Objective: As an intricate process, wound healing involves contributions from numerous cell types. Fibroblasts critically support wound healing via sustaining the integrity of the extracellular matrix, promoting collagen synthesis, and driving contraction during proliferation and remodeling. A balanced inflammatory response is essential for this repair. The apelinergic system, comprising Apelin and APJ (apelin receptor)—abundantly found in the skin and scar fibroblasts—plays a significant regulatory role. This is among the first to reveal the role of exogenous Apelin-13 in wound healing using a scratch model in CCD-1072Sk fibroblast cell line. Methods: The doses of 2 and 5-µg/ml Apelin-13 were chosen based on the cell viability assay result. After seeding cells in 24-well plates, the wound scratch model was applied once they reached approximately 90% confluency. Samples of the control (CTL), A-2, and A-5 groups were collected and analyzed at 0 h (hour) (baseline), 24 h, 48 h, and 72 h. After the wound scratch model, the wound gap area was checked at the relevant time points. ELISA analysis was performed for levels of TGF-β1, TNF-α, and IL-10. Apoptosis was analyzed with a fluorescent microscope using the Annexin V-FITC/PI method. Results: In the viability assay, 2 and 5 µg/ml Apelin-13 doses were used. Wound closure decreased significantly at all time points in both Apelin groups vs. 0 h (p<0.001). Apoptosis increased significantly in A-2-48 and A-5-48 groups (p<0.001). TNF-α decreased in A-2 vs. CTL (p<0.001) and A-5-72 (p<0.001). TGF-β1 increased in A-5-72 vs. A-2-72 (p<0.05). IL-10 increased in all Apelin groups vs. CTL (p<0.001). Conclusion: Apelin-13 promoted wound healing by balancing inflammation, enhancing fibroblast proliferation, migration, and reducing apoptosis. 2 µg/ml Apelin-13 showed notable effects, while 5 µg/ml was superior at certain time points. Apelin-13 appears to be a promising wound-healing agent.

Glutamate-mediated excitotoxicity leads to mitochondrial dysfunction, apoptosis, and neuronal cell death. This study aims to investigate whether NPY2 receptors (NPY2R) and NPY5 receptors (NPY5R) enhance the effects of Apelin-13/APJ signaling pathways as modulatory cofactors in the neuroprotection provided by Apelin-13 against excitotoxic damage and in the prevention of learning-memory disorders. D-Glutamic acid-induced excitotoxicity was established in 42 male Sprague–Dawley rats (6–8 weeks, 200-250 g). Animals were randomly divided into six groups (n = 7); Control (C; 0.9% NaCl, i.p), D-Glutamic Acid (G; 4 mg/kg, i.p), Apelin-13 (A; 300 µg/kg, i.p), D-Glutamic Acid + Apelin-13 (GA), D-Glutamic Acid + Apelin-13 + NPY2R antagonist (GAN2; 1,5 mg/kg, i.p) and D-Glutamic Acid + Apelin-13 + NPY5R antagonist (GAN5; 1,5 mg/kg, i.p). Locomotor activity were evaluated with the Open Field (OFT), short/long-term memory and learning performance, allocentric-egocentric orientation were assesed with novel object recognition (NORT) and Morris water maze (MWM) tests. All parameters were normalized to the C group, and statistical significance between groups was assesed. In group G, a significant decrease (p < 0.001) in Extracellular Signal Regulatory Kinase (ERK1/2) and Protein Kinase B-1 (AKT-1) levels and an increase (p < 0.001) in Caspase-3 were observed. Oxidative parameters increased in the G and GAN2 groups. Antioxidant parameters were also elevated in GA and GAN5, similar to C and A groups. An increase in MWM latency to the target quadrant (p < 0.001) and a decrease in NORT discrimination index (p < 0.001) were found in the G and GAN2 groups compare to the C and A. Histochemical staining scores showed that the protection of Apelin-13 was mediated by NPY2R. In GAN2, blocking NPY2R reduces Apelin-13’s neuroprotection, which is sustained only via NPY5R with limited effect. In GAN5, Apelin-13’s protection was enhanced through NPY2R, as shown with NPY5R blockade. Accordingly, Apelin-13 exerts its neuroprotective effects primarily through NPY2R, its modulatory influence via NPY5R appears to be comparatively limited. Graphical Abstract Graphical abstract illustrating the interplay between Apelin-13, NPY2 receptor (NPY2R) antagonist and NPY5 receptor (NPY5R) antagonist in modulating D-Glutamic acid induced excitotoxicity. Left side: This represents the normal glutamate excitotoxicity condition. Excessive glutamate release stimulates NMDA and AMPA receptors found in the postsynaptic membrane, leading to Na ⁺ and Ca ⁺² overload, ROS/RNS production, mitochondrial dysfunction, apoptosis, neuronal death, and cognitive impairment. Middle: Exogenous administration of Apelin-13 along with an NPY2R antagonist results in selective blockade of NPY2R while leaving NPY5R active. NPY2R blockade fails to inhibit presynaptic glutamate release, allowing continued activation of NMDA and AMPA receptors. Therefore, the neuroprotective effects of Apelin-13 are confined to NPY5R-mediated pathways. Right: When Apelin-13 is administered together with the NPY5R antagonist results in selective blockade of NPY5R while leaving NPY2R active. NPY2R activation inhibits presynaptic glutamate release, Apelin-13 also inhibits NMDA and AMPA receptors. Consequently, the neuroprotective effects of Apelin-13 are predominantly dependent on NPY2R. VGCC: Voltage-gated calcium channel; Glu: Glutamate; APJ: Apelin receptor; NPY2R/NPY5R: Neuropeptide Y receptors 2 and 5; NPY: Neuropeptide Y; ROS/RNS: Reactive oxygen/nitrogen species, AMPA: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; NMDA: N-methyl-D-aspartate; Ca ⁺² : Calcium, Na ⁺ : Sodium.

Background and Objectives: Type 1 Diabetes Mellitus (T1DM) is an autoimmune disease with T cell-mediated pathogenesis of pancreatic β-cell destruction, leading to insulin deficiency. MicroRNAs such as miR-223 and miR-106b, along with PTEN, have been reported to participate in the pathophysiology of diabetes and its complications. The current study has explored the expression of miR-223, miR-106b, and PTEN and their association with various clinical and biochemical parameters in subjects diagnosed with T1DM. Materials and Methods: Sixty T1DM patients (two groups as uncomplicated/ with microalbuminuria) and fifty healthy volunteers, age- and sex-matched, were enrolled in this study. The fasting venous blood samples were collected, and PTEN and miRNAs (miR-223 and miR-106b) levels were measured by ELISA and real-time PCR, respectively. Results: The PTEN levels of patients with microalbuminuria were significantly lower than those of patients without microalbuminuria, while those of miR-223 and miR-106b were significantly increased in the T1DM group compared with the healthy control group (p < 0.001). ROC analysis indicated that PTEN, miR-223, and miR-106b could be potential biomarkers for diagnosing T1DM with high specificity but with variable sensitivities. Also, PTEN and miR-223 were negatively correlated with r =−0.398 and p < 0.0001, indicating that they were interrelated in their role within the T1DM pathophysiology. Conclusions: In the current study, it has been shown that the circulating levels of PTEN, miR-223, and miR-106b are significantly changed in T1DM patients and may back their potential to be used as non-invasive biomarkers for the diagnosis and monitoring of T1DM. Low PTEN protein expression was related to high miR-223 expression, indicating involvement of these miRNA in the regulation of PTEN. Further studies should be performed to clarify the exact mechanisms and possible clinical applications of these molecules.

Metformin has been suggested to have protective effects on the central nervous system with an unclarified mechanism. The similarity between the effects of metformin and the inhibition of glycogen synthase kinase (GSK)-3β suggests that metformin may inhibit GSK-3β. In addition, zinc is an important element that inhibits GSK-3β by phosphorylation. In this study, it was investigated whether the effects of metformin on neuroprotection and neuronal survival were mediated by zinc-dependent inhibition of GSK-3β in rats with glutamate-induced neurotoxicity. Forty adult male rats were divided into 5 groups; Control, Glutamate, Metformin + Glutamate, Zinc Deficiency + Glutamate, Zinc Deficiency + Metformin + Glutamate. Zinc deficiency was induced with a zinc-poor pellet. Metformin was given orally for 35 days. D-glutamic acid was administered intraperitonally on the 35th day. On the 38th day, neurodegeneration was examined histopathologically and effects on neuronal protection and survival were evaluated via intracellular S-100β immunohistochemical staining. The findings were examined in relation to non-phosphorylated (active) GSK-3β levels and oxidative stress parameters observed in brain tissue and blood. Neurodegeneration was higher (p < 0.05) in rats fed with an inadequate zinc diet. Active GSK-3β levels were increased in groups with neurodegeneration (p < 0.01). Decreased neurodegeneration and increased neuronal survival (p < 0.01); decreased active GSK-3β (p < 0.01) levels and oxidative stress parameters, and increased antioxidant parameters were observed in groups treated with metformin (p < 0.01). Metformin had less protective effects in rats fed with an inadequate zinc diet. Metformin may demonstrate its neuroprotective effects and may increase S-100β mediated neuronal survival by a zinc-dependent inhibition of GSK-3β in glutamate neurotoxicity.

Objective: Cancer is a pathophysiological condition that develops as a result of DNA damage caused by environmental mutagens and is becoming increasingly prevalent. Genetic factors, the immune system, and cytokines are reported important task the pathophysiological mechanisms of breast cancer, mostly cancer among women today. Natural plants such as Colchicum speciosum (C. speciosum) are gaining prominence in alternative therapy research due to their colchicine content and anticancer properties. Our study was conducted to evaluate the immunomodulatory and anticancer effects of C. speciosum bulb extract on MCF-7 cell. Methods: C. speciosum was harvested and extracted; then the dose ratio was determined using the MTT method, and cytotoxicity, apoptotic level, and antibacterial activity were evaluated in MCF-7 cell. Cytokine levels were measured using commercially available kits via the ELISA method, and apoptotic assessment was examined using a commercial kit containing Annexin V-FITC/PI via fluorescence microscopy. The data was analysed statistically using GraphPad Prism (v10.2.0; GraphPad Software, ABD). Results: In the MTT evaluation, a significant decrease in cell proliferation was observed in the 240 and 500 µg/mL applications compared to the control group, with increasing dose rate and incubation time. The cytokine profile of C. speciosum bulb was characterized by decrease of pro-inflammatory cytokines (TNF-α, IL-1β) and increase in anti-inflammatory cytokines (IL-10), while a decrease in TGF-β1 and BDNF levels was detected depending on the applied dose. Annexin V-FITC/PI analyses demonstrated that C. speciosum bulb extract elicited dose- and time-dependent apoptotic responses in MCF-7 cell. In addition, antibacterial effects of C. speciosum bulb extract were observed. Conclusions The C. speciosum bulb was observed to have regulatory effects on cell proliferation via cytokines and apoptotic mechanisms in a dose- and time-dependent manner. Additionally, its antibacterial effect was observed independently of our findings.