AIMS: This study investigates the neuroprotective effects of lipidized analogues of 2-SS-CART(61-102) derived from anorexigenic neuropeptide cocaine- and amphetamine-regulated transcript peptide (CARTp) in light of the link between obesity, its comorbidities, and the development of Alzheimer's disease. METHODS: We introduce novel lipidized analogues derived from 2-SS-CART(61-102), a specific analogue of natural CART(61-102), with two disulfide bridges. Using hypothermic PC12 cells, we tested the effect of the most potent analogues on Tau phosphorylation. We further described the anorexigenic and neuroprotective potential of subcutaneously (SC) injected lipidized CARTp analogue in a mouse model with prediabetes and obesity induced by neonatal monosodium glutamate (MSG) administration. RESULTS: Compared to the non-lipidized 2-SS-CART(61-102), all lipidized analogues exhibited a potent binding affinity to PC12 cells and enhanced in vitro stability in rat plasma. Two most potent lipidized analogues attenuated hypothermia-induced Tau hyperphosphorylation at multiple epitopes. Subsequently, chronic SC treatment with palm-2-SS-CART(61-102) significantly decreased body weight and food intake, improved metabolic parameters, decreased level of pTau and increased neurogenesis in hippocampi of obese MSG mice. CONCLUSION: Our unique CARTp analogue palm-2-SS-CART(61-102) shows promise as a potent anti-obesity and neuroprotective agent.

• MeSH
• Appetite Depressants pharmacology   MeSH
• PC12 Cells MeSH
• Phosphorylation drug effects   MeSH
• Sodium Glutamate * MeSH
• Rats MeSH
• Lipids chemistry   blood   MeSH
• Disease Models, Animal MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Neuroprotective Agents * pharmacology   MeSH
• Obesity * metabolism   drug therapy   MeSH
• Nerve Tissue Proteins * metabolism   MeSH
• tau Proteins metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Although hypothermic treatment has been reported to have some beneficial effects on ischaemia at the clinical level, the mechanism of ischaemia suppression by hypothermia remains unclear due to a lack of mechanism understanding and insufficient data. The aim of this study was to isolate and characterize microRNAs specifically expressed in ischaemia-hypothermia for the dihydropyrimidinase-like 3 (Dpysl3) gene. PC12 cells were induced with CoCl2 for chemical ischaemia and incubated at 32 °C for hypothermia. In ischaemia-hypothermia, four types of microRNAs (miR-106b-5p, miR-194-5p, miR-326-5p, and miR-497-5p) were highly related to the Dpysl3 gene based on exosomal microRNA analysis. Dpysl3 gene expression was up-regulated by miR-497-5p but down-regulated by miR-106b-5p, miR-194-5p and miR-326-5p. Our results suggest that these four microRNAs are involved in the regulation of Dpysl3 gene expression. These findings provide valuable clues that exosomal microRNAs could be used as therapeutic targets for effective treatment of ischaemia.

• MeSH
• PC12 Cells MeSH
• Gene Expression MeSH
• Hypothermia * genetics   MeSH
• Ischemia MeSH
• Rats MeSH
• Humans MeSH
• MicroRNAs * genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Labile redox-active iron ions have been implicated in various neurodegenerative disorders, including the Parkinson's disease (PD). Iron chelation has been successfully used in clinical practice to manage iron overload in diseases such as thalassemia major; however, the use of conventional iron chelators in pathological states without systemic iron overload remains at the preclinical investigative level and is complicated by the risk of adverse outcomes due to systemic iron depletion. In this study, we examined three clinically-used chelators, namely, desferrioxamine, deferiprone and deferasirox and compared them with experimental agent salicylaldehyde isonicotinoyl hydrazone (SIH) and its boronate-masked prochelator BSIH for protection of differentiated PC12 cells against the toxicity of catecholamines 6-hydroxydopamine and dopamine and their oxidation products. All the assayed chelating agents were able to significantly reduce the catecholamine toxicity in a dose-dependent manner. Whereas hydrophilic chelator desferrioxamine exerted protection only at high and clinically unachievable concentrations, deferiprone and deferasirox significantly reduced the catecholamine neurotoxicity at concentrations that are within their plasma levels following standard dosage. SIH was the most effective iron chelator to protect the cells with the lowest own toxicity of all the assayed conventional chelators. This favorable feature was even more pronounced in prochelator BSIH that does not chelate iron unless its protective group is cleaved in disease-specific oxidative stress conditions. Hence, this study demonstrated that while iron chelation may have general neuroprotective potential against catecholamine auto-oxidation and toxicity, SIH and BSIH represent promising lead molecules and warrant further studies in more complex animal models.

• MeSH
• PC12 Cells MeSH
• Iron Chelating Agents * pharmacology   MeSH
• Deferasirox pharmacology   MeSH
• Deferiprone pharmacology   MeSH
• Deferoxamine pharmacology   MeSH
• Dopamine pharmacology   MeSH
• Catecholamines pharmacology   MeSH
• Rats MeSH
• Oxidative Stress MeSH
• Oxidopamine pharmacology   MeSH
• Iron Overload * MeSH
• Iron pharmacology   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Metal-organic frameworks (MOFs) have been widely used as porous nanomaterials for different applications ranging from industrial to biomedicals. An unpredictable one-pot method is introduced to synthesize NH2-MIL-53 assisted by high-gravity in a greener media for the first time. Then, porphyrins were deployed to adorn the surface of MOF to increase the sensitivity of the prepared nanocomposite to the genetic materials and in-situ cellular protein structures. The hydrogen bond formation between genetic domains and the porphyrin' nitrogen as well as the surface hydroxyl groups is equally probable and could be considered a milestone in chemical physics and physical chemistry for biomedical applications. In this context, the role of incorporating different forms of porphyrins, their relationship with the final surface morphology, and their drug/gene loading efficiency were investigated to provide a predictable pattern in regard to the previous works. The conceptual phenomenon was optimized to increase the interactions between the biomolecules and the substrate by reaching the limit of detection to 10 pM for the Anti-cas9 protein, 20 pM for the single-stranded DNA (ssDNA), below 10 pM for the single guide RNA (sgRNA) and also around 10 nM for recombinant SARS-CoV-2 spike antigen. Also, the MTT assay showed acceptable relative cell viability of more than 85% in most cases, even by increasing the dose of the prepared nanostructures.

• MeSH
• Hep G2 Cells MeSH
• PC12 Cells MeSH
• COVID-19 diagnosis   MeSH
• CRISPR-Cas Systems MeSH
• Nitrogen chemistry   MeSH
• RNA, Guide, Kinetoplastida MeSH
• HEK293 Cells MeSH
• HeLa Cells MeSH
• DNA, Single-Stranded MeSH
• Rats MeSH
• Humans MeSH
• Limit of Detection MeSH
• Nanocomposites MeSH
• Nanostructures MeSH
• Metal-Organic Frameworks chemistry   MeSH
• Porosity MeSH
• Porphyrins chemistry   MeSH
• Surface Properties MeSH
• RNA, Viral metabolism   MeSH
• SARS-CoV-2 MeSH
• Sensitivity and Specificity MeSH
• COVID-19 Testing MeSH
• Hydrogen Bonding MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

CART (cocaine- and amphetamine-regulated transcript) peptides are involved in food intake regulation, stress, and other physiological functions. Although CART peptides have been known for over 25 years, their receptor(s) have not yet been characterized. In this short review, we will summarize our previous studies, where we reported specific binding of 125 I-CART(61-102) to PC12 rat pheochromocytoma cells. Competitive binding experiments performed with mono- and di-iodinated peptides and their isoforms with oxidized Met67 resulted in nanomolar binding affinity. Moreover, in our previous study, CART(61-102), as well as di-iodinated CART(61-102), have shown a strong anorexigenic effect in fasted lean mice after intracerebroventricular administration. In conclusion, from our previous studies, iodination of CART(61-102) resulted in mono- and di-iodinated analogs with or without oxidized Met67 . All analogs revealed a high affinity to binding sites at PC12 cells and preserved biological activity.

• MeSH
• Appetite Depressants chemistry   pharmacokinetics   therapeutic use   MeSH
• PC12 Cells MeSH
• Rats MeSH
• Mice MeSH
• Nerve Tissue Proteins chemistry   pharmacokinetics   therapeutic use   MeSH
• Radiopharmaceuticals chemistry   pharmacokinetics   therapeutic use   MeSH
• Iodine Radioisotopes chemistry   MeSH
• Protein Binding MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

Reperfusion therapies for ischaemic stroke can induce secondary injury accompanied by neuronal death. The Y-box binding protein 1 (YBX1), an oncoprotein, is critical for regulating tumour cell proliferation and apoptosis. Thus, we wanted to know whether YBX1 could regulate neuronal cell apoptosis caused by cerebral ischaemia/reperfusion (I/R). We established a model of cerebral I/R-induced injury in vitro by oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and determined YBX1 expression using Western blot. Next, the effect of YBX1 on the apoptosis and viability of OGD/R-treated PC12 cells was evaluated by flow cytometry, MTT assay, and Western blot. Besides, the release of lactate dehydrogenase (LDH) and the activity of catalase (CAT) and superoxide dismutase (SOD) were detected to evaluate oxidative stress of PC12 cells induced by OGD/R. The regulatory roles of YBX1 in the AKT/GSK3β pathway were examined by Western blot. As a result, OGD/R treatment down-regulated YBX1 expression in PC12 cells. YBX1 over-expression attenuated the growth inhibition and apoptosis of PC12 cells induced by OGD/R. Besides, the increase of LDH release and the decrease of SOD and CAT activities caused by OGD/R were reversed by YBX1 over-expression. Moreover, YBX1 over-expression could activate the AKT/GSK3β pathway in OGD/ R-treated PC12 cells. Therefore, YBX1 could protect against OGD/R-induced injury in PC12 cells through activating the AKT/GSK3β signalling pathway, and thus YBX1 has the potential to become a therapeutic target for cerebral I/R-induced injury.

• MeSH
• Apoptosis MeSH
• PC12 Cells MeSH
• Stroke * MeSH
• DNA-Binding Proteins MeSH
• Glucose MeSH
• Brain Ischemia * MeSH
• Glycogen Synthase Kinase 3 beta * MeSH
• Rats MeSH
• Oxygen MeSH
• Oxidative Stress MeSH
• Proto-Oncogene Proteins c-akt * MeSH
• Reperfusion Injury * MeSH
• Cell Survival MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

PURPOSE: The aim of this study is to show a new mesomicroscopic insight into Letrozole (LTZ) loaded nanocomplexes and their ex vivo characteristics as a drug delivery system. METHODS: The LTZ loaded hybrid chitosan-based carrier was fabricated using a modified ionic crosslinking technique and characterized in more detail. To understand the mechanism of LTZ action encapsulated in the hybrid polymer-lipid carrier, all-atom molecular dynamics simulations were also used. RESULTS: The physicochemical properties of the carrier demonstrated the uniform morphology, but different drug loading ratios. In vitro cytotoxic activity of the optimized carrier demonstrated IC50 of 67.85 ± 0.55 nM against breast cancer cell line. The ex vivo study showed the positive effect of nanocomplex on LTZ permeability 7-10 fold greater than the free drug. The molecular dynamic study also confirmed the prsence of hydrophobic peak of lipids at a distance of 5 Å from the center of mass of LTZ which proved drug entrapment in the core of nanocomplex. CONCLUSIONS: The hybrid nanoparticle increased the cytotoxicity and tissue permeability of LTZ for oral delivery. This study also confirmed the atomic mesostructures and interaction of LTZ in the core of hybrid polymer-lipid nanoparticles.

• MeSH
• PC12 Cells MeSH
• Chitosan chemistry   MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Rats MeSH
• Drug Delivery Systems methods   MeSH
• Letrozole chemistry   MeSH
• Humans MeSH
• Lipids chemistry   MeSH
• MCF-7 Cells MeSH
• Cell Line, Tumor MeSH
• Nanoparticles chemistry   MeSH
• Drug Carriers chemistry   MeSH
• Polymers chemistry   MeSH
• Molecular Dynamics Simulation MeSH
• Particle Size MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Neuroinflammation and cholinergic deficit are key detrimental processes involved in Alzheimer's disease. Hence, in the search for novel and effective treatment strategies, the multi-target-directed ligand paradigm was applied to the rational design of two series of new hybrids endowed with anti-inflammatory and anticholinesterase activity via triple targeting properties, namely able to simultaneously hit cholinesterases, cyclooxygenase-2 (COX-2) and 15-lipoxygenase (15-LOX) enzymes. Among the synthesized compounds, triazoles 5b and 5d, and thiosemicarbazide hybrid 6e emerged as promising new hits, being able to effectively inhibit human butyrylcholinesterase (hBChE), COX-2 and 15-LOX enzymes with a higher inhibitory potency than the reference inhibitors tacrine (for hBChE inhibition), celecoxib (for COX-2 inhibition) and both NDGA and Zileuton (for 15-LOX inhibition). In addition, compound 6e proved to be a submicromolar mixed-type inhibitor of human acetylcholinesterase (hAChE). The anti-neuroinflammatory activity of the three most promising hybrids was confirmed in a cell-based assay using PC12 neuron cells, showing decreased expression levels of inflammatory cytokines IL-1β and TNF-α. Importantly, despite the structural resemblance to tacrine, they showed ideal safety profiles on hepatic and murine brain cell lines and were safe up to 100 μM when assayed in PC12 cells. All three hybrids were also predicted to have superior BBB permeability than tacrine in the PAMPA assay, and good physicochemical properties, drug-likeness and ligand efficiency indices. Finally, molecular docking studies highlighted key structural elements impacting selectivity and activity toward the selected target enzymes. To the best of our knowledge, compounds 5b, 5d and 6e are the first balanced, safe and multi-target compounds hitting the disease at the three mentioned hubs.

• MeSH
• Acetylcholine deficiency   MeSH
• Alzheimer Disease drug therapy   pathology   MeSH
• Cell Line MeSH
• PC12 Cells MeSH
• Cholinesterase Inhibitors chemistry   MeSH
• Cyclooxygenase 2 Inhibitors chemistry   MeSH
• Lipoxygenase Inhibitors chemistry   MeSH
• Rats MeSH
• Humans MeSH
• Mice MeSH
• Neurons drug effects   enzymology   pathology   MeSH
• Drug Design MeSH
• Semicarbazides chemistry   pharmacology   MeSH
• Molecular Docking Simulation MeSH
• Triazoles chemistry   pharmacology   MeSH
• Inflammation drug therapy   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Ischemic stroke is a severe cause of disability and death all over the world. To search for effective therapy for ischemic stroke, PC12 cells damaged by oxygenation and glucose deprivation/restoration were employed to assess the protective effects of inotodiol. As a result, inotodiol can improve the cell viability and attenuate the leakage of lactate dehydrogenase. Meanwhile, inotodiol can prevent oxidative stress by reducing reactive oxygen species generation, decreasing the content of malonic dialdehyde, and increasing the activity of superoxide dismutase. In addition, the dysfunction of mitochondria induced by oxygenation and glucose deprivation/restoration was ameliorated through decreasing the level of intracellular calcium and increasing the mitochondrial membrane potential. At the same time, inotodiol can inhibit PC12 cells apoptosis through downregulation of Caspase-3 and Bax as well as upregulation of Bcl-2. These results reveal inotodiol can protect PC12 cells against the injury induced by oxygenation and glucose deprivation/restoration. This investigation gives promising evidences for the therapy of ischemic stroke.

• Keywords
• inotodiol,
• MeSH
• Apoptosis MeSH
• PC12 Cells * pathology   drug effects   MeSH
• Stroke * drug therapy   MeSH
• Glucose MeSH
• Caspase 3 MeSH
• Rats MeSH
• Oxygen MeSH
• Lactate Dehydrogenases metabolism   MeSH
• Lanosterol analogs & derivatives   pharmacology   MeSH
• Membrane Potential, Mitochondrial drug effects   MeSH
• Oxidative Stress drug effects   MeSH
• Reactive Oxygen Species metabolism   MeSH
• Calcium Signaling drug effects   MeSH
• Blotting, Western MeSH
• Check Tag
• Rats MeSH

Alzheimer's disease (AD) threatens to become the scourge of the 21st century, hence there is an urgent need for the exploration of multipotent drug in the treatment of AD. Rhizophora mucronata, tropical red mangrove has been widely used in traditional oriental medicine as astringent, antiseptic, antibacterial, anti-ulcerogenic and anti-inflammatory agent. The present study aimed to evaluate the neuroprotective effect of the catechin rich methanolic leaf extract of R. mucronata (MERM), against Aβ-induced neurotoxicity. Exposure of PC12 cells to Aβ (25–35) increased cellular oxidative stress, the number of apoptotic cells and caspase-3 activity ultimately leading to neuronal death. Pre-treatment with MERM (50 μg/ml) significantly attenuated cell death, decreased the level of intracellular reactive oxygen and nitrogen species thereby inhibiting lipid peroxidation and protein oxidation. MERM attenuated Aβ (25–35) induced apoptosis by stabilizing the mitochondrial membrane potential, and inhibiting caspase-3 activity. MERM also restored the antioxidant status of the Aβ (25–35) treated cells by effectively scavenging ROS/RNS species. HPTLC analysis of MERM illustrated the presence of (+)-catechin as major constituent. Results conclude that MERM effectively attenuated the neurotoxicity induced by Aβ-associated oxidative stress, implying that MERM can act as a potent drug for the treatment of AD.

• MeSH
• Alzheimer Disease drug therapy   MeSH
• Amyloid beta-Peptides * drug effects   MeSH
• Apoptosis MeSH
• PC12 Cells * drug effects   MeSH
• Chromatography, Thin Layer MeSH
• Catechin MeSH
• Cells, Cultured MeSH
• Plant Leaves MeSH
• Membrane Potential, Mitochondrial MeSH
• Neuroprotective Agents pharmacology   chemical synthesis   chemistry   MeSH
• Oxidative Stress MeSH
• Peptide Fragments drug effects   MeSH
• Reactive Oxygen Species MeSH
• Rhizophoraceae * MeSH
• Plant Extracts MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH