RESUMEN
The regeneration of absorbed alveolar bone and reconstruction of periodontal support tissue are huge challenges in the clinical treatment of periodontitis due to the limited regenerative capacity of alveolar bone. It is essential to regulate inflammatory reaction and periodontal cell differentiation. Based on the anti-inflammatory effect of baker's yeast ß-glucan (BYG) with biosafety by targeting macrophages, the BYG-based nanoparticles loading methotrexate (cBPM) were fabricated from polyethylene glycol-grafted BYG through chemical crosslinking for treatment of periodontitis. In our findings, cBPM promoted osteogenesis of human dental pulp stem cells (hDPSCs) under inflammatory microenvironment, characterized by the enhanced expression of osteogenesis-related Runx2 and activation of mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/Erk) pathway in vitro. Animal experiments further demonstrate that cBPM effectively promoted periodontal bone regeneration and achieved in a better effect of recovery indicated by 19.2 % increase in tissue volume, 7.1 % decrease in trabecular separation, and a significant increase in percent bone volume and trabecular thickness, compared with the model group. Additionally, cBPM inhibited inflammation and repaired alveolar bone by transforming macrophage phenotype from inflammatory M1 to anti-inflammatory M2. This work provides an alternative strategy for the clinical treatment of periodontitis through BYG-based delivery nanoplatform of anti-inflammatory drugs.
Asunto(s)
Regeneración Ósea , Pulpa Dental , Metotrexato , Nanopartículas , Osteogénesis , beta-Glucanos , Humanos , Osteogénesis/efectos de los fármacos , Nanopartículas/química , Regeneración Ósea/efectos de los fármacos , beta-Glucanos/farmacología , beta-Glucanos/química , Pulpa Dental/efectos de los fármacos , Pulpa Dental/citología , Animales , Metotrexato/farmacología , Metotrexato/química , Células Madre/efectos de los fármacos , Periodontitis/tratamiento farmacológico , Periodontitis/patología , Masculino , Ratones , Inflamación/tratamiento farmacológico , Portadores de Fármacos/química , Células Cultivadas , Diferenciación Celular/efectos de los fármacosRESUMEN
This study demonstrates that the purified ß-glucan (LNT) with a triple helix and relatively narrow molecular weight distribution, extracted and purified from artificially cultured Lentinus edodes, showed a significant cervical cancer inhibition with little cytotoxicity against normal cells in vitro and in vivo. From the in vitro data, the potential mechanism of anti-cervical cancer was preliminarily revealed as follows: LNT was firstly recognized by the human cervical cancer cell line of Hela and induced cell proliferation inhibition through p21 and apoptosis via a mitochondrion-dependent pathway by targeting the tumor suppressor of p53, indicated by an increase in reactive oxygen species (ROS) generation and a loss of mitochondrial membrane potential (Δψm), in a significant dosage-dependent manner. Meanwhile, LNT repressed tumor growth with an inhibition ratio of 61.2 % and induced tumor cell apoptosis through endogenous MDM2/p53/Bax/mitochondrion signal pathway by up-regulating the expression of p53, Bax, Cyt. c, caspase 9, and caspase 3, as well as down-regulating Bcl-2, MDM2, and PARP1 levels in Hela cells-transplanted BALB/c nude mice. This study provides a scientific basis for the clinical treatment of cervical cancer with LNT as a potential drug candidate characterized by the triple helix and specified molecular weight with a relatively narrow distribution.
RESUMEN
The linear ß-(1, 3)-glucans from yeast (BYGs) with good biocompatibility and targetability to macrophages were used for fabricating BYG-based nanoparticles to deliver methotrexate with systemic toxicity for treatment of rheumatoid arthritis. Methoxy poly (ethylene glycol) (mPEG) was successfully grafted onto BYGs chains, followed by chemical crosslinking to get the crosslinked copolymer (cBP) with amphiphilicity, which could self-assemble into spherical nanoparticles (ca.52.9 nm in diameter). The methotrexate-loaded cBP nanoparticles (cBPM) with the drug loading efficiency of 23.7% was proved to linearly release methotrexate due to reduction of disulfide bonds by glutathione. Cell experiments demonstrate that cBP nanoparticles were effectively internalized into macrophages due to the targetability. Animal experiments show that cBPM were highly targeted to the inflamed tissue, leading to macrophage transformation from M1 to M2 type and reduction of pro-inflammatory factors. This work provides an alternative safe strategy for the clinical treatment of rheumatoid arthritis with ß-glucan nanoparticles as carrier.
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Artritis Reumatoide , Nanopartículas , beta-Glucanos , Animales , Artritis Reumatoide/tratamiento farmacológico , Portadores de Fármacos/química , Metotrexato/farmacología , Metotrexato/uso terapéutico , Nanopartículas/química , Saccharomyces cerevisiaeRESUMEN
Collagen and polysaccharide materials have great advantages for burn wound healing and skin regeneration. The purpose of this study was to develop a promising burn dressing. Mixing hyaluronic acid (HA), carboxylated chitosan (CCS) and human-like collagen (HLC) to simulate extracellular matrix (ECM), and glutamine transaminase (TG) was used as a crosslinker. The mechanical properties and pore size of the hydrogel were optimized by response surface methodology. The results showed that the tensile elastic modulus of the hydrogel was 480.43 ± 15.82 kPa, the tensile strain was 55.23 ± 2.43 %, and the pore size was 90.43 ± 5.57 µm. This study constructed a skin burn model and demonstrated that the hydrogel dressing could effectively prevent bacteria infection and confirmed that the hydrogel dressing was more beneficial for promoting burn wound healing than a commercial film (DUO DERM). Therefore, the HLC/HA/CCS hydrogel can be considered a promising burn wound dressing.
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Materiales Biocompatibles/farmacología , Quemaduras/tratamiento farmacológico , Colágeno/farmacología , Hidrogeles/farmacología , Polisacáridos/farmacología , Cicatrización de Heridas/efectos de los fármacos , Animales , Vendajes , Materiales Biocompatibles/síntesis química , Materiales Biocompatibles/química , Colágeno/química , Hidrogeles/química , Polisacáridos/química , ConejosRESUMEN
Jumbo squid (Dosidicus gigas) muscle is rather hard and tough, which directly affects consumer acceptance. In this study, the tenderization effect of bromelain and papain on squid muscle during enzymolysis is examined and compared with an untreated control and water-treated sample. Squid mantle were incubated with different solutions (water, bromelain, and papain solution) for 40 min in a 30 °C water bath. Then, the mantle samples were subjected to water holding capacity (WHC) analysis, texture evaluation, biochemical determination, and histological observations. The results revealed that bromelain and papain disadvantageously decrease the water holding capacity when compared to the control and water-treated samples. Furthermore, following tenderization with bromelain or papain, muscle hardness, shear force, myofibrillar protein content, and Ca2+ ATPase activity were all significantly decreased. Additionally, some essential amino acids were released following tenderization. When examining the myofibrillar fragmentation index (MFI), bromelain and papain were shown to cause high levels of hydrolysis in myofibrillar and sarcoplasmic proteins. Moreover, microstructural imaging indicated that the tenderization treatments disrupted myofibrils and generated a larger number of small fragments in the muscle tissues, subsequently decreasing microstructure stability and integrity. SDS-PAGE analysis confirmed that bromelain and papain have a high proteolytic activity, with some small peptides and/or short fragments detected post-tenderization. The results presented herein demonstrated that bromelain and papain improved squid muscle tenderness and can be utilized to ensure a more desirable squid product.
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Bromelaínas/química , Carne/análisis , Papaína/química , Animales , Decapodiformes , Manipulación de Alimentos , Proteínas Musculares/química , Proteolisis/efectos de los fármacos , Resistencia al Corte , AguaRESUMEN
Diabetic nephropathy (DN), a kind of microvascular complication, is a primary cause of end-stage renal disease worldwide. However, therapeutic drugs for DN treatment are still in lack. The glomerular endothelium is essential to maintain selective permeability of glomerular filtration barrier and glomerular vasculature function. Growing evidences show that endothelial dysfunction or injury is the initial stage of vascular damage in DN, which can be induced by hyperglycemia, lipotoxicity, and inflammation. Therefore, to improve the function of vascular endothelium in kidney is a key point for treatment of DN. As a plant isoflavone, tectorigenin (TEC) has attracted considerable attention due to its anti-proliferative and anti-inflammatory functions. However, whether TEC could inhibit the DN development remains unknown. In this study, we examined the effects of TEC on DN development in db/db mice, a type of genetic defect diabetic mice that can spontaneously develop into severe renal dysfunction. Intriguingly, TEC treatment restored diabetes-induced glucose and lipid metabolic disorder; and improved the deterioration of renal function, particularly the renal endothelium function in db/db mice. Additionally, TEC inhibited the renal inflammation via reducing macrophages infiltration and M1 polarization. Moreover, TEC inhibited lipopolysaccharide (LPS)-induced endothelial injury and M1 polarization in vitro. Mechanistically, TEC partially restored the reduction in expression of adiponectin receptor 1/2 (AdipoR1/2), pi-LKB1, pi-AMPKα, and PPARα in vitro and in vivo. Noteworthy, these beneficial pharmacological activities mediated by TEC were significantly attenuated after AdipoR1/2 knockdown by siRNA, indicating that AdipoR1/2 plays a critical role in protection against DN. Collectively, these results suggested that TEC have a potently effect for retarding type 2 diabetes-associated DN.
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Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Nefropatías Diabéticas/tratamiento farmacológico , Isoflavonas/uso terapéutico , Receptores de Adiponectina/metabolismo , Animales , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Nefropatías Diabéticas/metabolismo , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Técnicas de Silenciamiento del Gen , Humanos , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/metabolismo , Lipopolisacáridos/farmacología , Ratones Endogámicos , Receptores de Adiponectina/genética , Transducción de Señal , Células THP-1RESUMEN
Temperature fluctuation is a common problem in the frozen storage of shrimp products. This study investigated the influence of carrageenan oligosaccharide (CO) and xylooligosaccharide (XO) on the growth and recrystallization of ice crystals in frozen peeled shrimp exposed to temperature fluctuations. Shrimp soaked with water and 3.0% (w/v) Na4P2O7 solution were designated as the negative and positive controls, respectively. Our data revealed that both CO- and XO-soaked shrimp had significant improvements in thawing and cooking loss, myofibrillar protein content, Ca2+-ATPase activity, and textural variables when exposed to temperature fluctuations compared to control samples. Microstructural imaging indicated that soaking the shrimp in CO and XO slowed the progression of damage caused to tissue myofibrils by large ice crystals, as well as inhibited the growth and recrystallization of ice crystals in muscle tissues. SDS-PAGE analysis confirmed that treatment with the oligosaccharides exhibited marked effects on the stability of muscle proteins and inhibited the degradation of muscle proteins affected by the temperature fluctuations. Based on these data, we hypothesize that the incorporated CO and XO may bind to muscle proteins and capture water molecules in the myofibrillar network through hydrogen bonding, thereby suppressing the myofibrillar denaturation and tissue structure destruction induced by the growth and recrystallization of ice crystals.
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Carragenina/química , Glucuronatos/química , Oligosacáridos/química , Penaeidae/química , Animales , Congelación , Enlace de Hidrógeno , Hielo/análisis , Temperatura , Agua/químicaRESUMEN
Formation of thrombosis is associated with activation of platelets and endothelial cells. The effect of LongShengZhi Capsule (LSZ), a traditional Chinese medicine used for treatment of vascular diseases, on thrombosis was investigated in this study. BALB/c mice were induced thrombosis by injection of carrageenan while receiving pre or simultaneous LSZ treatment. We also compared the therapeutic effects of LSZ and clopidogrel on formed thrombi. LSZ inhibited carrageenan-induced thrombi in mouse tissue vessels. In addition, LSZ but not clopidogrel reduced formed thrombi with a short time window. The reduction of thrombi by LSZ was associated with reduced serum P-selectin, reduced expression of TNF-α and P-selectin and activated matrix metalloproteinase 2 expression in tissues. In vitro, LSZ decreased thrombin-induced human platelet clot retraction which was associated with inactivation of AKT and ERK1/2. LSZ also reduced adhesion of platelets or THP-1 monocytes to human umbilical vein endothelial cells (HUVECs) induced by oxidized low-density lipoprotein or lipopolysaccharide. The anti-adherent actions of LSZ was attributed to reduction of oxidative stress, expression of platelet receptors (P2Y12, PAR4 and CD36) and AKT activity in platelets. LSZ also reduced adhesion molecules or tissue factor but activated tissue factor pathway inhibitor expression in HUVECs. Taken together, our study demonstrates the antithrombotic properties of LSZ by reducing activation of platelets and endothelial cells, and suggests its potential application in clinics.
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Medicamentos Herbarios Chinos/uso terapéutico , Células Endoteliales/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Activación Plaquetaria/efectos de los fármacos , Trombosis/tratamiento farmacológico , Animales , Plaquetas/efectos de los fármacos , Plaquetas/patología , Carragenina , Células Endoteliales/patología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Masculino , Ratones Endogámicos BALB C , Trombosis/inducido químicamente , Trombosis/patologíaRESUMEN
Cryoprotective saccharides are widely accepted antifreeze additives that reduce thawing loss, maintain texture, and retard protein denaturation in frozen seafood. In this study, the inhibition effects of trehalose and alginate oligosaccharides on ice growth were investigated and compared with sodium pyrophosphate (Na4P2O7) treatment in peeled shrimp during frozen storage, especially the interactions between saccharide molecules and ice crystals. The microstructural results demonstrated that the pre-soaking of trehalose and alginate oligosaccharides before freezing exhibited marked effects on stability of muscle tissue structures and slowed the damage caused to the myofibrils by large ice crystals. The ice-growth inhibition activities might play an important role in cryoprotective effects of saccharides on frozen muscle tissue. Furthermore, molecular docking and molecular dynamics (MD) simulations proved that saccharides were generally close to the ice interface and embedded in ice layers via hydrogen bonds or hydrophobic or electrostatic interactions. The saccharides-ice complex was partially destroyed, and some dislocation and disaggregation were observed around the saccharides molecules. Thus, the incorporated saccharides suppressed the growth of ice crystals, providing protection from freeze-induced damage. Here, the obtained structural details of the ice crystals interface affected by trehalose and alginate oligosaccharides were well in agreement with the histological (H&E staining) experimental results. These findings help better understand the ice-growth inhibition mechanisms of saccharides in frozen shrimp, and these two saccharides may be potentially used as ice-growth inhibitors in frozen seafood.
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Congelación , Hielo/análisis , Oligosacáridos/química , Trehalosa/química , Alginatos/química , Animales , Difosfatos/química , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Penaeidae/química , Electricidad EstáticaRESUMEN
The objective of the present study was to formulate eprosartan mesylate loaded nano-bilosomes and investigates its potential for controlling streptozotocin induced diabetes nephropathy in Wistar rats. The eprosartan mesylate loaded nano-bilosomes comprising of various ratios of soybean phosphatidylcholine/sodium deoxycholate were prepared by thin film hydration technique. The prepared formulations were evaluated for vesicles size, polydispersity index, zeta potential and entrapment efficiency. Further the optimized formulation was characterized for vesicles morphology, and its efficacy for the management of diabetic nephropathy in Wistar rats. The optimized eprosartan mesylate loaded nano-bilosomes exhibited vesicles size, polydispersity index, zeta potential and entrapment efficiency of 63.88±3.46nm, 0.172±0.026, -30.40±2.75mV and 61.19±0.88% respectively. In vivo activity demonstrated that the prepared eprosartan mesylate loaded nano-bilosomes formulation demonstrated a nephro-protecting outcome as shown by the substantial decrease in serum creatinine, urea, lactate dehydrogenase, total albumin, and malondialdehyde. Additionally, an oral administration of eprosartan mesylate loaded nano-bilosomes decreases the raised expressions of Angiotensin II type 1 receptor, inducible nitric oxide synthase, and transforming growth factor-ß1 in Wistar rats. Further, histopathological examination established the nephro-protective effect of prepared formulation. In conclusion, the research work in the paper suggests that the prepared eprosartan mesylate loaded nano-bilosomes could serve as a practical oral formulation for diabetic nephropathy in future therapy and may offer potential benefits in cases with hypertension and renal disease.
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Diabetes Mellitus Experimental/complicaciones , Nefropatías Diabéticas/prevención & control , Imidazoles/química , Imidazoles/farmacología , Tiofenos/química , Tiofenos/farmacología , Administración Oral , Bloqueadores del Receptor Tipo 1 de Angiotensina II/química , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Animales , Ácidos y Sales Biliares , Portadores de Fármacos , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Liposomas , Masculino , Nanoestructuras , Tamaño de la Partícula , RatasRESUMEN
Intestinal anti-inflammatory activities of exopolysaccharide from S. commune were assessed using dextran sulfate sodium (DSS)-induced colitis in mice model. The changes of molecular weight (MW), atomic force microscope morphology, X-ray diffraction, particle size distribution, and viscosity were recorded after sonication treatment. The results indicated that the triple helical structure of exopolysaccharide was dissociated into single helical structure and random coiled structure by ultrasonication via breaking of inter- and intramolecular hydrogen bonds. The medium (936kDa) and high MW (1437kDa) exopolysaccharide had the mixture of triple helix and single helix conformation, while the low MW (197kDa) exopolysaccharide exhibit random coiled conformation. The intestinal anti-inflammatory activity study showed that oral administration of medium and high MW (1437kDa) exopolysaccharide significantly recovered DSS-induced colitis in inflamed tissues and reduced inflammation induced infiltration of macrophages. These results showed that medium (936kDa) and high MW (1437kDa) exopolysaccharide had intestinal anti-inflammatory activity. The intestinal anti-inflammatory activity of exopolysaccharide was related to helical structure and molecular weight.
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Antiinflamatorios/química , Colitis/tratamiento farmacológico , Intestinos/efectos de los fármacos , Polisacáridos/química , Schizophyllum/química , Animales , Colitis/inducido químicamente , Sulfato de Dextran , Masculino , Ratones , Ratones Endogámicos BALB C , Peso MolecularRESUMEN
Magnesium isoglycyrrhizinate as a hepatoprotective agent possesses immune modulation and anti-inflammation, and treats liver diseases. But its effects on immunological-inflammatory and metabolic profiles for metabolic syndrome with liver injury and underlying potential mechanisms are not fully understood. In this study, magnesium isoglycyrrhizinate alleviated liver inflammation and lipid accumulation in fructose-fed rats with metabolic syndrome. It also suppressed hepatic inflammatory signaling activation by reducing protein levels of phosphorylation of nuclear factor-kappa B p65 (p-NF-κB p65), inhibitor of nuclear factor kappa-B kinase α/ß (p-IKKα/ß) and inhibitor of NF-κB α (p-IκBα) as well as nucleotide-binding domain (NOD)-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC) and Caspase-1 in rats, being consistent with its reduction of interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and IL-6 levels. Furthermore, magnesium isoglycyrrhizinate modulated lipid metabolism-related genes characterized by up-regulating peroxisome proliferator-activated receptor-α (PPAR-α) and carnitine palmitoyl transferase-1 (CPT-1), and down-regulating sensor for fatty acids to control-1 (SREBP-1) and stearoyl-CoA desaturase 1 (SCD-1) in the liver of fructose-fed rats, resulting in the reduction of triglyceride and total cholesterol levels. These effective actions were further confirmed in fructose-exposed BRL-3A and HepG2 cells. The molecular mechanisms underpinning these observations suggest that magnesium isoglycyrrhizinate may inhibit NF-κB/NLRP3 inflammasome activation to reduce immunological-inflammatory response, which in turn may prevent liver lipid metabolic disorder and accumulation under high fructose condition. Thus, blockade of NF-κB/NLRP3 inflammasome activation and lipid metabolism disorder by magnesium isoglycyrrhizinate may be the potential therapeutic approach for improving fructose-induced liver injury with metabolic syndrome in clinic.
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Fructosa/efectos adversos , Inflamasomas/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Hígado/efectos de los fármacos , FN-kappa B/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Saponinas/farmacología , Triterpenos/farmacología , Animales , Regulación hacia Abajo/efectos de los fármacos , Células Hep G2 , Humanos , Hígado/metabolismo , Hígado/patología , Masculino , PPAR alfa/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (Lin Zhi) has been used to treat diabetes in Chinese folk for centuries. Our laboratory previously demonstrated that Ganoderma lucidum polysaccharides (GLPs) had hypoglycemic effects in diabetic mice. Our aim was to identify the main bioactives in GLPs and corresponding mechanism of action. MATERIALS AND METHODS: Four polysaccharide-enriched fraction were isolated from GLPs and the antidiabetic activities were evaluated by type 2 diabetic mice. Fasting serum glucose (FSG), fasting serum insulin (FSI) and epididymal fat/BW ratio were measured at the end of the experiment. In liver, the mRNA levels of hepatic glucose regulatory enzymes were determined by quantitative polymerase chain reaction (qPCR) and the protein levels of phospho-AMP-activated protein kinase (p-AMPK)/AMPK were determined by western blotting test. In epididymal fat tissue, the mRNA and protein levels GLUT4, resistin, fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC1) were determined by qPCR and immuno-histochemistry. The structure of polysaccharide F31 was obtained from GPC, FTIR NMR and GC-MS spectroscopy, RESULTS: F31 significantly decreased FSG (P<0.05), FSI and epididymal fat/BW ratio (P<0.01). In liver, F31 decreased the mRNA levels of hepatic glucose regulatory enzymes, and up-regulated the ratio of phospho-AMP-activated protein kinase (p-AMPK)/AMPK. In epididymal fat tissue, F31 increased the mRNA levels of GLUT4 but decreased fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC1) and resistin. Immuno-histochemistry results revealed F31 increased the protein levels of GLUT4 and decreased resistin. CONCLUSION: Data suggested that the main bioactives in GLPs was F31, which was determined to be a ß-heteropolysaccharide with the weight-average molecular weight of 15.9kDa. The possible action mechanism of F31 may be associated with down-regulation of the hepatic glucose regulated enzyme mRNA levels via AMPK activation, improvement of insulin resistance and decrease of epididymal fat/BW ratio. These results strongly suggest that F31 has antidiabetic potential.
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Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Polisacáridos Fúngicos , Ganoderma , Hipoglucemiantes , Proteínas Quinasas Activadas por AMP/metabolismo , Acetil-CoA Carboxilasa/genética , Acetil-CoA Carboxilasa/metabolismo , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Animales , Glucemia/análisis , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/metabolismo , Regulación hacia Abajo , Ayuno/sangre , Acido Graso Sintasa Tipo I/genética , Acido Graso Sintasa Tipo I/metabolismo , Cuerpos Fructíferos de los Hongos , Polisacáridos Fúngicos/farmacología , Polisacáridos Fúngicos/uso terapéutico , Transportador de Glucosa de Tipo 4/genética , Transportador de Glucosa de Tipo 4/metabolismo , Hipoglucemiantes/farmacología , Hipoglucemiantes/uso terapéutico , Insulina/sangre , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Ratones Endogámicos C57BL , ARN Mensajero/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Paeoniflorin(PF), extracted from the root peeled of Paeonia lactiflora Pall(Family: Ranunculaceae), has therapeutic potential in many animal models of inflammatory diseases. AIM OF THE STUDY: Although the anti-inflammatory efficacy of PF has been well illustrated in several animal models, whether it could attenuate diabetic nephropathy and detailed mechanisms are still obscure. Till now, accumulating evidence has proposed the pivotal role of toll-like receptors (TLRs) in renal inflammation in diabetic patients. In this setting, the current study aimed to investigate the effects and underlying mechanism of PF on high glucose-induced activation of toll like-receptor 2 (TLR2) signaling in macrophages. MATERIALS AND METHODS: Bone marrow-derived macrophages (BMDM) were isolated from male Tlr2tm1kir (TLR2-/-) mice and wild-type littermates (C57BL/6JWT). The level of TLR2 and activation of downstream signaling were evaluated in response to 30mmol/L high glucose (HG)-containing medium. Macrophages behaviors, which include cell viability, migration and inflammatory cytokines production, were also determined. RESULTS: PF suppressed HG-induced production of TLR2, activation of downstream signaling and synthesis of inducible nitric oxide synthase (iNOS). PF could further inhibit MyD88-dependent pathway in HG-induced models in which TLR2 was knocked out. Moreover, deletion of TLR2 inhibited the HG-induced activation of MyD88-dependent pathway, but not TIR domain containing adapter inducing interferon-ß (Trif) signal pathway in BMDMs. As HG stimulation polarizes macrophages into M1 phenotype, treatment of PF or knockout of TLR2 significantly reduces M1 markers on the membrane of macrophages. Additionally, levels of inflammatory cytokines and iNOS were remarkably reduced in response to PF or TLR2 deficiency. CONCLUSION: Collectively, these data demonstrated that HG activated macrophages primarily through TLR2-dependent mechanisms which aggravated the severity of renal inflammation and eventually contributed to DN. Additionally, PF might be applied as a potential therapeutic agent in the battle against progressive DN.
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Antiinflamatorios/farmacología , Glucosa/farmacología , Glucósidos/farmacología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Monoterpenos/farmacología , Receptor Toll-Like 2/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Relación Dosis-Respuesta a Droga , Mediadores de Inflamación/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Factor 88 de Diferenciación Mieloide/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenotipo , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Lycium barbarum L., popularly known as "Goji berry", a classic of Traditional Chinese Medicine has long been used to treat ocular diseases and cardiovascular diseases. Recently, the photoreceptor cell protection of Lycium barbarum polysaccharides (LBP), a water extract from Lycium barbarum L. has received more attention. The present study was designed to investigate the effect of LBP on N-methyl-N-nitrosourea (MNU)-induced photoreceptor cell apoptosis, and the involvement of the poly (ADP-ribose) polymerase (PARP) and caspase. MATERIALS AND METHODS: Photoreceptor cell injury was induced in male Sprague-Dawley rats by an intraperitoneal injection of MNU 60mg/kg. Seven days prior to MNU injection, LBP were intragastrical administered daily, rats were sacrificed at 24h and 7 days after MNU injection. Retinal morphologies, photoreceptor cells apoptosis, and protein expression were evaluated at 24h and 7 days after MNU injection. RESULTS: Morphologically, the outer nuclear layer was well preserved in the LBP-treated rat retinas throughout the experimental period. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling (TUNEL) assays showed that LBP could significantly suppress the loss of photoreceptor cells, as determined by the photoreceptor cell ratio at the central retina 24h and 7 days after MNU administration. Western-blot analysis demonstrated the expression levels of procaspase-9, -7, -3 and cleaved caspase-9, -7, -3 were upregulated, and PARP were downregulated both 24h and 7 days after MNU injection. LBP treatment significantly decreased protein levels of procaspase and cleaved caspase, increased the level of PARP and cleaved PARP on 24h and 7 days. CONCLUSIONS: LBP inhibits MNU-induced rat photoreceptor cell apoptosis and protects retinal structure via the regulation of the expressions of PARP and caspase.
Asunto(s)
Caspasas/metabolismo , Medicamentos Herbarios Chinos/farmacología , Lycium/química , Metilnitrosourea , Células Fotorreceptoras de Vertebrados/efectos de los fármacos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Sustancias Protectoras/farmacología , Degeneración Retiniana/prevención & control , Animales , Apoptosis/efectos de los fármacos , Citoprotección , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/aislamiento & purificación , Activación Enzimática , Masculino , Células Fotorreceptoras de Vertebrados/enzimología , Células Fotorreceptoras de Vertebrados/patología , Fitoterapia , Plantas Medicinales , Sustancias Protectoras/aislamiento & purificación , Ratas Sprague-Dawley , Degeneración Retiniana/inducido químicamente , Degeneración Retiniana/enzimología , Degeneración Retiniana/patología , Transducción de Señal/efectos de los fármacos , Factores de TiempoRESUMEN
In situ gel-forming system as local drug delivery system in dermal traumas has generated a great interest. Accumulating evidence shows that antimicrobial peptides play pivotal roles in the process of wound healing. Here in this study, to explore the potential application of antimicrobial peptide in wound healing, biodegradable poly(L-lactic acid)-Pluronic L35-poly(L-lactic acid) (PLLA-L35-PLLA) was developed at first. Then based on this polymer, an injectable in situ gel-forming system composed of human antimicrobial peptides 57 (AP-57) loaded nanoparticles and thermosensitive hydrogel was prepared and applied for cutaneous wound healing. AP-57 peptides were enclosed with biocompatible nanoparticles (AP-57-NPs) with high drug loading and encapsulation efficiency. AP-57-NPs were further encapsulated in a thermosensitive hydrogel (AP-57-NPs-H) to facilitate its application in cutaneous wound repair. As a result, AP-57-NPs-H released AP-57 in an extended period and exhibited quite low cytotoxicity and high anti-oxidant activity in vitro. Moreover, AP-57-NPs-H was free-flowing liquid at room temperature, and can form non-flowing gel without any crosslink agent upon applied on the wounds. In vivo wound healing assay using full-thickness dermal defect model of SD rats indicated that AP-57-NPs-H could significantly promote wound healing. At day 14 after operation, AP-57-NPs-H treated group showed nearly complete wound closure of 96.78 ± 3.12%, whereas NS, NPs-H and AP-57-NPs group recovered by about 68.78 ± 4.93%, 81.96 ± 3.26% and 87.80 ± 4.62%, respectively. Histopathological examination suggested that AP-57-NPs-H could promote cutaneous wound healing through enhancing granulation tissue formation, increasing collagen deposition and promoting angiogenesis in the wound tissue. Therefore, AP-57-NPs-H might have potential application in wound healing.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/administración & dosificación , Proteínas de Unión al ADN/administración & dosificación , Hidrogeles/química , Ácido Láctico/química , Nanopartículas/química , Poloxámero/química , Polímeros/química , Cicatrización de Heridas/efectos de los fármacos , Animales , Péptidos Catiónicos Antimicrobianos/farmacología , Supervivencia Celular , Proteínas de Unión al ADN/farmacología , Liberación de Fármacos , Estabilidad de Medicamentos , Células HEK293 , Humanos , Masculino , Peso Molecular , Tamaño de la Partícula , Poliésteres , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Propiedades de Superficie , TemperaturaRESUMEN
The use of crude kerosene as a dietary supplement in boarding schools has been a common practice in east Africa and other countries for many years, with the belief of it reducing the sex drive (libido) at the pubertal stage. There is however no scientific basis for this belief. The present study aimed at using a rat animal model to investigate the effects of crude kerosene on serum testosterone levels, aggression and its possible toxic effects. Fifteen male albino rats of approximately similar age and average weights were put into three groups of five animals each; the control group (placebo), low kerosene dose (10 µl/day) group and high kerosene dose (300 µl/day) group. ELISA was used to determine the serum testosterone levels. During treatment, changes in aggression were observed and noted. Liver toxicity was determined using enzyme assays, total protein and albumin while renal toxicity was monitored using serum creatinine levels. A full hemogram was conducted to determine hematological effects. Various tissue biopsies were obtained and examined using histopathological techniques for evidence of toxicity. Contrary to the common belief, our findings showed an overall increase of serum testosterone levels of up to 66% in the low dose and 75% in the high dose groups, with an increasing trend by the end of the study. The high dose group showed significantly increased levels of white blood cells (WBC) (p = 0.036), red blood cells (RBC) (p = 0.025), hematocrit (HCT) (p = 0.03), red cell distribution width (p = 0.028) and platelets (p = 0.017). The histological results of the stomach indicated chronic gastritis.
RESUMEN
BACKGROUND: Ketamine is a controlled substance and often illegally used as a recreational drug primarily by young adults. Increasing ketamine abusers associated with lower urinary tract symptoms have been reported at hospitals in recent years. Here we used a murine model to explore the changes of bladder in order to elucidate its pathogenesis. METHODS: ICR mice were randomly distributed into control and ketamine groups and received daily intraperitoneal injection of saline and ketamine (30 mg/kg), respectively. The bladders were excised and processed for histology at 4, 8 and 12 weeks. Tryptase and E-cadherin were investigated by immunohistochemistry in bladder tissues from ketamine-treated and control mice to assess the mast cell activation and junction protein expression. RESULTS: After ketamine treatment, the bladder changed to be hyperemic, inflamed, and with more fissures in mucosa. Compared with control group, the number of tryptase-positive mast cells significantly increased, which was 6.98 ± 2.89 and 23.00 ± 6.48 cells per field (100×) at 8 and 12 weeks, respectively (P = 0.016 and P = 0.003, respectively). Additionally, the expression of E-cadherin in ketamine-treated mice bladder tissue was significantly lower than that in the control tissues, P < 0.001. CONCLUSIONS: Increased mast cells in bladder wall and downregulated expression of E-cadherin junction protein in epithelial cells were probably associated with interstitial inflammation and fissures in mucosa. It implied that ketamine induced an interstitial cystitis.
RESUMEN
AIMS: ß-Adrenoceptors modulate acute wound healing; however, few studies have shown the effects of ß-adrenoceptor blockade on chronic wounds. Therefore, this study investigated the effect of ß1-/ß2-adrenoceptor blockade in wound healing of pressure ulcers. MAIN METHODS: Male mice were daily treated with propranolol (ß1-/ß2-adrenoceptor antagonist) until euthanasia. One day after the beginning of treatment, two cycles of ischemia-reperfusion by external application of two magnetic plates were performed in skin to induce pressure ulcer formation. KEY FINDINGS: Propranolol administration reduced keratinocyte migration, transforming growth factor-ß protein expression, re-epithelialization, and necrotic tissue loss. Neutrophil number and neutrophil elastase protein expression were increased in propranolol-treated group when compared with control group. Propranolol administration delayed macrophage mobilization and metalloproteinase-12 protein expression and reduced monocyte chemoattractant protein-1 protein expression. Myofibroblastic differentiation, angiogenesis, and wound closure were delayed in the propranolol-treated animals. Propranolol administration increased neo-epidermis thickness, reduced collagen deposition, and enhanced tenascin-C expression resulting in the formation of an immature and disorganized collagenous scar. SIGNIFICANCE: ß1-/ß2-Adrenoceptor blockade delays wound healing of ischemia-reperfusion skin injury through the impairment of the re-epithelialization and necrotic tissue loss which compromise wound inflammation, dermal reconstruction, and scar formation.