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Benign prostatic hyperplasia (BPH) is a prevalent urological condition that predominantly affects the geriatric male population, resulting in lower urinary tract symptoms. Saw palmetto is a traditional Chinese medicine for treating BPH. This study aimed to investigate the potential therapeutic mechanisms of saw palmetto in BPH treatment. The active ingredients and potential targets of saw palmetto were obtained through the TCMSP database. BPH-related targets were retrieved from the GeneCards database. PPI, GO, and KEEG analyses were performed to predict the potential therapeutic mechanism. The active ingredient-common target and common target-pathway networks were constructed by Cytoscape software. Molecular docking and cellular experiments were carried out to further validate the potential mechanism. We obtained 13 active components in saw palmetto and 56 common targets in BPH treatment. KEEG analysis showed that the estrogen signaling pathway was the most enriched and exhibited a close association with BPH. PPI analysis, along with ingredient-target and target-pathway network analyses, indicated that stigmasterol was the core ingredient and PGR, NCOA1, and NCOA2 were identified as the hub genes mediating the effects of saw palmetto against BPH. In addition, molecular docking showed that stigmasterol had strong binding to PGR, NCOA1, and NCOA2. Cellular experiments revealed that stigmasterol significantly increased the percentage of BPH-1 cells in the G0/G1 phase and inhibited cell viability and division. Furthermore, it notably reduced the expression of PGR, NCOA1, and NCOA2. Saw palmetto might inhibit cell viability and division by suppressing the expression of PGR, NCOA1, and NCOA2, thereby playing a therapeutic role in treating BPH.
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Introduction: A limitation of hemoglobin-based oxygen carriers (HBOCs) as oxygen therapeutics is unpolymerized hemoglobin, which induces vasoconstriction leading to hypertension. The removal of unpolymerized hemoglobin from polymerized hemoglobin (PolyHb) is complex, expensive, and time-consuming. Methods: Herein, we developed a method to completely polymerize hemoglobin almost without unpolymerized hemoglobin. Hemoglobin was adsorbed on the anion-exchange resin Q Sepharose Fast Flow or DEAE Sepharose Fast Flow, and acetal, a crosslinker prepared from glutaraldehyde and ethylene glycol, was employed to polymerize the hemoglobin. The polymerization conditions, including reaction time, pH, resin type, and molar ratios of glutaraldehyde to ethylene glycol and hemoglobin to acetal, were optimized. The blood pressure and blood gas of mice injected with PolyHb were monitored as well. Results: The optimal polymerization condition of PolyHb was when the molar ratio of glutaraldehyde to ethylene glycol was 1:20, and the molar ratio of 10 mg/mL hemoglobin adsorbed on anion-exchange resin to glutaraldehyde was 1:300 for 60 min. Under optimized reactive conditions, hemoglobin was almost completely polymerized, with <1% hemoglobin remaining unpolymerized, and the molecular weight of PolyHb was more centrally distributed. Furthermore, hypertension was not induced in mice by PolyHb, and there were also no pathological changes observed in arterial oxygen, blood gas, electrolytes, and some metabolic indicators. Conclusion: The findings of this study indicate that the use of solid-phase polymerization and acetal is a highly effective and innovative approach to HBOCs, resulting in the almost completely polymerized hemoglobin. These results offer promising implications for the development of new methods for preparing HBOCs.
Asunto(s)
Acetales , Oxígeno , Animales , Ratones , Oxígeno/metabolismo , Glutaral/química , Polimerizacion , Sefarosa , Hemoglobinas/metabolismo , Glicoles de Etileno , AnionesRESUMEN
Excessive reactive oxygen species (ROS), a highly reactive substance that contains oxygen, induced by ultraviolet A (UVA) cause oxidative damage to skin. We confirmed that hemin can catalyze the reaction of tyrosine (Tyr) and hydrogen peroxide (H2O2). Catalysis was found to effectively reduce or eliminate oxidative damage to cells induced by H2O2 or UVA. The scavenging effects of hemin for other free-radical ROS were also evaluated through pyrogallol autoxidation, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·)-scavenging assays, and phenanthroline-Fe2+ assays. The results show that a mixture of hemin and tyrosine exhibits strong scavenging activities for H2O2, superoxide anion (O2-·), DPPH·, and the hydroxyl radical (·OH). Furthermore, the inhibition of oxidative damage to human skin keratinocyte (HaCaT) cells induced by H2O2 or UVA was evaluated. The results show that catalysis can significantly reduce the ratio of cell apoptosis and death and inhibit the release of lactate dehydrogenase (LDH), as well as accumulation of malondialdehyde (MDA). Furthermore, the resistance to apoptosis was found to be enhanced. These results show that the mixture of hemin and tyrosine has a significantly protective effect against oxidative damage to HaCaT cells caused by UVA, suggesting it as a protective agent for combating UVA damage.
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Oxidative damage caused by the ferryl hemoglobin is one of the major clinical adverse reactions of hemoglobin-based oxygen carriers (HBOCs), while the production of reactive oxygen species in a pathological state can oxidize hemoglobin (HbFe2+ ) to ferryl Hb, which can then enter the pseudoperoxidase cycle, making hemoglobin highly toxic. In this study, we found that ferrous hemoglobin and polymerized porcine hemoglobin (one of the HBOCs) have the peroxidase activity different from the pseudoperoxidase activity of ferric hemoglobin. Ferrous hemoglobin can catalyze the reaction of tyrosine (Tyr) with hydrogen peroxide. In addition, the results also indicated that ferrous hemoglobin and pPolyHb have a strong inhibitory effect on the pseudoperoxidase activity of ferric hemoglobin. Therefore, hydrogen peroxide was consumed in a large amount, which greatly prevented hemoglobin from becoming oxidized and entering the pseudoperoxidase cycle, thus inhibiting ferryl Hb toxicity. We further cultured human umbilical vein endothelial cells and monitored cell morphology, viability, cell cycle, apoptosis, lactate dehydrogenase (LDH) release, and malondialdehydes (MDAs) formation when incubated with H2 O2 , Tyr, and HbFe2+ . HbFe2+ and pPolyHb reduced cell cycle arrest, apoptosis, LDH release, and MDA formation. These results showed that reducing oxidative damage induced by H2 O2 and converted hemoglobin from a molecule that is toxic to one that inhibits oxidative damage, suggesting a new strategy for development of a safer HBOCs.
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Sustitutos Sanguíneos/química , Sustitutos Sanguíneos/farmacología , Hemoglobinas/farmacología , Peróxido de Hidrógeno/antagonistas & inhibidores , Animales , Apoptosis/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Hemoglobinas/química , Células Endoteliales de la Vena Umbilical Humana , Humanos , Peróxido de Hidrógeno/metabolismo , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Peroxidasas/química , PorcinosRESUMEN
The factors affecting hemoglobin (Hb) oxidation in its short- and long-term storage were investigated. Porcine Hb with different levels of oxygen saturation was stored at 25°C or 4°C for 40 days. The methemoglobin (metHb) content increased rapidly when Hb was at half saturation with oxygen at 4°C, increased gradually with the increase in Hb oxygenation or deoxygenation, and was almost unchanged when Hb was highly oxygenated or deoxygenated. The Hb oxidation was more intense when stored at 25°C. This indicated that highly oxygenated or deoxygenated Hb can be stored short term at 4°C. The accelerated test showed that metHb content was almost unchanged when Hb was highly deoxygenated and stored at 37°C for 7 days; however, the metHb content rapidly increased when Hb was highly oxygenated and stored at 37°C, which indicated that only highly deoxygenated Hb is suitable for long-term storage. Then, the long-term oxidation stability of highly deoxygenated Hb and polymerized hemoglobin was verified; the metHb content of both did not show significant changes at 4°C for 18 months. Moreover, the results in this work indicated that a temperature increasing from 20 to 40°C clearly increased the partial oxygen pressure (P50), which represents decreased oxygen affinity and is beneficial to deoxygenation. Furthermore, P50 was increased when the pH decreased from 9 to 7. Therefore, we concluded that the appropriate high temperature and neutral condition in vitro are conducive to reducing the oxygen affinity of Hb to achieve efficient deoxygenation, which could promote the development of hemoglobin-based oxygen carriers.
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Hemoglobinas/química , Almacenaje de Medicamentos , Oxidación-ReducciónRESUMEN
A rapid and simple high-performance liquid chromatography-ultraviolet method was developed for the separation and quantification of 15 sulfonamides (SAs) in foods of animal origin without the need of clean-up procedure. A mixture of acetonitrile-formic acid-ammonium acetate-water was used as the mobile phase to separate 15 SAs on a C18 column with gradient. The selected SAs were separated completely from the matrix mixture based on different retention behaviors at different concentration of acetonitrile. The effects of the additive of formic acid and ammonium acetate in mobile phases on the separation of SAs were also investigated. The additive can greatly improve the resolution between SAs and impurities, so that the SAs can be quantified directly under the optimized chromatographic condition the sample preparation which does not need extra sample clean-up procedure. Complete baseline separation of 15 SAs was achieved in <40 min, the linear range is 0.01-130 µg/mL with a correlation coefficient R2-value > 0.999. Excellent method reproducibility was found by intra- and inter-day precisions with the relative standard deviation <9.5%. The detection limit was <11.0 ng/mL and it can be used for routine screening of the SA residues in foods of animal origin.
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Cromatografía Líquida de Alta Presión/métodos , Residuos de Medicamentos/análisis , Carne/análisis , Sulfonamidas/análisis , Animales , Bovinos , Límite de Detección , Modelos Lineales , Leche/química , Reproducibilidad de los Resultados , Espectrofotometría Ultravioleta , PorcinosRESUMEN
Glutaraldehyde (GA), used medically as a disinfectant and as a crosslinker for haemoglobin (Hb)-based oxygen carriers (HBOCs), was investigated for its ability to inactivate viruses during the preparation of these artificial blood substitutes. Porcine parvovirus (PPV; a non-enveloped DNA virus) and porcine pseudorabies virus (PRV; an enveloped DNA virus) were used as the virus indicators. Upon treatment with 0.1 mM GA, the titer of PRV decreased from 9.62 log10 to 2.62 log10 within 0.5 h, whereas that of PPV decreased from 7.00 log10 to 2.30 log10 in 5 h. Following treatment with 1.0 mM GA, the titer of PRV decreased from 11.00 log10 to 1.97 log10 within 0.5 h, whereas that of PPV decreased from 7.50 log10 to 3.43 log10 in 4.5 h. During the polymerization of Hb with GA, the GA concentration decreased to 1.0 and 0.1 mM within 30 and 50 min, respectively, at a GA:Hb molar ratio of 10:1, whereas at a GA:Hb molar ratio of 30:1, GA decreased to those same concentrations in 1.5 and 2.5 h, respectively. This rapid decrease in GA concentration during its polymerization with Hb indicates that GA must be added into the Hb solution in a short time in order to get as high a initial concentration as possible. In this study, the GA can only inactivate PRV effectively, given that a longer time (4.5 h) was required for it to inactivate the PPV titer. This study therefore demonstrates that GA inactivates the enveloped DNA virus only during the preparation of HBOCs.
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Glutaral/farmacología , Hemoglobinas/metabolismo , Herpesvirus Suido 1/efectos de los fármacos , Oxígeno/metabolismo , Parvovirus Porcino/efectos de los fármacos , Inactivación de Virus/efectos de los fármacos , Animales , Línea Celular , Herpesvirus Suido 1/fisiología , Parvovirus Porcino/fisiología , Polimerizacion , SeguridadRESUMEN
A Tempol compound with an amine group (4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl, NH2-Tempol) was cross-linked to hemoglobin in a one-step polymerization reaction to produce a novel hemoglobin-based oxygen carrier (HBOC) designated PolyHb-Tempol. The reaction parameters, including the reaction time, pH, temperature, and ratio of reactants, were optimized, and the physiochemical properties of the resulting product were characterized. PolyHb-Tempol didn't show any toxicity towards endothelial cells. Furthermore, from observations of cell morphology and viability, PolyHb-Tempol showed a significant ability to inhibit or eliminate oxidative stress induced by superoxide free radicals. These results suggest that PolyHb-Tempol may potentially be suitable as an HBOC.
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Óxidos N-Cíclicos/química , Células Endoteliales/efectos de los fármacos , Glutaral/química , Hemoglobinas/farmacología , Estrés Oxidativo/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Superóxidos/farmacología , Supervivencia Celular/efectos de los fármacos , Células Endoteliales/citología , Células Endoteliales/metabolismo , Hemoglobinas/química , Concentración de Iones de Hidrógeno , Cinética , Polimerizacion , Marcadores de Spin , TemperaturaRESUMEN
The effects of polymerized porcine hemoglobin (pPolyHb) on hemodynamic stability and maintenance of mesenteric microvascular function were explored in a rat model of hemorrhagic shock (HS). Following controlled hemorrhage, rats were infused with equal volumes of either pPolyHb, hetastarch (HES), or red blood cell (RBC). The results showed that pPolyHb was superior to HES and RBC in restoring hemodynamic stability and reversing anaerobic metabolism. We observed a reduction in the diameter of mesenteric microvasculature after HS. Resuscitation with pPolyHb and RBC was able to restore the diameters of the venules and arterioles, whereas HES failed to restore the diameters during the observation period.
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Hemodinámica/efectos de los fármacos , Hemoglobinas/farmacología , Arterias Mesentéricas/efectos de los fármacos , Arterias Mesentéricas/fisiopatología , Microcirculación/efectos de los fármacos , Choque Hemorrágico/tratamiento farmacológico , Choque Hemorrágico/fisiopatología , Acidosis/complicaciones , Animales , Biomarcadores/metabolismo , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Frecuencia Cardíaca/efectos de los fármacos , Hemoglobinas/uso terapéutico , Masculino , Ratas , Choque Hemorrágico/complicaciones , Choque Hemorrágico/metabolismo , PorcinosRESUMEN
The objective of this study was to evaluate the effects of polymerized porcine hemoglobin (pPolyHb) on hemodynamic stability and oxygen delivery in a rat model of hemorrhagic shock. Rats underwent controlled hemorrhage, resulting in the loss of 65% of their blood volume in 90 minutes. The results showed that pPolyHb was superior to hetastarch and saline, and similar to whole blood, in restoring hemodynamic stability and reversing anaerobic metabolism caused by hemorrhage. Furthermore, pPolyHb improved oxygen supply and increased blood oxygen content. These data suggest that pPolyHb can be effective in improving tissue perfusion under conditions of severe hemorrhagic shock.
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Sustitutos Sanguíneos/farmacología , Hemodinámica/efectos de los fármacos , Hemoglobinas/farmacología , Oxígeno/sangre , Resucitación/métodos , Choque Hemorrágico , Animales , Modelos Animales de Enfermedad , Masculino , Ratas , Ratas Sprague-Dawley , Choque Hemorrágico/sangre , Choque Hemorrágico/tratamiento farmacológico , PorcinosRESUMEN
The objective of this study was to investigate the recovery ability of polymerized porcine hemoglobin (pPolyHb) in a rat model of acute anemia caused by normovolemic hemodilution (ANH). After the ANH procedure, rats were infused with either pPolyHb or red blood cells. The results showed pPolyHb could carry a sufficient amount of oxygen to the tissues to maintain normal aerobic metabolism and hemodynamic stability, without any significant toxic effects on renal and liver function according to pathological, and biochemical analysis. The data suggest pPolyHb may be a good candidate for the treatment of acute anemia in future clinical trials.
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Anemia/tratamiento farmacológico , Sustitutos Sanguíneos/farmacología , Hemoglobinas/farmacología , Porcinos , Enfermedad Aguda , Anaerobiosis/efectos de los fármacos , Anemia/sangre , Anemia/metabolismo , Anemia/fisiopatología , Animales , Presión Sanguínea/efectos de los fármacos , Sustitutos Sanguíneos/uso terapéutico , Modelos Animales de Enfermedad , Frecuencia Cardíaca/efectos de los fármacos , Hemoglobinas/uso terapéutico , Concentración de Iones de Hidrógeno , Masculino , Oxígeno/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
A rapid and simple high-performance liquid chromatography-UV method was developed for the separation and quantification of salbutamol, ractopamine, and clenbuterol in pork. A mixture of acetonitrile-formic acid-ammonium acetate was used as the mobile phase to separate three ß-agonists on a C18 column with gradient. The effects of the addition of formic acid and ammonium acetate to mobile phases on the separation of ß-agonists were investigated. These additives can greatly improve the resolution and sensitivity. Under the optimized chromatographic condition, this separation does not need extra sample preparation. Complete baseline separation of three ß-agonists was achieved in < 20 minutes; the linear range is 0.2-50 µg/L with a correlation coefficient R2 value of > 0.99. Excellent method reproducibility was found by intra- and interday precisions with a relative standard deviation of < 3%. The detection limit (S/N = 3) was found to be <0.05 µg/L; this method can be used for routine screening of the ß-agonist residues in foods of animal origin before being identified by confirmatory methods.
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Carne Roja , Albuterol , Animales , Cromatografía Líquida de Alta Presión , Clenbuterol , Análisis de los Alimentos , Fenetilaminas , Reproducibilidad de los Resultados , PorcinosRESUMEN
Polymerized porcine hemoglobin (pPolyHb) exhibits a protective effect on ischemia/reperfusion of organ grafts. A series of experiments were performed to explore the underlying cytoprotective mechanisms of pPolyHb pretreatment on H2O2-induced cell death and apoptosis. The results showed that the pretreatment augmented heme oxygenase-1 (HO-1) expression, and at the same time, decreased the phosphorylation of JNK/p38 mitogen-activated protein kinase (MAPK) and intracellular ROS generation in H2O2-treated HUVECs. Moreover, the inhibition of HO-1 expression by tin porphyrin (SnPP) abolished the protective effects of pPolyHb, which suggested that the cytoprotective effect of pPolyHb involves upregulating HO-1 and subsequently decreasing the phosphorylation of the JNK and p38 MAPK and ROS generation.
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Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Hemo-Oxigenasa 1/biosíntesis , Hemoglobinas/farmacología , Células Endoteliales de la Vena Umbilical Humana/enzimología , Peróxido de Hidrógeno/farmacología , MAP Quinasa Quinasa 4/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Endotelio Vascular/enzimología , Endotelio Vascular/lesiones , HumanosRESUMEN
Small bowel transplantation (SBTx) has become a standard clinical treatment for short bowel syndrome or irreversible intestinal function failure. Optimum preservation of the organ is essential for the success of transplantation. In this study, pPolyHb was used as an additive to hypertonic citrate adenine solution (HCA) to provide oxygen for rat small bowel transplant. Rat small bowels were preserved in HCA, HCA with pPolyHb, and University of Wisconsin solution (UW) for 12, 24, and 36 h, respectively. The results suggested that the preservation effect of HCA with pPolyHb was comparable with the UW solution, and more effective than the HCA solution.
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Hemoglobinas/química , Intestino Delgado , Preservación Biológica/métodos , Multimerización de Proteína , Adenina/farmacología , Adenosina Trifosfato/metabolismo , Animales , Análisis de los Gases de la Sangre , Intestino Delgado/citología , Intestino Delgado/efectos de los fármacos , Intestino Delgado/metabolismo , Masculino , Ratas , PorcinosRESUMEN
Hemoglobin-based oxygen carriers (HBOCs), with their capacity for delivering oxygen, could potentially function as red blood cell substitutes or primary resuscitation solutions. However, there has been some concern regarding redox-related safety issues of HBOCs. The present study describes a novel function of polymerized porcine hemoglobin (pPolyHb) in protecting a human umbilical vein endothelial cell line from H2O2-induced cytotoxicity. Through the examination of H2O2 consumption and ferrylhemoglobin formation, we found that pPolyHb exhibits antioxidant activity, suggesting that pPolyHb may protect cells from free radical-induced cell damage. Additionally, we investigated the effect of pPolyHb on H2O2-induced cell cytotoxicity, and found that pPolyHb significantly inhibits H2O2-mediated endothelial cell damage as well as apoptosis. Thus, pPolyHb may be developed as a new HBOC in the future.
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Sustitutos Sanguíneos/química , Sustitutos Sanguíneos/farmacología , Citotoxinas/antagonistas & inhibidores , Células Endoteliales/efectos de los fármacos , Peróxido de Hidrógeno/antagonistas & inhibidores , Animales , Apoptosis/efectos de los fármacos , Citotoxinas/metabolismo , Células Endoteliales/citología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Peróxido de Hidrógeno/metabolismo , Polimerizacion , PorcinosRESUMEN
Composite nanoparticles have proved to be promising in a wide range of biotechnological applications. In this paper, we report on a facile method to synthesize novel Fe(3)O(4)/Au/Fe(3)O(4) nanoparticles (nanoflowers) that integrate hybrid components and surface types. We demonstrate that relative to conventional nanoparticles with core/shell configuration, such nanoflowers not only retain their surface plasmon property but also allow for 170% increase in the saturation magnetization and 23% increase in the conjugation efficiency due to the synergistic co-operation between the hierarchical structures. Moreover, we demonstrate that the magnetic properties of such composite nanoparticles can be tuned by controlling the size of additional petals (Fe(3)O(4) phase). These novel building blocks could open up novel and exciting vistas in nanomedicine for broad applications such as biosensing, cancer diagnostics and therapeutics, targeted delivery, and imaging.
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Nanopartículas de Magnetita/química , Animales , Inmunoconjugados/química , Inmunoglobulina G , Magnetismo , Nanopartículas de Magnetita/ultraestructura , Microscopía Electrónica de Transmisión , Nanocompuestos/química , Nanocompuestos/ultraestructura , Nanotecnología , Espectrofotometría , Difracción de Rayos XRESUMEN
Polymerized Porcine Hemoglobin (pPolyHb), a hemoglobin-based oxygen carrier (HBOC), was developed as a potential red blood substitute for clinical applications. Assessment of its effects on the immune system is an important component of the overall safety evaluation of HBOC. For this purpose, we assessed three inflammation indicators, including complement C3a, IL-6, and TNF-? in cultured cells and in a rat model when pPolyHb was incubated or administrated with the cells/animals. Our results suggested that the levels of these three indicators were not statistically changed upon pPolyHb stimulation, indicating that pPolyHb is not immunotoxic to cells and animals in this aspect.
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Sustitutos Sanguíneos/efectos adversos , Sustitutos Sanguíneos/química , Eritrocitos , Hemoglobinas/efectos adversos , Hemoglobinas/química , Inmunidad Innata/efectos de los fármacos , Porcinos , Animales , Biopolímeros/inmunología , Complemento C3a/metabolismo , Hemoglobinas/inmunología , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/metabolismo , Interleucina-6/metabolismo , Masculino , Multimerización de Proteína , Estructura Cuaternaria de Proteína , Ratas , Ratas Sprague-Dawley , Choque Hemorrágico/inducido químicamente , Choque Hemorrágico/inmunología , Choque Hemorrágico/metabolismo , Porcinos/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The objective of the present study is to evaluate the pharmacodynamic properties of polymerized porcine hemoglobin (pPolyHb) in an exchange transfusion model. Each of two groups of rats received a volume of pPolyHb or hetastarch that equalled 120-140% of estimated total blood volume (70 ml/kg) exchange transfusion. The results showed pPolyHb retained hemodynamic stability and exhibited superior volume expansion capability. Furthermore, pPolyHb effectively reverse anaerobic metabolism caused by a large amount of volume exchange. In comparison with hetastarch, pPolyHb increased blood oxygen content and tissue oxygenation. All these properties contribute to a higher effectiveness in sustaining the lives of rats in pPolyHb group.
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Recambio Total de Sangre , Hemoglobinas/farmacocinética , Consumo de Oxígeno , Oxígeno , Polimerizacion , Anaerobiosis/efectos de los fármacos , Animales , Glutaral/química , Hemodinámica/efectos de los fármacos , Hemoglobinas/administración & dosificación , Hemoglobinas/química , Derivados de Hidroxietil Almidón/administración & dosificación , Masculino , Modelos Animales , Oxígeno/sangre , Consumo de Oxígeno/efectos de los fármacos , Ratas/inmunología , Ratas Sprague-Dawley , PorcinosRESUMEN
This study introduced a liquid-liquid extraction method designed for complete recovery of phospholipids from protein-rich samples, such as hemoglobin (Hb) solutions. In order to minimize protein denaturation and maximize lipid extraction from protein-rich samples, isopropanol was used as the major organic extraction solvent. In a wide range of the volume ratio of isopropanol to Hb solution at low temperatures, such extraction system resulted in limited protein precipitation and did not cause heme pigment contamination. The efficiency of phospholipid extraction from Hb solutions was pH-dependent, with the lowest extraction yield around neural pH and the highest extraction yield around pH 5.0. At pH 5.0, salt with concentrations up to 400 mmol/L in the samples did not increase extraction efficiency. Compared to other available methods, this method is simpler, needs significantly less organic solvent and, most importantly, consumed much less Hb, which is very expensive to make for the purpose of production of therapeutic products, hemoglobin-based oxygen carriers (HBOCs). This method is suitable not only for Hb solutions but also for other protein-rich biological samples, such as erythrocytes, plasma and liver.