Administration Of Nicotinamide Research Articles

A zebrafish model of nicotinamide adenine dinucleotide (NAD+) deficiency-derived congenital disorders.

Congenital NAD deficiency disorder (CNDD) is a multisystem condition in which cardiac, renal, vertebral, and limb anomalies are most common, but anomalies in all organ systems have been identified. Patients with this condition have biallelic pathogenic variants involving genes in the nicotinamide adenine dinucleotide (NAD+) synthesis pathway leading to decreased systemic NAD+ levels. CNDD anomalies mimic the clinical features described in vertebral-anal-cardiac-tracheoesophageal fistula-renal-limb (VACTERL) association raising the possibility that CNDD and VACTERL association possess similar underlying causes. However, the mechanism by which NAD+ deficiency causes CNDD developmental anomalies has not been determined, nor has NAD+ deficiency been definitively linked to VACTERL association. Therefore, additional animal models amenable to detailed observation of embryonic development are needed to address the causes and progression of congenital anomalies in both CNDD and VACTERL association. Here, we describe a zebrafish model of NAD+ disruption to begin to model CNDD and VACTERL association phenotypes, assessing developmental anomalies in real-time. Treatment of zebrafish embryos with 2-amino-1,3,4-thiadiazole (ATDA), a teratogen known to disrupt NAD+ metabolism, resulted in neural tube, craniofacial, cardiac, and tail defects. These defects were rescued by the administration of nicotinamide (NAM) in a dose-dependent manner. Our work establishes zebrafish as a useful model for investigating the mechanistic causes and developmental dynamics of CNDD and VACTERL association. Further, as VACTERL association has been linked to teratogens, our zebrafish model provides a platform to assess these agents.

The effect of a new phytocomposition based on the cranberry leaf extract and amino acids on the pancreas and liver of rats in the model of diabetes mellitus type 2

In pharmacotherapy of diabetes mellitus type 2 and metabolic syndrome, medicinal plants and amino acids are used as an important preventive and therapeutic supplement to correct disorders that develop against the background of insulin resistance, which determines the relevance of developing new domestic combined antidiabetic herbal medicines. Aim. To study the effect of a phytocomposition based on the polyphenol extract from large-fruited cranberry leaves and amino acids (L-arginine, taurine, glycine) on the state of the liver and pancreas of rats in an experimental model of diabetes mellitus type 2. Materials and methods. Diabetes mellitus type 2 was reproduced by a single injection of rats with a streptozotocin solution in the dose of 65 mg/kg, intraperitoneally with the previous (15 min) intraperitoneal administration of nicotinamide in the dose of 230 mg/kg against the background of obesity (keeping rats on a high-calorie diet for 12 weeks). The pancreatic and hepatoprotective effect of the phytocomposition was studied according to the morphofunctional state of pancreatic β-cells, the content of glycogen and neutral fats in the liver and morphometric indices in the pancreas. Results. The phytocomposition studied had a stimulating effect on regenerative processes in the insulin-producing apparatus of the rat pancreas – the severity of proliferation of small β-cells in pancreatic islets significantly increased compared to the control pathology. The phytocomposition at the morphological level showed hepatoprotective properties considerably reducing the manifestations of the toxic-dystrophic action of diabetogens, significantly restoring fat and carbohydrate metabolism in hepatocytes. Conclusions. The results obtained substantiate the feasibility of further study of the phytocomposition as a promising antidiabetic agent.

Role of Nicotinamide in the Pathogenesis of Actinic Keratosis: Implications for NAD+/SIRT1 Pathway.

Actinic keratosis (AK) is a precursor to invasive squamous cell carcinoma, making early diagnosis and treatment essential to prevent progression. Among available therapeutic options, nicotinamide (NAM) has shown potential in reducing AK progression. NAM is a precursor of nicotinamide adenine dinucleotide (NAD+), which activates sirtuin (SIRT)1, a protein with anti-cancer properties. Although the role of SIRT1 in AK is still debated, no data currently exist on the systemic modulation of this protein in AK. Therefore, this study aims to evaluate whether NAM, by increasing serum NAD+ levels, may promote SIRT1 activation in peripheral blood mononuclear cells (PBMCs) in AK patients. Thirty patients were enrolled and treated with NAM for 24 months. Hematological, biochemical, and skin condition assessments were conducted, alongside the measurement of SIRT1 and NAD+ levels. A decrease in basophils, monocytes, total cholesterol, and blood glucose levels was observed in the study group, along with a reduction in AK lesions. Notably, NAM treatment significantly enhanced serum NAD+ levels, and nuclear SIRT1 activity in PBMCs. In conclusion, NAM administration significantly reduced AK progression in a NAD+/SIRT1-dependent manner, supporting its role as a chemopreventive agent in AK management.

Hesperidin & Apigenin Attenuates Diabetes & Lipid Levels in Experimentally Induced Diabetes Mellitus in Wistar Rats.

The aim of the present investigation is to study the antidiabetic potential of Hesperidin and Apigenin in Diabetic rats. Wistar Albino rats of either sex (150 to 200 g) were purchased from the CPCSEA approved vendor New Delhi. A total of three animals were used, which received a single oral dose in 500 mg/kg, body weight of different flavonoids. Doses equivalent to 25 mg and 50 mg per kilogram body weight were calculated, and suspended in 1% w/v Tween 80 solutions for the experiment. For OGTT, Different doses of both flavonoids i.e. Hesperidin & Apigenin were administered 60 min prior to oral glucose load (2.0 g/kg). Diabetes was induced in overnight fasted rats by Streptozotocin (60 mg/kg), 15 min. after the i.p. administration of Nicotinamide (120mg/kg). Streptozotocin was dissolved in citrate buffer (0.1 M, pH 4.5) & nicotinamide was dissolved in normal saline. During the study period of 21 days, animals was weighed at 0, 7, 14 and 21 day and effect of vehicle, standard drug and all solvent fractions on body weight was determined. Blood samples was collected by cardiac puncture and retroorbital plexus method into EDTA sprinkled tubes and centrifuged at 3000 rpm for 20 min. Serum was separated as supernatant and stored at -20°C until analysis performed. The biochemical parameters, namely hemoglobin, urea, creatinine, total protein, total cholesterol, triglycerides, HDL determined. The other biochemical tests were performed using kits from Erba Diagnostics. Hesperidin and Apigenin significantly prevented the increase in blood glucose levels after 60 min. of glucose administration at the doses of 25 and 50 mg/kg. After 21 days blood samples were collected by retro-orbital plexus under mild anesthesia. Diabetic control group with no drug treatment showed no significant difference in the fasting serum glucose level after 21 days treatment as compared to the initial day treatment. Hesperidin & Apigenin (25 mg/kg & 50 mg/kg) treated diabetic rat showed a significant (p < 0.05) increase in body weight. The diabetic rat treated with glibenclaimide (10mg/kg) showed gradual and consistent increase in body weight. Treated diabetic rat with Hesperidin & Apigenin showed significant fall in total cholesterol, triglycerides, HDL-C, LDL-C and VLDL-C level at dose of 25 and 50 mg/kg. The Hesperidin and Apigenin had the antidiabetic activity with effects on lipids and other kidney markers..

Evaluating the Efficacy of Metformin and Camel Milk on Streptozotocin-nicotinamide Induced Diabetic Albino Rats

Background: This study was intended to evaluate the anti-diabetic activity of metformin and camel milk solely or in combination in streptozotocin- nicotinamide induced type II diabetic albino Wistar rats. Methods: Thirty albino Wistar rats were induced with type II diabetes through streptozotocin (60 mg/kg), followed by the administration of nicotinamide (120 mg/kg) after 15 min. The rats were randomly divided into control, diabetic control, metformin-treated (100 mg/kg), camel milk-treated (50 ml/rat/day) and metformin + camel milk-treated groups for 30 days. For this study, rats with blood glucose levels higher than 250 mg/dL were included. Blood glucose levels and body weight were measured at the beginning of the study and at regular intervals throughout the experiment.Furthermore, histopathological examination of liver, kidney and pancreatic tissue was conducted to determine the effects of metformin and camel milk on these organs. Result: The results showed that treatment with metformin, camel milk, or a combination of both significantly reduced fasting blood glucose levels and improved body weight compared to the diabetic control group. Morphological improvements were also observed in the histopathological examination of liver, kidney and pancreatic tissue. The results of this study are consistent with previous research showing the anti-diabetic effects of metformin and support the emerging role of alternative therapies such as camel milk. However, further research is needed to explore the potential mechanisms underlying the observed effects.

Investigation of preconditioning and the protective effects of nicotinamide against cerebral ischemia-reperfusion injury in rats

This study investigated the antioxidant and neuroprotective effects of nicotinamide combined with ischemic preconditioning against cerebral ischemia reperfusion (CIR) injury. Thirty-five Wistar albino male rats were randomly divided into five groups: sham, preconditioned ischemia/reperfusion (IP+IR), ischemia/reperfusion (IR), preconditioned ischemia/reperfusion + nicotinamide (IP+IR+N), and ischemia/reperfusion + nicotinamide (IR+N). CIR was achieved with bilateral common carotid artery occlusion. IP+IR and IP+IR+N groups 30 min before ischemia; Three cycles of 10 sec ischemia/30 sec reperfusion followed by 20 min IR were applied. The IP+IR+N and IR+N groups received 500 mg/kg nicotinamide intraperitoneally. After 24 h of reperfusion, a neurological evaluation was performed and vertıcal pole test. Biochemically, malondialdehyde (MDA), glutathione (GSH) levels and catalase (CAT) activity were measured in blood and brain tissue samples. Rates of red neurons, sateliosis and spongiosis were determined histopathologically in the prefrontal cortex areas.After CIR, MDA levels increased significantly in serum and brain tissue in the IR group compared to the sham group, while GSH and CAT activity decreased in the brain tissue (p < 0.05). MDA levels in the tissues were found significantly decreased in the IR+N group compared to the IR group (p < 0.05). Administration of nicotinamide together with IP significantly decreased MDA levels in brain tissue and increased GSH and CAT activity (p < 0.05). Compared to the IR group, the morphological and neurological damage in the prefrontal cortex areas decreased in the IP+IR, IP+IR+N, and IR+N groups (p < 0.05). In addition, red neuron, sateliosis and spongiosis rates increased significantly in the IR group compared to the Sham, IP+IR+N, IR+N groups (p < 0.001 for all). In neurological evaluation, while the neurological score increased and the time on the vertical pole decreased significantly in the IR group, preconditioning, and nicotinamide groups reversed (p < 0.05). The study’s results show that nicotinamide administration with ischemic preconditioning alleviates cerebral ischemia/reperfusion injury.

Therapeutic application of nicotinamide: As a potential target for inhibiting fibrotic scar formation following spinal cord injury.

We aimed to confirm the inhibitory effect of nicotinamide on fibrotic scar formation following spinal cord injury in mice using functional metabolomics. We proposed a novel functional metabolomics strategy to establish correlations between gene expression changes and metabolic phenotypes using integrated multi-omics analysis. Through the integration of quantitative metabolites analysis and assessments of differential gene expression, we identified nicotinamide as a functional metabolite capable of inhibiting fibrotic scar formation and confirmed the effect invivo using a mouse model of spinal cord injury. Furthermore, to mimic fibrosis models invitro, primary mouse embryonic fibroblasts and spinal cord fibroblasts were stimulated by TGFβ, and the influence of nicotinamide on TGFβ-induced fibrosis-associated genes and its underlying mechanism were examined. Administration of nicotinamide led to a reduction in fibrotic lesion area and promoted functional rehabilitation following spinal cord injury. Nicotinamide effectively downregulated the expression of fibrosis genes, including Col1α1, Vimentin, Col4α1, Col1α2, Fn1, and Acta2, by repressing the TGFβ/SMADs pathway. Our functional metabolomics strategy identified nicotinamide as a metabolite with the potential to inhibit fibrotic scar formation following SCI by suppressing the TGFβ/SMADs signaling. This finding provides new therapeutic strategies and new ideas for clinical treatment.

Exploring the Therapeutic Potential of Lepidagathis pungens Nees Extracts as Novel Therapy Options in Streptozotocin and Nicotinamide-Induced Diabetes in Albino Wistar Rats

Diabetes mellitus remains a global health concern, necessitating innovative therapeutic approaches. This study explores the antidiabetic potential of aqueous and methanol extracts from Lepidagathis pungens Nees in albino Wistar rats induced with diabetes. Diabetes induction involves the administration of streptozotocin (STZ) and nicotinamide (NIC), mimicking pathological conditions observed in human diabetes. The study employed aqueous and methanol extracts of L. pungens Nees alongside a standard drug, glibenclamide. Diabetes induction was achieved through intraperitoneal injection of STZ and NIC. Extracts and the standard drug were orally administered, and various parameters, including blood glucose levels, body weight changes, insulin and hemoglobin levels, as well as liver and lipid profiles, were monitored at specified intervals. Furthermore, liver and pancreas tissues were evaluated histopathologically. In comparison to the diabetic group, both aqueous and methanol extracts demonstrated significant antidiabetic activity (p &lt;0.001), with effects comparable to or better than the standard drug. A remarkable improvement in insulin and hemoglobin levels was observed, along with significant improvements in blood glucose levels and body weight changes. Furthermore, the extracts demonstrated positive effects on liver and lipid profiles, exhibiting potential to mitigate diabetes-induced complications. Histopathological studies revealed protective effects on liver and pancreas tissues, supporting the extracts’ therapeutic efficacy. In STZ- and NIC-induced diabetic rats, L. pungens Nees extracts show promising antidiabetic activity. The extracts demonstrated significant improvements in various physiological parameters, suggesting their potential as alternative or adjunctive therapies in the management of diabetes. Further research and clinical investigations are warranted to elucidate the underlying mechanisms and translate these findings into practical therapeutic applications.

Development of an Experimental Animal Screening Model for Induction of type-3 Diabetes Mellitus using High-fat Diet, Streptozotocin, and Nicotinamide

ABSTRACT Purpose: To develop a screening method for the induction of type-3 diabetes mellitus (T3DM) in experimental animals. Materials and Methods: Twelve Albino Wistar rats were divided into normal control (NC) and disease control (DC) group. Animals of NC were fed with high-fat diet (HFD) for four weeks followed by intraperitoneal administration of nicotinamide (230 mg/kg) and streptozotocin (STZ) (65 mg/kg). Body weight (BW) and blood glucose levels (BGL) were measured from week 1 to 4. Glycosylated hemoglobin (HbA1C) levels were estimated on week 4. Behavioral parameters including Morris water maze test (MWMT), T-maze test, and rota-rod test were recorded. Estimation of acetylcholine (ACh) and acetylcholinesterase (AChE) was conducted on week 4. Histopathological analysis of brain hippocampus was undertaken in three animals of each group. Results: DC group showed significant elevation in levels of BW and BGL with P value &lt; 0.05. Levels of HbA1C increased significantly in DC group with P value &lt; 0.0001. DC group showed a significant increase (P value &lt; 0.05 [week 1] and P value &lt; 0.001 [weeks 2–4]) in escape latency and elapsed time in MWMT and T-maze test, respectively. In DC group, elapsed time decreased significantly (P value &lt; 0.05 [week 1] and P value &lt; 0.001 [weeks 2–4]) in rota-rod test. A significant deviation in the levels of ACh and AChE was recorded with P value &lt; 0.0001. Histopathology of hippocampus part of DC group showed notable increase in the number of β-amyloid plaque. Conclusion: Animals administered with HFD, nicotinamide, and STZ showed dementia along with the diabetic condition suggesting the induction of T3DM.

Acute Combined Cerebrolysin and Nicotinamide Administration Promote Cognitive Recovery Through Neuronal Changes in the Hippocampus of Rats with Permanent Middle Cerebral Artery Occlusion

Stroke is one of the leading causes of disability worldwide, where the Hippocampus (HPC) is affected. HPC organizes memory, which is a cognitive domain compromised after a stroke, where cerebrolysin (CBL) and Nicotinamide (NAM) have been recognized as potentially therapeutic. In this study, we aimed to evaluate the efficacy of a combined administration of CBL and NAM in a rat stroke model. Male Sprague-Dawley rats (n = 36) were divided into four groups: saline (pMCAO − Saline), CBL (pMCAO + CBL), NAM (pMCAO + NAM), and experimental (pMCAO + CBL-NAM) (n = 9 per group). A permanent middle cerebral artery occlusion (pMCAO) was induced through electrocauterization of the middle cerebral artery, followed by the administration of CBL (2.5 ml/kg), NAM (500 mg/kg) or combined immediately after skin suture, as well as at 24, 48, and 72 h post-surgery. The rats were evaluated in the novel object recognition test; hippocampal infarct area measurement; reconstruction of neurons from CA1 for Sholl analysis; and, measurement of brain-derived neurotrophic factor (BDNF) levels near the infarct zone. Our findings revealed that the administration of CBL or NAM induced infarct reduction, improved cognition, and increased BDNF levels. Moreover, a combination of CBL and NAM increased dendritic intersection in CA1 pyramidal neurons. Thus, the combined administration of CBL and NAM can promote cognitive recovery after a stroke, with infarct reduction, cytoarchitectural changes in HPC CA1 neurons, and BDNF increase. Our findings suggest that this combination therapy could be a promising intervention strategy for stroke.

Hepatoprotective Effect of Extra Virgin Olive Oil and Apple Cider Vinegar in Type-2 Diabetic Rat Model

Introduction: Type-2 diabetes mellitus, a major public health problem, is caused by insulin resistance. In diabetics, chronic hyperglycemia results in formation of reactive oxygen species. Oxidative stress in diabetic patients is responsible for hepatic damage because it causes activation of several pro-inflammatory mediators in liver. Hepatic complications of diabetes include fatty liver disease, necrosis, cirrhosis, carcinoma and liver failure. Aims &amp; Objectives: To determine and compare the hepatoprotective effect of extra virgin olive oil and apple cider vinegar in streptozotocin induced Type-2 diabetic rat model. Place and Duration of Study: This experimental study was conducted at Animal House of Postgraduate Medical Institute, Lahore, Pakistan from May 2021 to June 2021. Material &amp; Methods: In this study, 40 male Sprague Dawley rats were divided into 4 groups i.e., Group I was NC (negative control), Group II PC (positive control), Group III EVOO (Extra virgin olive oil) and Group IV (Apple cider vinegar), each group having 10 rats. Diabetes was induced in all rats except the rats of NC group at the start of the study by intraperitoneal administration of injection nicotinamide, followed by injection STZ after 15 minutes. Group III was given 1ml/100gBW/ day EVOO and Group IV was given 2ml/kgBW/day diluted ACV with distilled water in 1:5 orally for 4 weeks. Terminal sampling was performed at the end of the 4th week for estimation of liver enzymes (ALT, ALP and AST) in serum. Data were entered and analyzed in SPSS version 26. p value of ? 0.05 was considered significant. Results: On comparison with PC group, both the treatment groups showed significant decrease in serum ALT levels with p values 0.026 for EVOO and &lt; 0.001 for ACV. Serum ALP levels were also decreased significantly in both the treated groups with p value &lt; 0.001 for EVOO as well as for the ACV group. However, reduction in serum AST levels was nonsignificant with p values 0.082 and 0.058 in EVOO and ACV groups respectively. Conclusion: Both EVOO and ACV have hepatoprotective effect in Type-2 diabetic rats. However, ACV is more effective.

Hypoxia modulation vs. chemotherapy and tumor shrinkage on early response assessment in diffuse large B-cell lymphoma

Background: Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) chemotherapy remains the standard of first-line treatment for diffuse large B-cell lymphoma (DLBCL). Up to 40% of DLBCL is characterized by relapse and refractory after treatment. Preliminary study reported Hypoxia-inducible factor-1? (HIF-1?) overexpression in 88.5% of DLBCL tumors in the Dr. Kariadi Hospital. Moreover, the role of hypoxia and HIF-1? has previously never been explored in DLBCL. Objectives: To evaluate the effect of hypoxia modulation to increased chemotherapeutic response in DLBCL. Methods: Single blind randomized control study was performed, with pre-test and post-test control group design. Research sampling consisted of DLBCL patients. The inclusion criteria include newly diagnosed DLBCL with HIF-1? overexpression and randomized to receive hypoxia modulation consisting of carbogen inhalation and nicotinamide administration, before R-CHOP chemotherapy. The tissue biopsy, histopathology and immunohistochemical studies were done. Chemotherapeutic responses were evaluated after 10-14 days following the first cycle of R-CHOP chemotherapy. Results: Out of twenty-six DLBCL participants with HIF-1? overexpression, there were 20 participants who completed the research protocol: 10 participants each in the intervention and control group. Demographic, clinicopathological, laboratory and disease characteristics were not statistically different between the two research groups (p&gt;0.05). Baseline tumor volume to be evaluated was also considered equal (172.3 cm3 vs. 152.8 cm3, p=0.597). Following the carbogen inhalation and nicotinamide administration, serum HIF-1? and lactate reduction can be observed. There was also a significant tumor volume shrinkage in both the intervention and control (mean ?85.7 cm3 vs. ?118.27 cm3) group, though the reduction was not statistically different (Delta 58.85% vs. 65.63%, p=0.474). Conclusion: The addition of hypoxia modulation to R-CHOP chemotherapy for DLBCL has shown beneficial effects on both serum HIF-1? and lactate concentration. However, the benefits did not correlate to increase a better tumor response compared to the control group.

92 Boosting in vitro bovine blastocyst production via intermittent administration of nicotinamide

92 Boosting in vitro bovine blastocyst production via intermittent administration of nicotinamide

Senescence and Mitochondrial Transfer Ability of Bone Marrow Mesenchymal Stromal Cells Influence Patterns of Alloreactivity after Allogeneic Hematopoietic Stem Cell Transplantation

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains the only curative treatment for acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). However post-transplant disease recurrence and alloreactive complications (chiefly graft versus host diseases - GvHD) are still associated with severe morbidity and mortality. Proliferative advantage of transformed cells as well as immune and microenvironmental dysregulations contribute both to promoting relapse and escape from graft versus leukemia (GvL) effect. Bone marrow (BM) mesenchymal stromal cells (MSC) are well known for their role in hematopoiesis and immune modulation. MSCs have been shown able to transfer mitochondria (MT) to different cell populations, including immune effectors, in order to (re)establish a healthy oxidative metabolism within the recipient cell. Here we hypothesize that these mechanisms are highly active in BM microenvironment after allo-HSCT and may contribute to alloreactivity, modifying the immune environment. On this basis we stipulate that those intercellular exchanges are deeply impacted by MSCs' senescence, that could be deployed as a useful parameter to predict post-transplant outcomes. To that end, we conducted a translational research, based on BM specimens prospectively collected at our Institution (NCT03357172, NCT03964922), involving patients undergoing allo-HSCT for high-risk MDS and AML and receiving fludarabine/busulfan-based conditioning regimens, anti-lymphoglobuline and cyclosporin/mycophenolate mofetil. MSC phenotype and functionality at the pre-transplant stage, MT capacity, immunomodulation towards T cells and phenotype were studied by multiparametric flow cytometry. Cell metabolism, in terms of oxygen consumption and extracellular acidification rate, was characterized with Seahorse XF technology. Percentages of senescent MSCs were determined with SA β-galactosidase test. First, we compared phenotypic and metabolic characteristics of MSC from MDS/AML (N=29) patients with those retrieved from a cohort of healthy controls (N=8). We observed that patient MSCs were characterized by higher expression of CD157 and CD146 and lower expression of CD200 as compared to healthy MSCs. Pre-transplant senescence quantification highlighted a certain degree of heterogeneity within the MSC compartment in leukemic patients (median = 37.42% sd=16.30 for AML patient and m=11.74% sd=6.4 for HC). In an attempt to link clinical phenotypes to this heterogeneity, we observed that patients with high degree of MSC senescence in pretransplant samples (mean= 51.6% sd=16.6 cells/sample), developed more frequently acute GvHD, as compared to those with low degree of pre-transplant MSC senescence (mean= 23.8% sd=14.3) who experienced instead a higher incidence of disease relapse (p=0,004). Assuming that the link between senescence and alloreactivity could rely to different metabolic states and MT capabilities in MSCs, we analyzed these biological features in our samples. Indeed, we observed that low degree of MSC senescence was associated with higher MT to T cells, in particular to CD4+ cells, shifting their metabolism and phenotype toward an immunosuppressive state The characterization of cellular metabolism showed in highly senescent MSCs the presence of markers of mitochondrial dysfunction with high oxidative power, but reduced coupling efficiency of respiratory chain. This metabolic profile involved a high mitochondrial mass characterized by a high maximal respiration capacity compared to HC MSCs. The in vitro modulation of MSC senescence with nicotinamide administration restored healthy mitochondrial metabolism and improved coupling efficacy of oxidative phosphorylation, possibly impacting on the immunomodulation capabilities of this microenvironment component. Here for the first time, we provide the evidence that MSC senescence plays an pivotal role in defining an altered immune state in transplanted AML possibly shifting the degree of alloreactivity from a high GvH/GvL response (high GvHD and low relapse) to a high immunosuppressive state (low GvHD and high relapse). These findings pave the way for implementing MSC senescence as a biomarker able to predict post-transplant outcomes and show new biological rationales to integrate donor-derived MSC products to better adjust GvH responses.

Astaxanthin attenuates cognitive deficits in Alzheimer’s disease models by reducing oxidative stress via the SIRT1/PGC-1α signaling pathway

ObjectiveOxidative stress plays a pivotal role in neurodegenerative diseases. Astaxanthin (AST) can play a neuroprotective role owing to its long-chain conjugated unsaturated double bond, which imparts potent antioxidant, anti-neuroinflammatory, and anti-apoptotic properties. However, the biological mechanisms underlying these effects remain unknown. Therefore, this study aimed to investigate and validate the protective effect of AST on neuronal senescence and apoptosis caused by oxidative stress induced by Aβ25–35 peptide, with the goal of preventing the onset of cognitive dysfunction.MethodsAlzheimer's disease models comprising ICR mice and PC12 cells were established using Aβ25–35. The Morris water maze test was used to assess mouse behavior. Nissl staining revealed morphological changes in the mouse hippocampal neurons. To elucidate the mechanism of action of AST, ICR mice and PC12 cells were treated with the silent information regulator 1 (SIRT1) inhibitor nicotinamide (NAM). Additionally, immunofluorescence, western blotting, and reverse transcription polymerase chain reaction were used to evaluate changes in the expression of Bcl-2 and Bax in the mouse hippocampus, and SIRT1/PGC-1α signaling pathway proteins were detected. Moreover, the oxidative stress markers in ICR mice and PC12 cells were evaluated. Further, CCK-8 assays, Annexin V/PI double staining, and β-galactosidase activity assays were performed in PC12 cells to evaluate the anti-senescence and apoptotic effects of AST.ResultsIn vivo experiments showed that Aβ25–35 impaired cognitive function, promoted morphological changes in hippocampal neurons, decreased Bcl-2 expression, increased Bax expression, decreased superoxide dismutase and GSH-px levels, and increased reactive oxygen species and malondialdehyde levels. Conversely, AST alleviated the impact of Aβ25–35 in mice, with reversed outcomes. NAM administration reduced SIRT1 and PGC-1α expression in the hippocampus. This decrease was accompanied by cognitive dysfunction and hippocampal neuron atrophy, which were also evident in the mice. Additionally, in vitro experiments showed that Aβ25–35 could promote oxidative stress and induce the senescence and apoptosis of PC12 cells. Nonetheless, AST treatment counteracted this effect by inhibiting oxidative stress and altering the state of PC12 cells. Notably, the Aβ + NAM group exhibited the most significant rates of senescence and apoptosis in PC12 cells following NAM treatment.ConclusionAST can improve cellular senescence and apoptosis mediated by oxidative stress via the SIRT1/PGC-1α signaling pathway and plays a vital role in inhibiting neuronal senescence and apoptosis and enhancing cognitive ability.

Pterostilbene ameliorates type-2 diabetes mellitus – Induced depressive-like behavior by mitigating insulin resistance, inflammation and ameliorating HPA axis dysfunction in rat brain

One of the most serious mental health comorbidities associated with diabetes mellitus is depression. The occurrence is almost double in type 2 diabetes mellitus (T2DM) compared with the general population. Pterostilbene (PTE), a dimethylated analog of resveratrol, has been reported for significant neuroprotective, anti-inflammatory, hypolipidemic, hypoglycaemic and antioxidant effects. However, its effect on diabetes-induced depression-like behavior (DID) has not been studied.The current study aimed at studying the effects of PTE on depressive-like behavior in male Wistar rats with T2DM. It was induced by single dose administration of nicotinamide (NA) and streptozotocin (STZ). On day 21, forced swim test (FST) was conducted for the confirmation of DID. Rats demonstrating depressive-like behavior were treated with PTE (10, 20 and 40 mg/kg), metformin (MET; 500 mg/kg) and fluoxetine (FLX; 10 mg/kg) for 28 days, orally. At the end of the treatment, behavioral assessment for depression, blood glucose (BG) and lipid profile, oxidative stress markers, gene expression of Sirtuin 1 (SIRT1) and histopathological parameters were investigated. PTE significantly reduced weight loss and mitigated depressive-like behavior paradigms such as sucrose preference test (SPT), resident intruder test (RIT) and open field test (OFT). It significantly restored BG, lipid and liver profile, creatinine and antioxidant level. It Improved glucose tolerance, insulin resistance (IR) and reduced cortisol level as well as inflammatory markers. It showed improved morphology of the pancreas, brain, liver and kidney. Gene expression studies revealed that, PTE significantly increased SIRT1 expression. PTE by its virtue to maintain BG, reduced IR, amelioration of the HPA axis, anti-inflammatory, antioxidant activity and improvement of SIRT1 gene expression proved to be effective in the treatment of DID-like behavior in rats.

Comparative analysis of the pharmacokinetic parameters of transdermal and injectable forms of nicotinamide

In recent years, oxidative stress, characterized by excess free radicals in the body, has been called the cause of many diseases. There is an active search for drugs with antioxidant properties that are suitable for long-term maintenance therapy. Nicotinamide (NAM), an antioxidant, is used to treat a variety of diseases, usually in oral or injectable form. Given the peculiarities of the drug regimen (dose, prolonged administration), a new dosage form of NAM, a microemulsion-based transdermal patch (TP), containing 20 mg/10 cm2 of NAM, has been proposed.The objective of this work is to compare the pharmacokinetic parameters of intramuscular and transdermal NAM administration in animal experiments for 24 hours.Materials and methods. We used laboratory samples of nicotinamide TP based on a microemulsion-based transdermal delivery emulsion (TDS) with different content of sodium docusate transfer activator. The pharmacokinetics of transdermal and intramuscular injections were studied in male Chinchilla rabbits weighing 3.5–4.0 kg. Plasma NAM levels of the experimental animals were determined by high-performance liquid chromatography using a specially designed method on NUCLEODUR PFP columns (5 μm, 250 × 4.6 mm) using the mobile phase acetonitrile: deionized water. The samples were preliminarily purified by solid-phase extraction using Chromabond C18 Hydra cartridges.Results. When administered intramuscularly, the maximum blood NAM level was 13.3±1 μg/mL; when NAM transdermal forms were applied in the same dosage with different contents of the transfer activator, the levels did not differ significantly – 3.1 and 3.2 μg/mL. It was shown that in transdermal administration of NAM, concentration of the active substance remained at a constant level for ~6 hours. The bioavailability of NAM with transdermal administration was calculated relative to intramuscular administration: 1.43 for TP with 9.8% docusate sodium and 1.84 with 3.3% docusate sodium.Conclusion. NAM has a higher bioavailability when administered transdermally at 20 mg than when administered intramuscularly in the same dose. With transdermal administration, NAM concentration can be maintained at a constant level for a long time, without the jumps that are typical of intramuscular administration.

Neuroprotective effects of terpenoids against streptozotocin-nicotinamide-induced diabetic rats: An in silico, in vitro and in vivo study

The present study focused to determine the neuroprotective effects of terpenoids in streptozotocin & nicotinamide-induced type-2 diabetes in rats. The in silico studies were carried out for 68 terpenoids using AutoDock 4.2. The in vitro cholinestrerase, α-amylase enzyme inhibitory assays were perfomed using standard procedures. For in vivo neuroprotective studies, male wistar rats were separated into five groups and each group comprised of six animals. Treatment groups were received low dose and high dose α-Bisabolol 100 and 200 mg/kg respectively, and the standard groups received rivastigmine 2 mg/kg, p.o. and metformin group 100 mg/kg, p.o. for 30 consecutive days. Administration of streptozotocin (45 mg/kg, i.p.) and nicotinamide (110 mg/kg, i.p.)-induced the type 2 diabetes in all groups except the control. The behavioural assessments such as Morris water maze, and open field test were performed and biochemical parameters such as acetylcholinesterase levels and enzymatic antioxidants and reduced glutathione level were estimated from brain homogenates. Treatment of diabetic rats with α-Bisabolol was lowered blood glucose level, improved spatial recognition memory in behavioural assessments in a concentration dependent manner. It can be concluded that α-Bisabolol could act as a potential drug candidate in the management of diabetic Alzheimer's disease.

Nicotinamide ameliorates mitochondria-related neuronal apoptosis and cognitive impairment via the NAD+/SIRT3 pathway

Emerging evidence suggests that mitochondria play a central role in mental health disorders including schizophrenia. Here we investigated whether nicotinamide (NAM) normalized cognitive impairment via a mechanism involving the mitochondrial Sirtuin 3 (SIRT3) pathway. The 24 h maternal separation (MS) rat model was used to mimic schizophrenia-associate phenotypes. Schizophrenia-like behaviors and memory impairments were detected using the pre-pulse inhibition test, novel object recognition test, and Barnes maze test, and neuronal apoptosis was characterized using multiple assays. SIRT3 activity was inhibited pharmacologically or by knockdown in HT22 cells, and BV2 microglia and SIRT3-knockdown HT22 cells were co-cultured in vitro. Mitochondrial molecules were measured by western blotting, and mitochondrial damage was measured with reactive oxygen species and mitochondrial membrane potential assays. Proinflammatory cytokines were assayed by ELISA and microglial activation was detected by immunofluorescence. MS animals showed behavioral and cognitive impairment and increased neuronal apoptosis. Supplementation with NAM or administration of honokiol, a SIRT3 activator, reversed all of the changes in behavioral and neuronal phenotypes. Administration of the SIRT3 inhibitor 3-TYP in control and NAM-treated MS rats caused behavioral and neuronal phenotypes similar to MS. In vitro, inhibition of SIRT3 activity with 3-TYP or by knockdown in HT22 cells increased ROS accumulation and caused neuronal apoptosis in a single-culture system. In co-culture systems, SIRT3 knockdown in HT22 cells activated BV2 microglia and increased levels of TNF-α, IL-6, and IL-1β. The administration of NAM blocked these alterations. Taken together, these data suggest that NAM can rescue neuronal apoptosis and microglial over-activation through the nicotinamide adenine dinucleotide (NAD+)-SIRT3-SOD2 signaling pathway, furthering our understanding of the pathogenesis of schizophrenia and providing avenues for novel treatments.

Therapeutic investigations of palm oil induced beta-carotene in diabetic vascular dementia in rat

Vascular dementia (VaD) is a major factor for the progress of stroke and other memory disorders. It is the second leading cause of death. The burden of VaD is higher in the aged population (&gt;65 years). The progression of VaD occurs with lifestyle modifications i.e., fast food, smoking, and alcohol. The medicines for the treatment of VaD are limited. The palm oil is one of the rich sources of beta-carotene (BC). The present research designed to investigate the potential role of palm oil mill effluent derived BC in experimental model of diabetic VaD. The diabetic VaD was induced by administration of nicotinamide (NA, 50 mg/kg; i.p.) followed by streptozotocin (STZ, 50 mg/kg; i.p.). The test compound i.e., BC (50 and 100 mg/kg) and reference compound donepezil (1 mg/kg) were administered orally for 15 consecutive days. The changes of cognitive patterns i.e., escape latency time (ELT) and time spent in target quadrant (TSTQ) was assessed by Morris water maze (MWM) test. Besides the changes of neurotransmitter i.e., acetylcholinesterase (AChE) was estimated in brain (hippocampus, cerebellum, entorhinal cortex, amygdala and septum) samples. The administration of STZ caused the significant changes of cognitive functions (increased ELT and decreased TSTQ) as indicated in the development of VaD when compared to normal group. The treatment of BC was ameliorated the cognitive dysfunctions against the STZ associated cholinergic neurotransmitter (elevated AChE) changes. The effects were similar to donepezil treatment group. Hence, it proved that BC possesses the potential therapeutic effects in the management of diabetic VaD due to its potential anti-cholinergic effects.