Research in Pharmaceutical Sciences最新文献

Background and purpose: Non-alcoholic fatty liver disease (NAFLD) is a chronic metabolic condition characterized by the accumulation of excess fat in the liver, which can ultimately lead to fibrosis and cirrhosis. This study investigated the impact of resveratrol on the signaling pathways miR-141/SIRT1/AMPK/TGF- p/Smad3 in fatty liver of male C57/BL6 mice.

Experimental approach: Twenty-one male C57/BL6 mice were acclimatized for 10 days and divided into 3 groups (n = 7), including control, NAFLD, and NAFLD + resveratrol groups. After an 8-week HFD to induce NAFLD, the mice were treated with resveratrol (100 mg/kg/day; oral gavage) for 8 weeks. At the end of the study (16 weeks), serum and liver tissue samples were collected. Gene expression was assessed using RT- PCR, while protein levels were analyzed via Western blotting. Statistical analysis was performed using SPSS 16.

Findings/results: The results of the study showed that the expression levels of the genes Smad3 and miRNA- 141 were significantly reduced in the resveratrol-treated group compared to the NAFLD group, while the expression levels of SIRT1 and TGF-β were significantly increased. In addition, the Western blot results indicated that the levels of the proteins P-AMPK and SIRT1 in the resveratrol-treated group were significantly higher compared to the NAFLD group. Furthermore, a significant reduction in fat accumulation and degeneration was observed in the histopathological findings of the liver in the resveratrol-treated group.

Conclusion and implications: The study concluded that resveratrol has the potential to reduce liver damage from NAFLD by modulating various signaling pathways, particularly TGF-β/Smad3, SIRT1/AMPK, and miRNA-141, leading to improved lipid metabolism and reduced hepatic steatosis. While the findings underscored the multifaceted therapeutic effects of resveratrol, further research and clinical trials are necessary to fully understand its mechanisms and applications in humans.

Background and purpose: One of the most prevalent types of malignancies affecting the cells in the mucosal surface of the oral cavity and pharynx regions is head and neck squamous cell carcinoma (HNSCC). This study analyzed the metabolic profile of genes involved in the metabolism of fatty acids (FAs) to identify biomarkers with prognostic and diagnostic potential in HNSCC.

Experimental approach: Gene set enrichment analysis, differential gene expression, and correlation analysis methods were used to examine the enrichment and expression patterns of genes involved in the metabolism of FAs in the HNSCC tissue samples. Gene ontology and network analysis were performed to explore the molecular interactions in the metabolic pathways of FAs. The diagnostic and prognostic potentials of identified highly dysregulated genes in HNSCC were examined by ROC test and Cox-regression methods.

Findings/results: FA-associated metabolic pathways were significantly dysregulated in the HNSC cancer samples. For the diagnosis of HNSC cancer, CYP4B1 and FMO2 could be potential biomarkers, while for the prognosis of HNSCC survival periods, ACOX2, CYP4F12, and ELOVL6 could hold valuable biomarker potential.

Conclusion and implications: The findings could help target the metabolism of FAs using the identified biomarkers for the design of new therapeutic opportunities for patients with HNSCC.

Background and purpose: Curcuma aeruginosa rhizome essential oil (CREO) is widely used in traditional medicine owing to its diverse biological activities. However, no information regarding its potential toxicity is available. This study aimed to evaluate the potential acute and sub-acute oral toxicities of CREO in Sprague Dawley rats.

Experimental approach: CREO was isolated via steam distillation and characterized using GC/MS. For acute toxicity, rats were divided into four groups and administered CREO at 2, 4, 8, and 16 g/kg. For the sub-acute evaluation, 30 male and 30 female rats were divided into 6 groups (1 control, 3 treatment doses, and 2 satellite), with doses of 50, 100, and 200 mg/kg BW administered for 28 days.

Findings/results: GC/MS analysis identified eucalyptol, camphor, and epicurzerenone as the main phytochemically active components in CREO. The acute toxicity test demonstrated that CREO was toxic only at very high doses, with a lethal dose (LD₅₀) of 5662 mg/kg of body weight. Evaluation of sub-acute toxicity showed no significant changes in body weight, hematological, biochemical, and histopathological parameters in rats receiving CREO at doses < 200 mg/kg. However, rats that received CREO at 200 mg/kg showed liver early abnormalities. Similar to most natural extracts, CREO showed a hormetic dose response.

Conclusion and implications: This study suggests that CREO can be safely administered orally for therapeutic purposes at controlled doses. However, prolonged consumption and/or high doses may pose potential risks. Further evaluations are required to determine possible long-term effects.

Background and purpose: Single-chain variable fragments (scFvs) offer advantages over full-length monoclonal antibodies in cancer therapy, including reduced size, lower production costs, and easier handling. However, Escherichia coli (E. coli) often leads to the formation and aggregation of inclusion bodies (IBs). This study aimed to optimize the expression and purification of an anti-CD22 scFv (CD22-scFv) in E. coli and evaluate its functional properties.

Experimental approach: The CD22-scFv construct was subcloned into pET-28a(+) and expressed in E. coli strains Rosetta (DE3) and Rosetta-gami 2. To overcome IBs formation, two purification methods were employed to enhance soluble protein production: hybrid conditions, a novel one-step immobilized metal affinity chromatography (IMAC)-based on-column refolding method was employed, using gradually decreasing urea and increasing imidazole concentrations; native conditions, expression parameters (IPTG concentration, post-induction temperature, and time) were optimized, followed by IMAC. The CD22-scFv binding to CD22 antigen and its anti-proliferative effects on target cells were assessed via flow cytometry and MTT assay.

Findings/results: CD22-scFv was successfully expressed in Rosetta (DE3) but not Rosetta-gami 2. Hybrid purification yielded 15.86 mg/L protein, outperforming native purification (3.65 mg/L). Flow cytometry confirmed the binding of native- and hybrid-purified CD22-scFv to CD22 Raji cells with 75.5% and 55.8% efficiency, respectively. Native-purified CD22-scFv significantly inhibited Raji cell proliferation while sparing CD22^- cells.

Conclusion and implications: This study established a scalable and cost-effective strategy for producing functional CD22-scFv with high specificity and anti-proliferative effects. The findings highlight its potential for targeted therapies and diagnostics, warranting further in vivo and clinical studies.

Background and purpose: The Klotho (Klo) gene, an aging suppressor in rats, accelerates aging when disrupted and extends lifespan when overexpressed. It encodes a transmembrane protein primarily expressed in renal tubules. This study investigated the protective effects of central Klo, both alone and in combination with cholinergic anti-inflammatory pathway (CAP) inhibition, against ischemia-reperfusion injury (IRI)- induced acute kidney injury. The current study evaluated the expression of inflammatory and anti-inflammatory genes (including Illb, Tnfa, Tgfb, Trem2, and Il10) in the kidney, alongside plasma levels of creatinine (Cr), blood urea nitrogen (BUN), and signs of acute tubular injury.

Experimental approach: Klo was microinjected into the rostral ventrolateral medulla, and CAP inhibition was achieved through intraperitoneal administration of mecamylamine (Mec). Real-time RT-PCR and hematoxylin and eosin staining were used for gene expression analysis and histopathological examination, respectively.

Findings/results: The results showed elevated Cr and BUN levels, tubular injury, and increased inflammatory gene expression in IRI and IRI + Mec groups, as well as reduced Il10 in the IRI + Mec group. Klo exhibited protective effects. Elevated Tgfb expression was seen in IRI + Klo and IRI + Mec + Klo groups one week post-surgery.

Conclusion and implications: These findings indicated Klo potential to extend lifespan and protect against age-related diseases, including kidney disease and inflammation, via neural modulation of peripheral immunity.

Background and purpose: Colorectal cancer (CRC) is the second leading cause of cancer death. While surgery and medicines offer complete treatment, recurrence and medication resistance pose challenges. This study assessed the cytotoxic impact of entinostat, a histone deacetylase (HDAC) inhibitor, and genistein, a soybean isoflavone, combination on CRC cells.

Experimental approach: The cytotoxic effect of genistein, combined with entinostat, was tested in HCT-116 and HT-29 cell lines, along with their impact on migration and colony formation. Gene expression of the cell cycle regulatory protein CDC25A was assessed using qPCR.

Findings/results: The IC50 values of genistein, entinostat, and their combination in HCT-116 cells were 24.48 μM, 13.65 μM, and 14.55 μM, respectively. In HT-29, the IC50 values were 30.41 μM, 20.25 μM, and 19.98 μM, respectively. In the HT-29 cell line, a 1:1 ratio of entinostat and genistein resulted in a combination index of 0.6 using a concentration of 1.56 μM of each compound, indicating a synergistic effect. In contrast, no synergistic effect was produced between the two drugs in the HCT-116 cell line. In HCT-116 cells, genistein, entinostat, and their combination significantly reduced wound closure compared to the control. In contrast, in HT-29 cells, only the combination treatment was effective, while genistein and entinostat alone showed no notable impact. In HCT-116, entinostat, genistein, and their combination reduced the number of colonies significantly compared to the control, while in HT-29, only entinostat and the combination reduced the number of colonies significantly compared to the control. Furthermore, the combination of genistein with entinostat was more effective in reducing CDC25A expression in the HT-29 cells compared to entinostat treatment alone.

Conclusions and implications: Combining genistein with entinostat could potentiate the entinostat cytotoxic effect in CRC.

Background and purpose: The Fat mass and obesity-associated protein (FTO) plays a significant role in esophageal cancer by regulating N6-methyladenosine (m6A) modification. FTO inhibition has shown potential in cancer therapies but remains underexplored. This study aimed to identify a safer, FDA-approved compound for FTO inhibition that can be used in combination with chemotherapy drugs.

Experimental approach: FDA-approved drugs were screened from the Zinc 15 database using AutoDock Vina against the 3D structure of FTO (PDB ID: 3LFM). Discovery Studio software was used to determine binding interactions. The GROMACS package was used for molecular dynamics simulations. A non-toxic concentration was determined through an MTT assay on KYSE-30 esophageal cancer cells. The ELISA assay was used to measure the m6A levels in RNA.

Findings/results: Four compounds, ergotamine, midazolam, digoxin, and loratadine, were identified. Loratadine (ΔG: -8.9) formed stable interactions with FTO, specifically with residues Ser229, Tyr109, Leu109, Val229, and His231. Molecular dynamic simulations of the FTO-loratadine complex revealed higher RMSD fluctuations (0.4-0.6 nm), but the system remained stable overall. RMSF analysis showed similar fluctuation patterns in all three systems, indicating that loratadine did not affect protein structure stability. MM/PBSA calculations revealed powerful binding energy for the FTO-loratadine complex (-135.73 kJ/mol), driven by favorable van der Waals interactions. KYSE-30 cells treated with loratadine (100 μM), m6A levels in KYSE- 30 cells compared to the control group were significantly elevated at a non-toxic concentration.

Conclusion and implications: Loratadine is a promising, low-toxic FTO inhibitor that could complement chemotherapy for esophageal cancer.

Background and purpose: Nanotechnology can improve drug delivery by enhancing cell selectivity, releasing at specific target sites, and improving bioavailability while reducing adverse events and potential treatment costs. The current study aimed to synthesize curcumin/magnesium oxide (Cur/MgO) nanoparticles (NPs) and evaluate their neuroprotective effects in a mouse model of ketamine-induced neurotoxicity.

Experimental approach: XRD, FE-SEM, and a particle size analyzer determined the average crystalline and particle sizes. UV-Vis examined absorption patterns, and FT-IR spectroscopy analyzed the functional groups involved in the reaction. To evaluate the effectiveness of Cur/MgO NPs on ketamine-induced neurotoxicity, male BALB/c mice were divided into 7 groups and received the following treatments (intraperitoneally, daily for 2 weeks). Groups 1 and 2 received normal saline (0.2 mL) and ketamine (25 mg/kg). Group 3 received curcumin (40 mg/kg) and ketamine (25 mg/kg). Groups 4-6 received ketamine (25 mg/kg) and Cur/MgO NPs (10, 20, and 40 mg/kg). Group 7 received MgO (5 mg/kg) and ketamine (25 mg/kg). Finally, the hippocampal tissues were examined morphologically and analyzed for oxidative stress, inflammation, apoptotic markers, and mitochondrial quadruple complex enzymes.

Results/findings: Both Cur/MgO NPs and curcumin reduced IL-1β, TNF-α, Bax, and MDA levels and GSSG content and increased GSH, Bcl-2, GPx, GR, and SOD. Cur/MgO NPs and curcumin also increased mitochondrial quadruple complex enzymes and inhibited histological changes in the dentate gyrus and CA1 hippocampus areas in ketamine-induced neurotoxicity.

Conclusion and implications: Cur/MgO NPs were more neuroprotective against the ketamine-induced histomorphological changes, inflammation, apoptosis, and oxidative stress than curcumin alone.

Background and purpose: This study aimed to evaluate the cytotoxicity of synthesized thiosemicarbazone derivatives, particularly on biomarkers associated with ferroptosis.

Experimental approach: Thiosemicarbazone derivatives (C1-C5) were synthesized by condensation between thiosemicarbazide and the corresponding benzaldehyde derivatives. The compounds were characterized using IR spectroscopy and H/C NMR spectroscopy. To evaluate their biological activity, PC-12 cells were cultured in DMEM/MEM medium supplemented with 10% bovine serum albumin. Cell viability was assessed using the MTT assay, while intracellular reactive oxygen species (ROS) levels were measured using DCFH-DA. Additionally, glutathione peroxidase (GPX) activity, lipid peroxidation (LPO), and total antioxidant capacity (TAC) were evaluated to determine oxidative stress and antioxidant response.

Findings/results: In cell viability assessments, C2 exhibited the highest toxicity, while C4 demonstrated the lowest toxicity after 24 h. Among all derivatives, only C3 reduced ROS levels without affecting GPX activity. All derivatives effectively reduced LPO, although C5 showed the least effectiveness in this regard. In contrast to C2 and C5, TAC was significantly higher than the control after treatment with C1, C3, and C4.

Conclusion and implications: These findings suggest that thiosemicarbazone derivatives may influence the ferroptosis cell death pathway through their chelation properties, necessitating further research on their ability to bind to iron. Their effects on oxidative stress and cellular antioxidant capacity provide valuable insights for therapeutic strategies.

Background and purpose: Rheumatoid arthritis (RA) is a chronic inflammatory disease associated with systemic complications and progressive disability. Systemic side effects and poor drug delivery to joints limit current treatments. This study aimed to enhance the efficacy of tofacitinib (Tofa) by synthesizing novel pH-triggered biocompatible polymers, both folate-targeted and non-folate-targeted.

Experimental approach: First-generation polymers were synthesized and characterized using FT-IR and ¹HNMR spectroscopy. The critical micelle concentration of the copolymers was evaluated, and Tofa-loaded micelles were prepared using the dialysis method. The physical properties of the micelles were assessed using FE-SEM and dynamic light scattering. Cytotoxicity of Tofa/chondroitin sulfate-maleic-dexamethasone (Tofa/CHS-Mal-DEX) and Tofa/folic acid-polyethylene glycol-chondroitin sulfate-maleic-dexamethasone (Tofa/FA-PEG-CHS-Mal-DEX) micelles was evaluated on the fibroblastic L929 and RAW264.7. The cellular uptake and anti-inflammatory effects were investigated in the activated Raw 264.7 cell line.

Findings/results: Tofa/CHS-Mal-DEX and Tofa/FA-PEG-CHS-Mal-DEX micelles exhibited particle sizes of 188 nm and 173.06 nm, respectively, with entrapment efficiencies of 51% and 72.76%. The release profiles exhibited that about 40% of Tofa was released from micelles over 62 h in physiological pH, whereas in acidic conditions, this significantly decreased to 2 h. Micelles demonstrated improved uptake efficiency, resulting in a significant reduction in IL-6 levels compared to free Tofa. None of the micelle formulations indicated cytotoxic effects on fibroblastic L929 and Raw 264.7 macrophage cell lines.

Conclusion and implications: The developed folate and non-folate-targeted micelles were not toxic and biocompatible for enhancing the therapeutic potential of Tofa in RA and improving drug delivery.