Akkas, Onder; Celebi, Demet; Celebi, Ozgur; Baser, Sumeyye; Kalayci, Sıla Ektas; Ari, Betül; Guler, Mustafa Can; Dabanlıoglu, Bulent; Celebi, Selahattin; Kılıclıoglu, Metin

Gut microbiota plays a crucial role in maintaining host homeostasis by regulating metabolic processes and immune responses. Disruptions in microbial composition are closely associated with inflammatory diseases and are often linked to the activation of key signaling pathways such as Toll-like receptor 4/myeloid differentiation primary response 88/nuclear factor kappa TLR4/MyD88/NF-κB and NLR family pyrin domain-containing 3 (NLRP3) inflammasome. Natural bioactive compounds, particularly flavonoids, have gained attention due to their potential to modulate both gut microbiota and inflammation-related pathways. In this context, the present study aimed to evaluate the effects of gossypetin on gut microbiota composition and its regulatory role on TLR4, MyD88, NF-κB, and NLRP3 signaling pathways in a rat model. Adult female Wistar albino rats were divided into control and gossypetin-treated groups (50 mg/kg, oral gavage/56 days dose). Gut microbiota was analyzed by 16S rRNA sequencing, and protein expression levels were assessed using Western blot. Histopathological, immunohistochemical, and immunofluorescence analyses were also in liver, intestinal, and spleen tissue performed. Gossypetin administration reduced microbial diversity and altered microbiota composition, with increases in Mediterraneibacter spp., Blautia spp., and Lactobacillus spp. Western blot results showed significant decreases in NLRP3 (p ≤ 0.01) and NF-κB (p ≤ 0.05) levels, while TLR4 and MyD88 remained unchanged. Histological analyses revealed mild tissue alterations and increased oxidative stress markers. These results suggest that gossypetin modulates microbiota composition and exerts selective anti-inflammatory effects, highlighting its potential in microbiota-associated inflammatory regulation.

ATASEVER, Aslıhan; ÇELEBİ, Fikret; YILDIRIM, Serkan; BOLAT, İsmail; ÇINAR, Burak

In this study, the potential protective effects of syringic acid (SA) on gastric tissue were investigated in an indomethacin (INDO)-induced gastric ulcer model. A total of 84 male Sprague–Dawley rats were randomly divided into seven groups. In the in vivo experiments, rats were administered SA at doses of 5, 50, and 100 mg/kg and omeprazole (OMP) at a dose of 5 mg/kg intragastrically (i.g.) for 14 days, and indomethacin (100 mg/kg, i.g.) was administered on the final day. Following INDO administration, the rats were sacrificed under anesthesia, and gastric tissues were carefully excised for further analyses. The collected gastric tissues were subjected to biochemical, histopathological, and immunofluorescence analyses. In addition, in silico analyses were performed to support the INDO-induced gastric ulcer model. Using the licensed Schrödinger Maestro 2025/1 software, the binding properties of INDO to the COX-1 receptor were evaluated through molecular docking, MM-GBSA, and pharmacophore matching analyses. INDO administration was associated with oxidative stress, inflammation, apoptosis, and histopathological damage in gastric tissue. SA treatment appeared to alleviate INDO-induced gastric injury through its antioxidant, anti-inflammatory, and anti-apoptotic properties. SA treatment ameliorated histopathological alterations in ulcerated areas, particularly at doses of 50 and 100 mg/kg, whereas the 5 mg/kg dose did not show a significant protective effect. In addition, in silico analyses suggested that INDO may contribute to ulcer formation by inhibiting COX-1, thereby reducing prostaglandin production in the gastric mucosa. Overall, the findings of this study suggest that SA may reduce oxidative stress, suppress inflammatory responses, and inhibit apoptosis, thereby contributing to the protection of gastric tissue against INDO-induced injury. These results indicate that SA may have therapeutic potential for the prevention of NSAID-induced gastric injury; however, further experimental and clinical studies are needed to confirm these effects and clarify the underlying mechanisms.

Bal Taştan, Tuğba; Selli, Jale; Toktay, Erdem; Üstündağ, Hilal; Ugan, Rüstem Anıl

Polycystic ovary syndrome (PCOS) adversely affects endometrial receptivity and fertility outcomes. This study investigated whether combining high-dose antioxidants (astaxanthin or omega-3 fatty acids) with metformin improves endometrial receptivity markers, reduces oxidative stress, and normalizes hormone profiles in a PCOS rat model compared to metformin alone or no treatment. Fifty-six female Sprague–Dawley rats (200-250g) were divided into seven groups: Control, PCOS, PCOS + Met, and groups receiving metformin plus either astaxanthin (PCOS + Met + AX5 or PCOS + Met + AX10) or omega-3 (PCOS + Met + OMG400 or PCOS + Met + OMG1000). PCOS was induced using letrozole for 21 days, followed by one week of treatment. After establishing endometrial receptivity, uterine tissues were analyzed histopathologically, immunohistochemically, and biochemically on day 5. The PCOS group demonstrated significant increases in body weight and oxidative stress markers (malondialdehyde) compared to controls (p < 0.001). High-dose antioxidant supplementation (PCOS + Met + AX10 and PCOS + Met + OMG1000) significantly improved these parameters (p < 0.001). Immunohistochemically, integrin β3 expression increased while mucin 1 (MUC-1) decreased in high-dose groups compared to PCOS (p < 0.001). The LH:FSH ratio normalized to control levels in PCOS + Met + AX10 and PCOS + Met + OMG1000 groups. Histological examination revealed enhanced endometrial morphology and increased pinopode-like structure formation in antioxidant-supplemented groups. High-dose astaxanthin and omega-3 fatty acid supplementation, combined with metformin, may enhance endometrial receptivity in PCOS by reducing oxidative stress and modulating key implantation markers. This therapeutic approach could potentially improve fertility outcomes in women with PCOS.

Doganay, Songul; Üstündağ, Hilal; Erbaş, Elif; Yanar, Sevinc; Hayme, Serhat; Kurt, Nezahat

Sepsis-induced brain dysfunction remains a significant challenge with limited therapeutic options, warranting exploration of multi-target approaches. Methylene blue (MET) is a phenothiazine derivative with mitochondrial-enhancing and electron transport chain regulatory properties, while nicotinamide adenine dinucleotide (NAD) serves as a crucial cofactor in cellular energy metabolism and redox reactions. This study investigated the therapeutic effects of MET and NAD, individually and in combination, in lipopolysaccharide (LPS)-induced sepsis model. Male Sprague–Dawley rats (n = 35) were randomized into five groups: control, LPS (8 mg/kg), LPS + NAD (250 mg/kg), LPS + MET (15 mg/kg), and LPS + NAD + MET. Brain tissues were analyzed for oxidative stress, inflammation, and blood–brain barrier integrity. The combination therapy demonstrated the most pronounced improvements across clinical parameters, antioxidant capacity evidenced by significantly elevated superoxide dismutase levels (p < 0.001) and cerebral histopathological outcomes. It also significantly reduced myeloperoxidase activity (p < 0.001) compared to the LPS group. Pro-inflammatory cytokines IL-6 and IL-8 were significantly decreased in both MET and NAD monotherapy groups (p < 0.05), while TNF-α showed a decreasing trend without statistical significance. Brain-derived neurotrophic factor levels were maintained in the combination group, comparable to controls, while TGF-β showed differential modulation across treatment groups. Immunohistochemical analysis revealed LPS-induced blood–brain barrier disruption, evidenced by reduced occludin expression (p < 0.01), increased aquaporin-4 expression in perivascular astrocytic end-feet (p < 0.05), and elevated glial fibrillary acidic protein reactivity in protoplasmic astrocytes (p < 0.01). In conclusion, MET and NAD individually provided significant protection against LPS-induced brain injury by reducing neuroinflammation, oxidative stress, and neuronal damage. The combined treatment offered incremental benefits in certain outcomes. These findings suggest that both agents may have therapeutic potential in SAE.

Danışman-Kalındemirtaş, Ferdane; Sert, Esra; Kariper, İ. Afşin; Korkut, Işık Neslişah; Erdem-Kuruca, Serap

Although chemotherapeutic agents such as doxorubicin (DOX) are used to treat leukemia, their application is often limited by systemic toxicity, low specificity, and the development of drug resistance. The development of drug delivery systems can overcome the problems associated with conventional delivery methods. In this study, innovative albumin-based UV-DOX-NPs were synthesized by UV irradiation and evaluated for their leukemia inhibitory activity; they represent a novel formulation strategy that has seen limited application to date. Their effects on leukemic Jurkat and THP-1 cells were extensively studied. Advanced techniques such as dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDX) were used for characterization. UV-DOX-NPs exhibited increased cytotoxicity and significantly lower IC50 values compared to free DOX. Apoptosis rates were also higher in both cell lines following DOX-NP treatment. Flow cytometry revealed a significant downregulation of leukemia-associated surface markers, including CD5, CD7, and CD3 in Jurkat cells and CD7, CD14, and CD33 in THP-1 cells. In addition, UV-DOX-NPs significantly reduced the expression of ABCG2, an important MDR-associated transporter. Our results demonstrate that UV-synthesized, albumin-based UV-DOX-NPs not only enhance antileukemic efficacy but also modulate the leukemic immunophenotype and overcome multidrug resistance. These results provide a promising platform for a more targeted and safer leukemia therapy.

Ereminsoy, Ergün; Demir, Yeliz; Yıldırım, Sümeyra Tuna; Türkeş, Cüneyt; Küfrevioğlu, Ömer İrfan

In the present study, novel Schiff base ligands were synthesized via the condensation of salicylaldehyde derivatives with o-aminophenol, and their metal complexes with Co(II), Ni(II), Cu(II), Zn(II), Cd(II), and UO₂(VI) metal actetates were successfully prepared. The compounds were comprehensively characterized using FT-IR, 1H-NMR, 13C-NMR, UV–Vis spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and magnetic susceptibility measurements. FT-IR spectra confirmed azomethine (C = N) coordination through characteristic bands at 1612–1630 cm−1, while 1H-NMR spectra exhibited diagnostic azomethine proton signals at δ 8.84–8.62 ppm. XRD analysis revealed a semi-crystalline nature with prominent reflections at 2θ = 18.02° and 20.87°, and SEM images demonstrated nanorod-like morphologies. Magnetic moment values supported the proposed coordination geometries of the paramagnetic complexes. The biological activity of the synthesized compounds was investigated through in vitro inhibition studies against lipoxygenase (LOX), a ~ 97 kDa enzyme purified from Chenopodium quinoa Willd. with 77.89% purity and 7.24% yield. All compounds exhibited competitive inhibition toward LOX, with inhibition constants (Ki) in the range of 0.014 ± 0.002 to 0.858 ± 0.194 μM, indicating remarkable inhibitory potency. Molecular docking studies further substantiated the experimental results by revealing key hydrogen-bonding and π-cation interactions between the most active compounds and essential amino acid residues within the LOX active site. Overall, the results demonstrate that these Schiff base metal complexes represent promising candidates for the development of potent LOX inhibitors.

Halici, Hamza; Karakoy, Zeynep; Yurdgulu, Emir Enis; Ay, Yusuf Anil; Cadirci, Elif; Toktay, Erdem; Kavas, Betul

Ovarian torsion is a gynecological emergency caused by rotation of the vascular pedicle, leading to ischemia, necrosis, and potential fertility loss. Management remains controversial, particularly regarding oophorectomy versus ovarian preservation. Detorsed ovaries continue cytokine secretion, impairing contralateral ovarian function, and no protective therapy currently exists. This study aimed to compare oophorectomy and detorsion strategies and to determine whether post-detorsion long-term pirfenidone therapy preserves ovarian function, thereby providing experimental resolution to the clinically pivotal “save or sacrifice” dilemma. Forty female rats were divided into five groups: Healthy, IR, IR + OVX, IR + Pirfenidone 50 mg/kg, and IR + Pirfenidone 100 mg/kg. Ischemia was induced in the left ovary of IR groups; in IR + OVX, the ovary was excised at 3 h, while in other groups, ovaries were reperfused. Treatments continued for 30 days. Ovarian tissues and blood were analyzed histopathologically, biochemically, and molecularly. Ovarian ischemia–reperfusion induced a persistent inflammatory and profibrotic response not only in the ischemic ovary but also in the contralateral ovary, leading to reduced estrogen levels and marked histopathological damage. Importantly, detorsion alone did not prevent contralateral injury, whereas oophorectomy attenuated molecular alterations. Post-detorsion pirfenidone therapy dose-dependently restored estrogen production, suppressed inflammasome activation (IL-1β, NLRP3, caspase-1) and fibrotic signaling (SMAD2, SMAD3, TGF-β1), and preserved normal follicular architecture in both ovaries. These findings demonstrate that the detorsed ovary remains a biologically active source of injury and identify pirfenidone as a potential adjunct therapy capable of protecting ovarian function after torsion.

Gul, Merve; Alagoz, Mehmet Abdullah; Hepokur, Ceylan; Algul, Oztekin

To design, synthesize, and biologically evaluate a novel series of 1-(4-chlorophenyl)-2-(4-methylpyrazol-1-yl)ethenone oxime derivatives (5–15) as targeted anticancer agents, focusing on their selectivity for glioma, mechanistic action, and blood–brain barrier (BBB) permeability. The oxime derivatives were synthesized and characterized using 1H-NMR, 13C-NMR, and mass spectrometry. In vitro cytotoxicity was assessed against C6 (glioma), MCF-7 (breast cancer), and SH-SY5Y (human neuroblastoma) cell lines. Flow cytometry evaluated apoptosis and cell cycle arrest for the lead compound. Molecular docking simulations were performed against CDK6 (PDB: 5L2I) and P-glycoprotein/ABCB1 (PDB: 7A6E). All derivatives exhibited selective potency against C6 glioma cells. Compound 14 demonstrated the highest antiproliferative activity (IC₅₀: 31.23 µM) against tumor lines while sparing SH-SY5Y cells. Flow cytometry confirmed that compound 14 triggers apoptosis and induces cell cycle arrest in C6 cells. Molecular docking predicted the most favorable docking scores for compound 14 against both CDK6 and P-glycoprotein. Structure–activity relationships (SAR) analysis validates the 4-methylpyrazole core as a viable scaffold for targeted glioma therapy. These results suggest that further functionalization of the O-alkyl oxime moiety merits further investigation as a starting point for potentially brain-penetrant scaffolds, although in vivo and target-level validation are required.

Manavoğlu Kirman, Esra; Bolat, İsmail; Karaarslan, Tuba; Tekin, Samet; Atasever, Aslıhan

Lead acetate (PbAc) is an environmental toxicant known to induce liver injury through oxidative stress, inflammation, and apoptosis. This study aimed to investigate the potential hepatoprotective effects of β-caryophyllene (BCP), a natural sesquiterpene with antioxidant and anti-inflammatory properties, against PbAc-induced hepatotoxicity and to explore the underlying molecular mechanisms. Sixty male Sprague–Dawley rats were randomly assigned to five groups: control, BCP200, PbAc, PbAc + BCP100, and PbAc + BCP200. PbAc was used to induce hepatotoxicity, while BCP was administered at doses of 100 and 200 mg/kg. Serum liver enzymes (ALT, AST, ALP, and GGT), oxidative stress parameters (MDA, SOD, and GSH), and inflammatory cytokines (TNF-α, IL-1β, and IL-6) were evaluated. Immunohistochemical and immunofluorescence analyses were performed to assess Nrf2, HO-1, Sirt1, Keap1, and 8-OHdG expression. In addition, the mRNA expression of PI3K, AKT, mTOR, Bax, Bcl-2, and Caspase-3 was determined by qRT-PCR, and histopathological examinations of liver tissue were conducted. PbAc exposure significantly increased serum liver enzyme levels, lipid peroxidation, and pro-inflammatory cytokines while reducing antioxidant defense markers. It also suppressed Nrf2/HO-1 signaling and PI3K/AKT/mTOR gene expression, accompanied by increased Bax and Caspase-3 and decreased Bcl-2 expression, indicating enhanced apoptosis. Histopathological analysis confirmed severe hepatic degeneration and necrosis. BCP treatment significantly attenuated these alterations by reducing oxidative stress and inflammatory responses, restoring Nrf2/HO-1 signaling, improving PI3K/AKT/mTOR pathway activity, and regulating apoptosis-related gene expression. These findings suggest that β-caryophyllene may exert protective effects against PbAc-induced hepatotoxicity through modulation of oxidative stress, inflammation, cell survival signaling, and apoptosis pathways, highlighting its potential as a supportive therapeutic candidate for heavy metal-induced liver injury.

Atila Uslu, Gözde; Uslu, Hamit; Çoban, Taha Abdulkadir; Özkaraca, Mustafa; Mendil, Ali Sefa; Aygörmez, Serpil

In the originally published version of this article, inaccuracies were identified in Figure 9. Specifically, the images in Figures 9C and 9D were incorrectly presented due to the misplacement of the representative image for Figure 9D. This error has now been corrected, and Figure 9D has been revised accordingly. Additionally, errors were found in the annotation of Figure 9. Figure 9B was mistakenly labeled as the Nobiletin group; however, it actually represents the negative control group (NFκB). Conversely, Figure 9C was incorrectly labeled as the negative control group, whereas it should be identified as the Nobiletin group (IL-1β). The legend of Figure 9 has been corrected to accurately reflect these revisions. These corrections do not affect the results or conclusions of the study. The corrected figure and legend are provided below. (Figure presented.) Fig. 9 A Negative control group IL-1β, B Negative control group NFκB, C Nobiletin group IL-1β, D Nobiletin group NFκB. (IL-1β and NFκB immune negativities.) IHC