mL; MOI = 10). Cell viability, oxidative stress, cytokine production, bacterial load, and epithelial regeneration were assessed using MTT, DCFDA, qPCR, ELISA, while Western blotting and confocal microscopy characterized molecular and morphological changes. Curcumin pretreatment improved cell viability (~ 87% vs. ~64% in infected), attenuated NF-κB activation and cytokine expression, and enhanced Nrf2 pathway activity (HO-1, NQO1). Reactive oxygen species levels were reduced (~ 250% to ~ 140%), LasR expression and intracellular bacterial burden were decreased, and epithelial wound closure were partially restored compared with infected controls. Collectively, these findings indicate that curcumin modulates inflammatory, oxidative, and virulence-associated pathways and supports epithelial repair in P. aeruginosa-infected bronchial epithelial cells, suggesting potential relevance as a host-directed adjunct rather than a direct antimicrobial agent.
Open Access: Yes
Soluble factors from Aspergillus fumigatus promote NF-κB/AKT/ERK activation and pro-tumor phenotypes in lung cancer cells in vitro
Publication Name: Archives of Microbiology
Publication Date: 2026-06-01
Volume: 208
Issue: 6
Page Range: Unknown
Description:
Role of environmental fungi and Aspergillus fumigatus in respiratory diseases remains evident; however, its contribution to directly influencing lung cancer progression remains obscure. In this study, we investigated the effect of Aspergillus fumigatus extract (AFE) on the development of tumor-promoting characteristics in human lung cancer cell lines. The organism was distinguished based on Lactophenol Cotton Blue staining and further distinguished with protein expression patterns via SDS-PAGE and BCA analysis. A549 and H1299 lung adenocarcinoma human cell lines were challenged with AFE, and various cellular responses were monitored simultaneously for cell viability, proliferative activity, inflammatory gene expression, DNA damage expression, and migratory responses. AFE caused increased cell viability and exhibited cellular characteristics of highly proliferating cells with significant expression of Cyclin D1 and c-MYC. Highly inflammatory gene expression responses and protein expressions of AKT, ERK1/2, and NF-κB signaling pathways were noticed at both the gene and protein expression levels with NF-κB nuclear translocation verified with confocal microscopy studies. DNA damage expression markers like γ-H2AX, p-ATM, and p53 significantly contributed with observable genomic DNA cleavage. Additionally, AFE-exposed cells exhibited faster wound closure and expression of Epithelial-mesenchymal transition-associated factors contributing to cell migration and therapeutic efficacy of this combined approach needs further investigation and development into a targeting therapeutic agent against lung cancer.
Open Access: Yes
nc886 noncoding RNA regulates hepatitis B virus replication via PKR-dependent eIF2α phosphorylation
Publication Name: Frontiers in Cellular and Infection Microbiology
Publication Date: 2026-03-31
Volume: 16
Issue: Unknown
Page Range: Unknown
Description:
Objective(s) – Hepatitis B virus (HBV) replication is tightly controlled by host stress and innate immune pathways. The small noncoding RNA nc886 (vtRNA2-1) is a known endogenous inhibitor of protein kinase R (PKR), but its role in HBV biology remains unclear. This study aimed to define the function of the nc886–PKR–eIF2α axis in HBV-replicating hepatoma cells and to determine whether nc886 depletion suppresses HBV replication via PKR-dependent translational control. Materials and methods – Huh7 cells and Huh7 cells stably harboring a 1.3-mer HBV replicon were used. Endogenous nc886 and PKR expression was assessed by RT-qPCR and Western blot. Loss-of-function experiments employed two independent siRNAs against nc886 and one siRNA against PKR, alone or in combination, with scramble siRNA as control. PKR activation was induced by Poly(I:C); PKR and integrated stress response (ISR) were pharmacologically modulated using C16 (PKR inhibitor) and ISRIB (eIF2B activator), at non-toxic doses defined by MTT assay. Intracellular HBV DNA was measured by Southern blot, HBV pgRNA and subgenomic RNAs by Northern blot and RT-qPCR, and secreted HBsAg/HBeAg by ELISA. PKR–eIF2α–ATF4 signaling was evaluated by Western blot. Results – nc886 and PKR were efficiently and specifically knocked down without affecting cell viability. nc886 silencing in Huh7–HBV cells increased PKR-dependent eIF2α phosphorylation and ATF4, reduced HBV pgRNA and subgenomic RNAs, and decreased intracellular HBV DNA and secreted HBsAg/HBeAg. PKR knockdown alone slightly enhanced HBV readouts and completely rescued nc886-mediated inhibition of HBV replication and ISR activation in dual-knockdown cells. C16 or ISRIB restored HBV DNA, RNA and antigen production in nc886-silenced or Poly(I:C)-treated cells, while having no effect in control cells, indicating that rescue depended on ISR modulation. Conclusion – nc886 acts as a critical negative regulator of PKR-dependent ISR signaling during HBV replication in Huh7 cells. Its depletion activates PKR and eIF2α, imposing a translational block that suppresses HBV gene expression. The nc886–PKR–eIF2α module represents a novel host regulatory axis with potential relevance for host-directed HBV therapies.
Open Access: Yes
Wedelolactone Inhibits Hepatitis B Virus Replication by Modulating NF-κB and Nrf2/HO-1 Signaling: An in-vitro Huh7 1.3-mer HBV Plasmid Model
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background: Chronic hepatitis B virus (HBV) infection is a well-recognized cause of hepatic injury through prolonged viral replication, inflammation, and oxidative stress. Existing antiviral drugs limit viral replication but cannot eliminate viral transcription or even totally preclude liver injury, thus reemphasizing the significance of drugs with combined antiviral and hepatoprotective effects. Objectives: To evaluate the effects of wedelolactone on HBV replication, gene expression, inflammation, and oxidative stress in an in-vitro model of HBV plasmid transfection with human hepatic cells. Methods: Human hepatocellular carcinoma cells (Huh7) were transfected with a 1.3-mer plasmid and treated with wedelolactone (2.5 - 10 µM). Luciferase assays for HBV promoter activity, Northern blotting and Southern blotting for transcripts and replicative intermediates, qPCR for extracellular HBV DNA, and western blotting for viral antigens such as HBx were performed. Cell cytotoxicity was measured. NF-κB/IκB, inflammatory cytokines (TNF-α, IL-6), and antioxidant markers (Nrf2, HO-1, Keap1) were assessed to evaluate inflammatory and oxidative responses. Results: Wedelolactone significantly suppresses HBV promoter activity, RNAs, core particle formation, and extracellular HBV DNA. It reduced the expression of HBcAg and HBsAg. It inhibited NF-κB activation and cytokine release, while simultaneously enhancing Nrf2/HO-1 signaling, including induction of heme oxygenase-1 by lowering levels of Keap1. Conclusions: Wedelolactone exerts dual antiviral and hepatoprotective actions by inhibiting HBV replication and modulating inflammatory and oxidative stress pathways.
Open Access: Yes
DOI: 10.5812/ijpr-168329
Baicalein Protects H9c2 Cardiomyoblasts Against LPS-Induced Inflammatory Injury by Modulating the NF-κB/NLRP3 Inflammasome Pathway and Mitochondrial ROS
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background: Sepsis-related cardiomyopathy is mainly induced by uncontrolled inflammation and mitochondrial oxidative damage triggered by endotoxins, especially those of Gram-negative bacteria. Lipopolysaccharide (LPS), an endotoxin, activates the Toll-like receptor 4/nuclear factor kappa B (TLR4/NF-κB) signaling pathway and the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which further activates caspase-1 to process the pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18). Chronic activation of this signaling pathway is responsible for mitochondrial damage and loss of membrane potential. Baicalein, a flavonoid derived from the herbal plant Scutellaria baicalensis is well recognized for its anti-inflammatory and antioxidant properties, although its full modulation of the NF-κB/NLRP3 mitochondrial signaling pathway is not thoroughly understood. Objectives: In this study, we assessed whether baicalein has a protective effect on H9c2 cardiomyoblasts against inflammatory and mitochondrial injury induced by LPS through regulation of inflammatory pathways involving NF-κB, the inflammasome, caspase-1 activity, cytokine expression, and oxidation. Methods: H9c2 cells were pre-exposed to different baicalein concentrations (5 - 20 µM) and, after 1 hour, exposed to 1 µg/mL of LPS. Analysis included cell viability measurements by MTT assay, lactate dehydrogenase (LDH) release, and microscopic evaluation by phase-contrast microscopy. Expression levels of premature Nlrp3, Il1B, Il18, and Casp1 (0 - 4 h) were analyzed by real-time polymerase chain reaction (PCR). Protein levels of inflammasome and NF-κB pathway proteins were evaluated by immunoblot analysis. Activity of caspase-1 and cytokine secretion of IL-1β and IL-18 were evaluated by colorimetry and enzyme-linked immunosorbent assay (ELISA), respectively. Mitochondrial membrane potential (ΔΨm) was evaluated by staining with JC1 dye, and reactive oxygen species (ROS) levels were evaluated by fluorescence with DCFH2DA; N-acetylcysteine (NAC) was used as an antioxidant positive control. Results: LPS strongly elevated the transcription of Nlrp3 and Il1b at 2 hours, in addition to enhancing protein expression of NLRP3, activity of caspase-1, and secretion of IL-1β and IL-18. Underlying these pro-inflammatory responses were mitochondrial depolarization, augmented ROS production, diminished cell survival, and cytotoxicity. Prior administration of baicalein mitigated NF-κB activation, diminished priming of the inflammasome response at both transcriptional and protein levels, and decreased caspase-1 activity and secretion of the cytokines. Moreover, baicalein maintained mitochondrial membrane potential and diminished ROS within the cells, comparable to NAC, and improved cellular viability and morphology Conclusions: Baicalein confers substantial protection against LPS-induced inflammatory and oxidative damage in H9c2 cardiomyoblasts by concurrently suppressing NF-κB activation, mitigating NLRP3 inflammasome signaling, curbing caspase-1–driven cytokine maturation, and stabilizing mitochondrial function through reduction of ROS. These findings identify baicalein as a promising candidate for targeting the ROS-inflammasome axis in sepsis-associated cardiac dysfunction and related inflammatory cardiac disorders.
Open Access: Yes
DOI: 10.5812/ijpr-169689
Acylglycerol Kinase 2-mediated Inhibition of Sirtuin 2 Restores AMPK/AKT/mTOR Signaling Balance in Podocytes: A Pharmacological Strategy for Diabetic Nephropathy
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background: Diabetic nephropathy is a major cause of end-stage renal disease, driven in part by molecular dysfunctions in podocytes. Sirtuin 2 (Sirt2), a cytoplasmic NAD+-dependent deacetylase, has emerged as a potential regulator of key metabolic pathways, but its specific role in podocyte biology remains poorly defined. Objectives: This study aimed to investigate the function of Sirt2 in human podocytes (hPodo), delineate its interaction with histone deacetylase 6 (HDAC6), and evaluate the therapeutic potential of Sirt2 inhibition in restoring metabolic balance and protecting against diabetic nephropathy-associated podocyte stress. Methods: Comparative expression analysis was performed between hPodo and HEK293T kidney cells. Pharmacological inhibition of Sirt2 was carried out using acylglycerol kinase 2 (AGK2), alongside siRNA-mediated Sirt2 knockdown. AMPK/AKT/mTOR signaling activity was assessed by Western blotting and functional assays to determine metabolic and growth responses. Results: Human podocytes exhibited significantly elevated Sirt2 expression and high levels of HDAC6, forming a unique Sirt2–HDAC6 regulatory complex. Inhibition or silencing of Sirt2 induced robust AMPK activation while suppressing AKT/mTOR signaling. This signaling reprogramming restored energy sensing and attenuated hyperactive growth pathways, alleviating podocyte stress. Acylglycerol kinase 2 treatment reestablished metabolic homeostasis by disrupting Sirt2-mediated repression of AMPK. Conclusions: Sirtuin 2 inhibition, particularly through AGK2, emerges as a novel pharmacological strategy to protect podocytes, restore metabolic regulation, and potentially slow the progression of diabetic nephropathy. Significance Statement By inhibiting one of the important intracellular signaling pathways in human kidney cells, we could reduce the cellular stress that is commonly observed in diabetic kidney injury. This could serve as a drug target to slow the progression of kidney disease associated with diabetes mellitus.
Open Access: Yes
DOI: 10.5812/ijpr-165603
Next-generation bacteriophage therapeutic systems: CRISPR-based engineering, near-infrared bioimaging, and precision strategies for treating multidrug-resistant and extensively drug-resistant bacterial infections
Publication Name: Frontiers in Microbiology
Publication Date: 2026-01-01
Volume: 17
Issue: Unknown
Page Range: Unknown
Description:
The rapid rise of multidrug-resistant and extensively drug-resistant bacterial infections has renewed interest in bacteriophages as adaptable, targeted antimicrobials. Recent advances in phage engineering, including CRISPR-based approaches, now make it possible to refine host range, strengthen lytic performance, and deliver genetic payloads that target clinically important resistance determinants such as blaNDM, mecA, and mcr-1. In parallel, jumbo phages with large genomes often encode additional functions that support replication and biofilm disruption, offering practical advantages in densely structured infections where antibiotics perform poorly. A second limitation in phage translation has been measurement: in most settings, dosing and treatment duration remain guided by indirect endpoints rather than real-time information on distribution and activity. Near-infrared bioimaging addresses this gap by enabling noninvasive tracking of infection burden and phage kinetics in vivo through bacteriophytochrome-derived reporters, including iRFPs, miRFPs, and PAiRFPs. In this review, we bring these developments together and discuss how CRISPR-enabled phage engineering, jumbo-phage biology, and near-infrared readouts can be integrated into a precision framework that is measurable, adaptable, and clinically interpretable. We examine evidence across major drug-resistant pathogens, including Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, Burkholderia cepacia, and Mycobacterium abscessus. We also summarize practical constraints that remain central to clinical translation, manufacturing quality, host immune neutralization, and regulatory variability, and outline a realistic development pathway in which engineered phages and companion diagnostics progress from animal models to carefully defined clinical indications. Together, these advances support a shift from empirical phage use toward a more standardized, data-driven approach to treating drug-resistant infections.
Open Access: Yes
Metformin Attenuates Angiotensin II-Induced Cardiac Inflammaging-Like Injury Through Coordinated Nrf2 Activation and NF-κB Suppression
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background: Age-related cardiomyocyte vulnerability is driven by a convergent triad of persistent oxidative stress, chronic low-grade inflammation, and senescence-like signaling, which together accelerate functional decline and maladaptive remodeling. Angiotensin II (Ang II), a clinically relevant stress effector, amplifies reactive oxygen species (ROS) production and inflammatory activation, thereby impairing cardiomyocyte survival and repair capacity. Although metformin has emerging cardiovascular benefits beyond glycemic control, its coordinated capacity to counteract Ang II-driven inflammaging-like injury through the coupled regulation of antioxidant and anti-inflammatory pathways in human cardiomyocytes remains insufficiently defined. Objectives: This study aimed to determine whether metformin attenuates Ang II-induced injury and impaired repair in human AC16 cardiomyocytes and to evaluate whether this protection is associated with coordinated activation of Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2) and suppression of Nuclear Factor-κB (NF-κB). Methods: Human AC16 cardiomyocytes were preconditioned with metformin (0.5 or 1.0 mM) for 2 h and then challenged with Ang II (0.1 - 2.0 μM) under continuous metformin exposure. The primary efficacy endpoints were cell survival, quantified using the MTT assay, and repair competence, assessed by scratch wound closure. Secondary mechanistic endpoints included inflammatory mediators (TNFA, IL6, and IL1B); senescence-associated markers (CDKN2A/p16 and CDKN1A/p21); antioxidant genes linked to Nrf2 signaling (NFE2L2/Nrf2, HMOX1/HO-1, and NQO1); NF-κB pathway activation; antioxidant and apoptosis-related proteins; Nrf2 compartmentalization; and intracellular ROS. Results: Angiotensin II induced a dose-dependent injury phenotype in AC16 cardiomyocytes, reducing viability from 100.0 ± 1.1% in control cells to 64.9 ± 1.6% at 1.0 μM and 53.0 ± 1.6% at 2.0 μM, while markedly impairing wound closure (80.0 ± 2.0% in control vs 32.3 ± 2.5% with Ang II). Metformin attenuated this injury in a concentration-dependent manner, restoring viability to 81.7 ± 1.0% with 0.5 mM and 89.7 ± 1.0% with 1.0 mM and improving wound closure to 52.3 ± 2.5% and 70.0 ± 2.0%, respectively. Angiotensin II also robustly increased inflammatory cytokine expression, with TNFA, IL6, and IL1B increasing to 27.86 ± 2.79-fold, 29.86 ± 2.99-fold, and 29.86 ± 2.99-fold, respectively, accompanied by NF-κB activation, ROS accumulation (187.7 ± 2.5% of control), apoptosis-associated signaling, and upregulation of the senescence-like markers p16 and p21. Metformin markedly suppressed these responses, reducing cytokine expression toward near-baseline levels, lowering ROS to 120.0 ± 2.0%, decreasing the BAX/BCL-2 ratio from 2.98 to 1.19, and reducing cleaved caspase-3 and cleaved PARP to 120% and 112% of control, respectively. In parallel, metformin enhanced Nrf2-associated antioxidant signaling, increasing NFE2L2 to 2.14 ± 0.17-fold, HMOX1 to 4.29 ± 0.30-fold, and NQO1 to 3.81 ± 0.27-fold, consistent with enhanced antioxidant defense under Ang II stress. Conclusions: Metformin attenuated Ang II-driven cardiac inflammaging-like injury and impaired repair, in association with reduced inflammatory signaling and enhanced activation of antioxidant pathways. These findings support a mechanistically coherent model in which NF-κB suppression and Nrf2 activation may contribute to metformin-mediated protection; however, direct pathway dependence requires confirmation through perturbation-based studies.
Open Access: Yes
DOI: 10.5812/ijpr-170947
SIRT1 and IFI16 cooperatively regulate HBV replication via epigenetic modulation and innate immune activation
Publication Name: Future Virology
Publication Date: 2026-01-01
Volume: 21
Issue: 6
Page Range: 393-404
Description:
Background: Hepatitis B virus (HBV) establishes persistent infection through the formation of a covalently closed circular DNA (cccDNA) minichromosome, which enables immune evasion and sustained viral replication. While SIRT1, IFI16, and STING are known to modulate antiviral pathways, their cooperative role in regulating HBV transcription and host immunity remains poorly defined. Methods: We investigated the functional interplay between SIRT1, IFI16, and STING using confocal microscopy, chromatin immunoprecipitation (ChIP), gene silencing, and quantitative PCR in HBV-replicating cell models. Interactions with HBV cccDNA and effects on viral transcription and interferon-stimulated gene (ISG) expression were analyzed. Northern blotting and RT-qPCR were used to evaluate HBV RNA output, including total HBV RNA and 3.5 kb RNA, in order to provide both transcript-pattern and quantitative assessment of transcriptional changes. Results: SIRT1 physically interacted with IFI16, with increased colocalization during HBV replication. Both proteins were recruited to HBV cccDNA, promoting transcriptionally active chromatin. Silencing SIRT1 or IFI16 disrupted cccDNA binding and reduced viral RNA levels. Although STING was not cccDNA-associated, its antiviral activity via ISG induction was dependent on IFI16 knockdown produced the strongest reduction in both total HBV RNA and 3.5 kb RNA, supporting a cooperative role for these factors in sustaining HBV transcriptional output. Although STING was not cccDNA-associated, its antiviral activity via ISG induction was dependent on IFI16. Triple knockdown of SIRT1, IFI16, and STING showed the strongest suppression of ISG expression. Conclusion: SIRT1 and IFI16 enhance HBV gene expression through epigenetic regulation of cccDNA, while also supporting STING-mediated antiviral responses. These findings reveal a dual regulatory axis governing HBV persistence and identify potential targets for antiviral therapy.
Open Access: Yes
Andrographolide Alleviates Mycoplasma pneumoniae Pneumonia in Children by Inhibiting Alveolar Epithelial Cell Pyroptosis Through the HMGB1/TLR4/NF-κB Pathway
Publication Name: Iranian Journal of Pharmaceutical Research
Publication Date: 2026-12-01
Volume: 25
Issue: 1
Page Range: Unknown
Description:
Background:Mycoplasma pneumoniae pneumonia (MPP) is a pulmonary inflammatory disease caused by Mycoplasma pneumoniae (Mp) infection that primarily involves the bronchi, alveoli, and pulmonary interstitium. It is the most common cause of community-acquired pneumonia in children. Andrographolide (AG) is a natural diterpenoid lactone with anti-inflammatory and immunomodulatory activities; however, its specific role and mechanism in MPP remain unclear. Objectives: This study aimed to investigate the mechanism by which AG inhibits pyroptosis in alveolar epithelial cells via the HMGB1/TLR4/NF-κB signaling pathway in the treatment of MPP in children. Methods: Mp-stimulated MLE-12 cells were treated with AG at 10, 20, or 50 μM. Cell viability and injury were assessed using CCK-8 and LDH assays, respectively. Protein expression levels of NLRP3, HMGB1, TLR4, and p-NF-κB p65 were determined by Western blotting, and cytokine levels (IL-6 and TNF-α) were measured by ELISA. In an Mp-infected mouse model, mice received AG at 25 or 50 mg/kg. Body weight, lung index, lung histopathology, pathway-related protein expression, and cytokine levels (IL-1β and TNF-α) in bronchoalveolar lavage fluid (BALF) were evaluated. Results: Mp significantly induced cytotoxicity, pyroptosis, and inflammation in vitro and in vivo (all P < 0.001). Andrographolide treatment dose-dependently reduced LDH release, suppressed HMGB1/TLR4/NF-κB/NLRP3 activation, and decreased proinflammatory cytokine levels (P < 0.05). In mice, AG improved survival-related metrics, ameliorated lung pathology, and inhibited pathway activity and cytokine secretion. Conclusions: Andrographolide mitigates MPP via the HMGB1/TLR4/NF-κB signaling pathway by inhibiting pulmonary epithelial cell pyroptosis and inflammation. These findings indicate its potential as a therapeutic agent.
Open Access: Yes
DOI: 10.5812/ijpr-169826
