1-palmitoyl-2-oleoylphosphatidylcholine and Disease-Models--Animal

Treatment of

Topics: Animals; Anti-Bacterial Agents; Bithionol; Cell Membrane; Cell Membrane Permeability; Cholesterol; Disease Models, Animal; Drug Repositioning; Drug Synergism; Gentamicins; Lipid Bilayers; Membrane Fluidity; Methicillin-Resistant Staphylococcus aureus; Molecular Dynamics Simulation; Phosphatidylcholines; Structure-Activity Relationship; Unilamellar Liposomes

64Cu radiolabelled nanodiscs based on the 11 α-helix MSP1E3D1 protein and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine lipids were, for the first time, followed in vivo by positron emission tomography for evaluating the biodistribution of nanodiscs. A cancer tumor bearing mouse model was used for the investigations, and it was found that the approximately 13 nm nanodiscs, due to their size, permeate deeply into cancer tissue. This makes them promising candidates for both drug delivery purposes and as advanced imaging agents. For the radiolabelling, a simple approach for 64Cu radiolabelling of proteins via a chelating agent, DOTA, was developed. The reaction was performed at sufficiently mild conditions to be compatible with labelling of the protein part of a lipid-protein particle while fully conserving the particle structure including the amphipathic protein fold.

Topics: Animals; Cell Line, Tumor; Copper Radioisotopes; Disease Models, Animal; Heterocyclic Compounds, 1-Ring; Heterografts; Humans; Mice; Nanostructures; Neoplasms; Particle Size; Phosphatidylcholines; Positron-Emission Tomography; Radiopharmaceuticals; Tissue Distribution; Tomography, X-Ray Computed

The present study investigated the effects of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DL-PC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PO-PC) on depression-related behaviors and spatial memory impairment in mice subjected to restraint stress. The immobility time in forced-swim and tail-suspension tests for mice subjected to restraint stress was significantly longer than that for nonstressed control mice, and oral coadministration of DL-PC and PO-PC (DL-/PO-PC; DL-PC : PO-PC=1 : 1) shortened the prolonged immobility time in a dose (0.1-5 mg/kg)-dependent manner. In the water maze test, the retention latency for stressed mice was significantly longer than that for control mice and DL-/PO-PC (1 mg/kg, per os) reversed the prolonged latency to control levels. Phosphorylation of Akt and glycogen synthase kinase 3β (GSK-3β) in the hypothalamus of stressed mice was significantly reduced compared with that for control mice, and DL-/PO-PC (1 mg/kg, per os) recovered the reduced phosphorylation of Akt and GSK-3β. The results of the present study indicate that DL-/PO-PC has the potential to ameliorate stress-induced depression-related behaviors and memory impairment, possibly by activating Akt and inhibiting GSK-3β.

Topics: Animals; Depressive Disorder; Disease Models, Animal; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Hypothalamus; Male; Maze Learning; Memory Disorders; Mice, Inbred C57BL; Motor Activity; Neuropsychological Tests; Phosphatidylcholines; Phosphorylation; Proto-Oncogene Proteins c-akt; Psychotropic Drugs; Restraint, Physical; Spatial Memory; Stress, Psychological

In the water maze test, oral administration with 1,2-dilynoleoyl-sn-glycero-3-phosphocholine (DLPhtCho)(5 mg/kg) alone or DLPhtCho (5 mg/kg) plus 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPhtCho)(5 mg/kg) significantly shortened the prolonged acquisition latency for rats intraperitoneally injected with scopolamine, with more efficient effect than (POPhtCho)(5 mg/kg) alone, arachidonic acid (AA)(5 mg/kg) alone, docosahexaenoic acid (DHA)(5 mg/kg) alone, or 1-palmitoyl-2-linoleil-sn-glycero-3-phosphoserine (PLPhtSer)(5 mg/kg) alone. POPhtCho (5 mg/kg) alone or DLPhtCho (5 mg/kg) plus POPhtCho (5 mg/kg) also significantly shortened the prolonged retention latency for rats intraperitoneally injected with scopolamine, but otherwise no significant effect was obtained with DLPhtCho (5 mg/kg) alone, AA (5 mg/kg) alone, DHA (5 mg/kg) alone, or PLPhtSer (5 mg/kg) alone. Oral co-administration with DLPhtCho (5 mg/kg) and POPhtCho (5 mg/kg) significantly shortened the acquisition latency for rats untreated with scopolamine as compared with the latency for administration with polyethylene glycol (PEG), DLPhtCho alone at doses of 5 and 10 mg/kg, or POPhtCho alone at doses of 5 and 10 mg/kg, while no efficient effect on the retention latency was obtained. To assess the effect of DLPhtCho and POPhtCho on cognitive functions for humans, Mini Mental State Examination (MMSE) test was performed in subjects with cognitive disorders (the average MMSE score, 15). Oral co-intake with DLPhtCho (50 mg) and POPhtCho (45 mg) once after breakfast everyday raised the score to over 20, corresponding to normal cognitive functions, throughout 5 months after intake, and the increase in the score was significantly greater than that for oral intake with DLPhtCho (100 mg/day) alone or POPhtCho (90 mg/kg) alone. Taken together, the results of the present study show that co-intake with DLPhtCho and POPhtCho could enhance learning and memory ability and improve cognitive disorders for both the animals and humans with a promising efficacy.

Topics: Aged; Aged, 80 and over; Animals; Cognition Disorders; Disease Models, Animal; Drug Combinations; Female; Humans; Male; Maze Learning; Memory Disorders; Middle Aged; Nootropic Agents; Phosphatidylcholines; Rats; Rats, Wistar; Scopolamine

Serum paraoxonase (PON1) is a high density lipoprotein (HDL)-associated enzyme involved in organophosphate (OP) degradation and prevention of atherosclerosis. PON1 comprises a potential candidate for in vivo therapeutics, as an anti-atherogenic agent, and for detoxification of pesticides and nerve agents. Because human PON1 exhibits limited stability, engineered, recombinant PON1 (rePON1) variants that were designed for higher reactivity, solubility, stability, and bacterial expression, are candidates for treatment. This work addresses the feasibility of in vivo administration of rePON1, and its HDL complex, as a potentially therapeutic agent dubbed BL-3050.. For stability studies we applied different challenges related to the in vivo disfunctionalization of HDL and PON1 and tested for inactivation of PON1's activity. We applied acute, repetitive administrations of BL-3050 in mice to assess its toxicity and adverse immune responses. The in vivo efficacy of recombinant PON1 and BL-3050 were tested with an animal model of chlorpyrifos-oxon poisoning.. Inactivation studies show significantly improved in vitro lifespan of the engineered rePON1 relative to human PON1. Significant sequence changes relative to human PON1 might hamper the in vivo applicability of BL-3050 due to adverse immune responses. However, we observed no toxic effects in mice subjected to repetitive administration of BL-3050, suggesting that BL-3050 could be safely used. To further evaluate the activity of BL-3050 in vivo, we applied an animal model that mimics human organophosphate poisoning. In these studies, a significant advantages of rePON1 and BL-3050 (>87.5% survival versus <37.5% in the control groups) was observed. Furthermore, BL-3050 and rePON1 were superior to the conventional treatment of atropine-2-PAM as a prophylactic treatment for OP poisoning.. In vitro and in vivo data described here demonstrate the potential advantages of rePON1 and BL-3050 for treatment of OP toxicity and chronic cardiovascular diseases like atherosclerosis. The in vivo data also suggest that rePON1 and BL-3050 are stable and safe, and could be used for acute, and possibly repeated treatments, with no adverse effects.

Topics: Animals; Aryldialkylphosphatase; Chlorpyrifos; Disease Models, Animal; Enzyme Stability; Female; Glutathione; Humans; Injections, Intravenous; Lipoproteins, HDL; Male; Mice; Mice, Inbred C57BL; Organophosphates; Phosphatidylcholines; Protein Engineering; Recombinant Proteins

Irreversible myocardial injury is a potential consequence of coronary artery revascularization. Reperfusion leads to the production of oxidized products that can damage myocardium. High-density lipoproteins (HDL) are effective at removing oxidized lipids. We hypothesized that a synthetic HDL preparation, comprising recombinant apolipoprotein A-I(Milano) (apoA-I(M)) complexed with 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) (apoA-I(M)/POPC) would protect the heart from reperfusion injury. The ex vivo model consisted of rabbit hearts perfused by the Langendorff method. Hearts were equilibrated with Krebs-Henseleit buffer (10 min), pretreated with either apoA-I(M)/POPC (0.45 mg/mL) or vehicle (10 min), subjected to global ischemia (30 min) and reperfused for 60 min. ApoA-I(M)/POPC (n=7) prevented the left ventricular end-diastolic pressure elevation observed in the vehicle group (n=6) at the end of reperfusion (p<0.05). During reperfusion, coronary artery perfusion pressure increased in the controls (p<0.001), but not with apoA-I(M)/POPC. ApoA-I(M)/POPC reduced the release of creatine kinase at the end of the ischemic period (p<0.001). It also reduced cardiac left ventricle muscle lipid hydroperoxides by 46% (p<0.05). Direct comparison of the antioxidant potential indicated that recombinant apoA-I(M) was much more potent than apoA-I in attenuating low-density lipoprotein oxidation. Electron microscopy showed that apoA-I(M)/POPC prevented mitochondrial granulation, disorganization and sarcomere contraction band formation indicative of reperfusion injury. The apoA-I(M)/POPC complex thus appears to reduce reperfusion injury under global ischemic conditions, and may therefore have therapeutic application in the reduction of myocardial ischemia.

Topics: Animals; Antioxidants; Apolipoprotein A-I; Disease Models, Animal; Lipid Peroxidation; Lipoproteins, HDL; Male; Microscopy, Electron; Myocardial Reperfusion Injury; Phosphatidylcholines; Rabbits; Recombinant Proteins; Ventricular Dysfunction