Ahmet Cumaoğlu

Objective: This study examined the efficacy of hyperbaric oxygen therapy (HBOT) in facilitating recovery from high fructose-induced pulmonary fibrosis in rats and explored the underlying mechanisms. Methods: Male Wistar rats were randomly divided into four groups: Control (standard diet for 10 weeks), High fructose diet (HFD, for 10 weeks), Hyperbaric oxygen therapy (HBOT standard diet with 2.4 ATA (atmospheric pressure absolute) for 1 hr. during weeks 9–10) and HFD+HBOT (High fructose diet with 2.4 ATA for one h during weeks 9–10). Histopathological alterations, including mononuclear cell infiltration and hyperemia were determined by Hematoxylin and Eosin staining. Fibrosis was evaluated using Masson's trichrome staining. Markers of autophagy (LC3B: Microtubule-associated protein light chain 3B, Beclin-1) and inflammation (COX-1: Cyclooxygenase-1) were examined using immunohistochemical staining. Results: In rats, a high-fructose diet caused interstitial fibrosis, hyperemia, and mononuclear cell infiltration in the lung tissues. Additionally, HFD boosted the production of the proinflammatory COX-1 protein and autophagic LC3B and Beclin-1. The outcomes were comparable to those of the control group in rats administered hyperbaric oxygen therapy alone. Mononuclear cell infiltration, hyperemia, and fibrosis, along with autophagic and inflammatory markers, were markedly attenuated in the HFD+HBOT group. Conclusion: The key finding of our study is that HBOT effectively mitigates the development of pulmonary fibrosis in rats fed a HFD. These findings underscore the potential of HBOT as a promising therapeutic intervention for managing pulmonary fibrosis associated with metabolic disorders.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors have gained attention for their potential therapeutic applications in cancer treatment. Recent Mendelian randomization and observational analyses support the protective role of SGLT2 inhibition in reducing prostate cancer risk. Additionally, SGLT2 expression in prostate cancer patient samples has been confirmed through immunohistochemistry. The therapeutic potential of empagliflozin, a selective SGLT2 inhibitor, in treating prostate cancer, either alone or with chemotherapeutic agents like docetaxel, remains largely unexplored. This study investigated the cytotoxic and synergistic effects of empagliflozin in combination with docetaxel in LNCaP and DU- 145 prostate cancer cells. Cell viability was assessed using the MTT assay, and synergy was evaluated using the Chou-Talalay method. Western blot analysis was conducted to examine the effects of empagliflozin, alone and in combination with docetaxel, on key molecular targets, including p-AMPKα, p-p70S6 K1, p-PRAS40, and p-Akt. Empagliflozin exhibited concentration-dependent cytotoxic effects in both LNCaP and DU- 145 cells, with a higher potency observed in DU- 145 cells. When combined with docetaxel, empagliflozin demonstrated synergistic activity, as indicated by combination index values < 1. Empagliflozin upregulated p-AMPKα and downregulated p-p70S6 K1 and p-PRAS40. The combination with docetaxel further enhanced these effects. Notably, empagliflozin alone downregulated p-Akt in LNCaP cells but not in DU- 145 cells, highlighting cell-line-specific differences. Empagliflozin reduces prostate cancer cell viability and enhances the cytotoxic effects of docetaxel, suggesting a promising combination strategy for prostate cancer therapy. Additional in vivo studies and clinical trials are needed to assess the translational relevance of these findings.

Accumulating evidence suggests that sodium–glucose cotransporter 2 (SGLT2) inhibitors may be effective at eliminating tumor cells. While empagliflozin exhibits nearly the highest selectivity for SGLT2 over SGLT1, its specific impact alone and in combination with tamoxifen remains largely unexplored in estrogen receptor α-positive (ERα +) breast cancer. This study investigated the anticancer effects of empagliflozin and its potential synergy with tamoxifen in MCF-7 breast cancer cells. The individual and combined cytotoxic effects of empagliflozin and tamoxifen were assessed using the xCELLigence system. The activities of AMP-activated protein kinase α (AMPKα), p38 mitogen-activated protein kinase (p38 MAPKα), p70-S6 kinase 1 (p70S6K1), and protein kinase B (Akt) were assessed using Western blotting. The gene expression levels of peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) and Forkhead box O3a (FOXO3a) were assessed via qPCR. Our results revealed time- and concentration-dependent cytotoxic effects of empagliflozin and tamoxifen whether administered separately or in combination. While tamoxifen exhibits potency with an IC₅₀ value of 17 μM, approximately ten times greater than that of empagliflozin (IC₅₀ = 177 μM), synergistic effects are observed when the concentrations of the two agents approach their respective IC₅₀ values. Additionally, empagliflozin significantly increases AMPKα activity while concurrently inhibiting Akt, p70S6K1, and p38 MAPKα, and these effects are significantly enhanced when empagliflozin is combined with tamoxifen. Moreover, empagliflozin modulates the gene expression, downregulating PGC-1α while upregulating FOXO3a. Empagliflozin exerts anti-proliferative and anti-survival effects by inhibiting mTOR, Akt, and PGC-1α, and it exhibits synergy with tamoxifen in MCF-7 breast cancer cells. Graphical Abstract Proposed anticancer mechanism of empagliflozin in MCF-7 breast cancer cells.

In this study, we investigated the anti-microbial and anti-inflammatory activities of the cosmopolite macrolichens Usnea articulata (L.) Hoffm., Umbilicaria crustulosa (Ach.) Lamy and Bryoria fuscescens (Gyeln.) Brodo & D.Hawksw hydroalcoholic extracts to contribute the potential pharmacological uses of lichens. In vitro antimicrobial activities of ethanol extracts against Gram-negative bacteria Escherichia coli, Gram-positive bacteria Staphylococcus aureus, and the yeast Candida albicans were presented using the Broth microdilution method. The most effective lichen extract against gram-positive bacteria S. aureus was U. articulata ethanol extract with a MIC value of 0.125 mg/ml. U. articulata and B. fuscences extracts have similar anti-fungal activities despite having MIC values of 0.5 mg/ml. The antiinflammatory effects of the extracts on Lipopolysaccharide/Interferon-gamma (LPS/IF-γ) induced macrophage-like cellular systems (BV-2 microglia and RAW 264,7 macrophages) were evaluated by measuring P38 mitogen-activated protein kinase phosphorylation (P38MAPK), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase 2 (NOS2) mRNA and protein expression. Especially, Usnea and Umbilicaria extracts also attenuated the LPS/IF-γ induced increase in P38MAPK phosphorylation, COX-2, and NOS2 expression in both macrophage-like cells without any cytotoxicity. According to the results of our study, we suggest that the anti-inflammatory mechanism of lichen extracts might result from the inhibition of P38MAPK phosphorylation through a reduction in COX-2 and NOS2 expressions.

Psoralidin, a prenylated coumestan, has been reported its anti-inflammatory effect, but the regulatory effects on inflammasome activation and pyroptosis-related cytokines are not clear. The aim of this study was to investigate the regulatory role of psoralidin on inflammasome activation and expression of pyroptosis-related cytokines in Lipopolysaccharide (LPS)/Nigericin-stimulated RAW 264.7 macrophages. Regulatory mechanism of psoralidin on inflammasome activation was evaluated using Western blotting for NLRP3, adaptor protein apoptosis-associated speckle-like protein (ASC)and Caspase-1. Pyroptosis-related cytokines were evaluated using RT-PCR for interleukin-1β (IL-1β)and interleukin-18 (IL18).Non-cytotoxic concentration of Psoralidin (5.0µM) significantly inhibited LPS/Nigericin induced inflamasome activation. In addition, pretreatment with psoralidin suppressed the LPS/Nigericin-induced production of IL-1β and IL18. Our results indicate that the regulatory effects of psoralidin on inflammasome activation and pyroptosis related cytokine production in RAW 264.7 macrophages are associated with suppression of NLRP3inflammasome activation and inhibition of the release of pyroptosis related cytokines.According to these results, Psoralidine may be considered as a potential therapeutic candidate for the prevention of inflammatory diseases or to support the treatment of inflammation.

To evaluate hyperbaric oxygen therapy (HBO) based on ovarian histology, total antioxidant status (TAS), total oxidant status (TOS), and anti-müllerian hormone (AMH), in the ovarian insufiency (POI) model created with cyclophosphamide (CYP). The rats were separated into 3 groups of the control group (n=6), the CYP group (n=6), and the CYP+HBO group (n=6). The rats in the CYP group and the CYP+HBO group were injected intraperitoneally with 200 mg/kg CYP on day 1, followed by 8 mg/kg/day for 14 days to create POI. From the 15^th day onwards, the rats in the CYP+HBO group were placed in a hyperbaric cabin and exposed to 100% oxygen at 2.4 atm pressure for one h, and were then returned to their cages at the end of the hour. A statistically significant decrease was determined in the primordial and primary follicle counts in the CYP group compared with the control group (p<0.05). In the CYP+HBO group, a statistically significant increase was determined in the primordial and primary follicle counts (p<0.05). The serum AMH levels were seen to be significantly decreased in the CYP group compared with both the control group and the CYP+HBO groups. The HBO was seen to decrease TOS and increase TAS. HBO could be an alternative treatment to minimize the effect of ovarian follicle loss caused by CYP, which is used for treating tumors that commonly occur in young females of reproductive age.