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1.
Int J Mol Sci ; 25(2)2024 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-38255858

RESUMO

Cirsium japonicum is a medicinal plant that has been used due to its beneficial properties. However, extensive information regarding its therapeutic potential is scarce in the scientific literature. The antioxidant and anti-inflammatory potential of polyphenols derived from the Cirsium japonicum extracts (CJE) was systematically analyzed. High-performance liquid chromatography (HPLC) with mass spectrometry (MS) was used to examine the compounds in CJE. A total of six peaks of polyphenol compounds were identified in the extract, and their MS data were also confirmed. These bioactive compounds were subjected to ultrafiltration with LC analysis to assess their potential for targeting cyclooxygenase-2 (COX2) and DPPH. The outcomes showed which primary compounds had the highest affinity for binding both COX2 and DPPH. This suggests that components that showed excellent binding ability to DPPH and COX2 can be considered significant active substances. Additionally, in vitro analysis of CJE was carried out in macrophage cells after inducing inflammation with lipopolysaccharide (LPS). As a result, it downregulated the expression of two critical pro-inflammatory cytokines, COX2 and inducible nitric oxide synthase (iNOS). In addition, we found a solid binding ability through the molecular docking analysis of the selected compounds with inflammatory mediators. In conclusion, we identified polyphenolic compounds in CJE extract and confirmed their potential antioxidant and anti-inflammatory effects. These results may provide primary data for the application of CJE in the food and pharmaceutical industries with further analysis.


Assuntos
Antioxidantes , Cirsium , Antioxidantes/farmacologia , Ciclo-Oxigenase 2 , Simulação de Acoplamento Molecular , Anti-Inflamatórios/farmacologia , Polifenóis/farmacologia , Extratos Vegetais/farmacologia
2.
Molecules ; 29(3)2024 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-38338437

RESUMO

This study aimed to isolate the proteolytic fraction from the silkworm thorn fruit (Cudrania tricuspidata) through ethanol precipitation at different ratios, and to determine its proteolytic activity and optimal activity conditions. Furthermore, the hydrolysis characteristics and antioxidant activity of soy protein isolate (SPI) and whey protein concentrate (WPC) hydrolyzates obtained through the enzymatic hydrolysis of freeze-dried silkworm thorn fruit powder (SF) were evaluated. For isolation and partial purification of proteolytic fraction, the water-solubilized fraction of the silkworm thorn fruit was purified through ethanol precipitation at four different ratios of 1:1, 1:2, 1:4, and 1:6 (v/v). The protein recovery rate, caseinolytic activity, protein pattern, and optimal activity (pH, temperature, and inhibitors) of fractional ethanol precipitate obtained from the silkworm thorn fruit (ESF) were evaluated. The proteolytic fraction obtained from silkworm thorn fruit exhibited a major protein band around 65-70 kDa and showed the highest proteolytic activity at a 1:4 ratio of ethanol precipitation (p < 0.05). The optimal activity of the measured enzyme fraction was determined to be at pH 9.0 and 50 °C, and the proteolytic activity of ESF was almost inhibited by phenyl methyl sulphonyl fluoride (PMSF, 2 mM), a serine protease inhibitor. Compared to Alcalase and papain, extensively used as commercial enzymes, the silkworm thorn fruit powder was less effective in hydrolyzing SPI and WPC. Nevertheless, SPI and WPC hydrolyzates mediated with silkworm thorn fruit powder showed even better antioxidant activities than those mediated with Alcalase and papain. Thus, our results show the potential application of silkworm thorn fruit as a novel source of plant protease for producing human-grade protein hydrolyzates.


Assuntos
Bombyx , Maclura , Animais , Humanos , Hidrólise , Bombyx/metabolismo , Papaína/metabolismo , Frutas/metabolismo , Pós , Peptídeo Hidrolases/metabolismo , Proteínas do Soro do Leite , Proteínas de Soja , Subtilisinas/metabolismo , Etanol
3.
Curr Issues Mol Biol ; 45(4): 3279-3290, 2023 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-37185738

RESUMO

The objective of this study was to elucidate the effect of cattle breed on carcass and meat chemical composition, fatty acid profiles, and lipid-metabolism-related genes. For this study, same-age Hanwoo and Chikso steers (n = 6 per breed) reared under identical conditions were used. Immediately after slaughter, muscle tissues were collected for analysis of mRNA expression. At 24 h post-mortem, the carcasses were assessed for carcass traits (marbling score, meat yield, etc.), and meat quality and fatty acid profiles in the longissimus lumborum (LL) and semimembranosus (SM) muscles. The results showed that no differences in the slaughter weight, dressing rate, back-fat thickness, trimmed fat, and total meat yield occurred between the two breeds (p > 0.05). However, Hanwoo cattle had a higher marbling score, intramuscular fat (IMF) content, and expression level of lipid-metabolism-related genes such as lipoprotein lipase, peroxisome proliferator-activated receptor gamma, and fatty acid binding protein 4, compared with Chikso (p < 0.05). Contrastingly, Chikso had a higher total unsaturated fatty acid content and expression level of stearoyl CoA desaturase 1 (p < 0.05). It may be said that the difference in the expression levels of lipid-metabolism-related genes could be the molecular factors underlying IMF deposition and fatty acid profile differences in the beef from the two breeds.

4.
Vet Ophthalmol ; 26(5): 440-445, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37565779

RESUMO

OBJECTIVE: To compare tear film (TF) osmolarity measured using TearLab® and I-PEN® osmometers in the same dogs without any ocular surface disease. ANIMAL STUDIED: Fifty-two dogs (98 eyes) of different breeds were evaluated. PROCEDURES: Tear film (TF) osmolarity was evaluated at 2-min intervals. The test was randomly determined, and single measurements were performed using each osmometer. Subsequently, complete ophthalmologic examinations were performed based on Schirmer tear test-1 (STT-1) analysis, tear film breakup time (TFBUT), and slit-lamp biomicroscopy. For each osmometer, the mean ± standard deviation of the TF osmolarity was calculated, and a paired Student's t-test was used to compare the values obtained. Pearson correlation analysis was performed to assess the association between osmolarity and other values such as STT-1, TFBUT, and age. RESULTS: Tear film osmolarity determined using TearLab® (340.42 ± 15.87 mOsm/L) and I-PEN® (321.58 ± 17.39 mOsm/L) were significantly different (p < .001). However, statistical significance could not be confirmed between osmolarity and other values, such as STT-1, TFBUT, and age. CONCLUSIONS: In dogs, the TF osmolarity values obtained using TearLab® tend to be higher than those obtained using I-PEN®, contrary to that observed in humans. These findings can serve as a reference for establishing normal values for each osmometer for clinical use in measuring TF osmolarity in dogs.


Assuntos
Doenças do Cão , Lacerações , Animais , Cães , Olho , Lacerações/veterinária , Concentração Osmolar , Osmometria/veterinária , Microscopia com Lâmpada de Fenda , Lágrimas
5.
Int J Mol Sci ; 24(11)2023 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-37298192

RESUMO

Cancer is a widespread but dangerous disease that can strike anyone and is the second 1leading cause of death worldwide. Prostate cancer, in particular, is a prevalent cancer that occurs in men, and much research is being done on its treatment. Although chemical drugs are effective, they have various side effects, and accordingly, anticancer drugs using natural products are emerging. To date, many natural candidates have been discovered, and new drugs are being developed as drugs to treat prostate cancer. Representative candidate compounds that have been studied to be effective in prostate cancer include apigenin, acacetin and tangeretin of the flavone family among flavonoids. In this review, we look at the effects of these three flavones on prostate cancer cells via apoptosis in vitro and in vivo. Furthermore, in addition to the existing drugs, we suggest the three flavones and their effectiveness as natural anticancer agents, a treatment model for prostate cancer.


Assuntos
Antineoplásicos , Flavonas , Neoplasias da Próstata , Masculino , Humanos , Flavonas/farmacologia , Flavonas/química , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Apoptose , Apigenina/farmacologia , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico
6.
Molecules ; 28(14)2023 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-37513381

RESUMO

Pet owners think of their animals as part of their family, which further promotes the growth of the pet food market, encouraging pet owners to select nutritious, palatable, and high-quality foods for pets. Therefore, the evaluation of taste and volatile compounds in pet foods is essential to improve palatability. In this study, the sensory characteristics of taste and odor compounds in 10 commercially available dry dog foods were investigated using electronic tongue (E-tongue), electronic nose (E-nose), gas chromatography-mass spectrometry (GC-MS), and gas chromatography-olfactometry (GC-O). Dry dog foods were separated based on the sensory properties of taste and volatile compounds through the multivariate analysis of integrated results of the E-tongue and E-nose. A total of 67 odor active compounds were detected through GC-MS and GC-O, and octanal, nonanal, 2-pentyl furan, heptanal, and benzaldehyde were identified as key odor compounds which may have positive effects on food intake. The multivariate analysis was used to classify samples based on key odor compounds. Volatile compounds responsible for aroma properties of samples were evaluated using GC-O and multivariate analysis in this present study for the first time. These results are expected to provide fundamental data for sensory evaluation in producing new dog foods with improved palatability.


Assuntos
Olfato , Compostos Orgânicos Voláteis , Cães , Animais , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ração Animal/análise , Paladar , Compostos Orgânicos Voláteis/análise , Odorantes/análise , Olfatometria/métodos , Eletrônica , Nariz Eletrônico
7.
Exp Dermatol ; 31(6): 931-935, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35181944

RESUMO

Recently, light-emitting diode (LED)-based devices have emerged as effective and safe tools for the treatment of photoaged skin. However, few studies have been conducted to elucidate the underlying mechanism behind the effect on photoageing of LED light. In this study, we induced photoageing of human dermal fibroblasts (HDFs) with Ultraviolet B (UVB) irradiation and evaluated the ability of 590-nm LED radiation to induce recovery from oxidative stress, restore collagen formation and regulate inflammatory changes. Photoageing was induced in cultured human dermal fibroblasts (HDFs) using UVB irradiation of 50 mJ/cm2 . Then, the photoaged HDFs were irradiated with LED using a custom-built 590-nm LED device which emits light with an intensity of 38 mW/cm2 (irradiated for 900 s with 34.2 J/cm2 of total energy). LED irradiation significantly attenuated UVB-induced reactive oxygen species generation and UVB-induced phosphorylation of JNK, c-Fos and c-Jun. In addition, the procollagen levels were recovered significantly, and MMP-9 levels were significantly suppressed after LED irradiation. The UVB-induced phosphorylation levels of NF-κB and pro-inflammatory enzyme COX-2 also significantly decreased. Our results suggest that 590-nm yellow light irradiation may be an effective and safe anti-oxidative and anti-inflammatory treatment modality for photoaged skin.


Assuntos
Envelhecimento da Pele , Raios Ultravioleta , Fibroblastos , Humanos , Estresse Oxidativo/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Pele/metabolismo
8.
Int J Mol Sci ; 23(16)2022 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-36012164

RESUMO

Kynurenic acid was included in the three compounds (caffeic acid, chlorogenic acid, and kynurenic acid) that showed high antioxidant and anti-inflammatory potential among the phenolic compounds contained in Gynura procumbens. In this study, the mechanism of cancer cell death induced by kynurenic acid (KYNA), which has the highest molecular binding affinity, in the gastric cancer cell line AGS was confirmed in molecular docking analysis. KYNA showed the most cancer cell death effect on AGS cells among several gastric cancer cell lines (MKN, AGS, and SNU). AGS cells were used for later experiments, and KYNA concentrations of 0, 150, 200, and 250 µM were used. KYNA inhibited cell migration and proliferation in AGS cells in a concentration-dependent manner. G2/M phase cell cycle arrest and reduction of related proteins (Cdc25C, CDK1 and CyclinB1) were confirmed in KYNA-treated AGS cells. Apoptosis of KYNA-treated AGS cells was confirmed by Annexin V/propidium iodide (PI) staining flow cytometry analysis. As a result of morphological chromatin condensation through DAPI (4',6-diamidino-2-phenylindole), intense blue fluorescence was confirmed. The mechanism of apoptosis induction of KYNA-treated AGS cells was confirmed by western blotting. In the extrinsic pathway, apoptosis induction markers FasL, Fas, and Caspase-3 and -8 were increased in a concentration-dependent manner upon KYNA treatment. In the intrinsic pathway, the expression of anti-apoptotic factors PI3K, AKT, and Bcl-xL was down-regulated, and the expression of apoptosis-inducing factors BAD, Bak, Bax, Cytochrom C, and Caspase-9 was up-regulated. Therefore, in the present study, we strongly imply that KYNA induces apoptosis in AGS gastric cancer cells. This suggests that KYNA, a natural compound, could be the basis for drug for the treatment of gastric cancer.


Assuntos
Neoplasias Gástricas , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Ácido Cinurênico/farmacologia , Ácido Cinurênico/uso terapêutico , Simulação de Acoplamento Molecular , Neoplasias Gástricas/metabolismo
9.
Int J Mol Sci ; 23(18)2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36142874

RESUMO

Cancer is a horrific disease that, to date, has no cure. It is caused by various factors and takes many lives. Apoptosis is a programmed cell death mechanism and if it does not function correctly in cancer cells, it can lead to severe disease. There are various signaling pathways for regulating apoptosis in cancer cells. Flavonoids are non-artificial natural bioactive compounds that are gaining attention as being capable of for inducing apoptosis in cancer cells. Among these, in this study, we focus on flavones. Flavones are a subclass of the numerous available flavonoids and possess several bioactive functions. Some of the most reported and well-known critical flavones, namely apigenin, acacetin, baicalein, luteolin, tangeretin, and wogonin, are discussed in depth in this review. Our main aim is to investigate the effects of the selected flavones on apoptosis and cell signaling pathways that contribute to death due to various types of cancers.


Assuntos
Flavonas , Neoplasias , Apigenina/farmacologia , Apoptose , Flavonas/farmacologia , Flavonoides/farmacologia , Humanos , Luteolina/farmacologia , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Transdução de Sinais
10.
Asian-Australas J Anim Sci ; 33(4): 662-669, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31480199

RESUMO

OBJECTIVE: This study was conducted to evaluate the effect of red glasswort (RG) (Salicornia herbacea L.) curing on the physicochemical, textural and sensory properties of cooked pork loin ham (M. longissimus thoracis et lumborum). METHODS: All treatments were cured with different salt and RG powder levels. RG0 treatment was prepared with only 4% NaCl (w/w) as a control, and RG25, 3% NaCl:1% RG (w/w); RG50, 2% NaCl:2% RG (w/w); RG75, 1% NaCl:3% RG (w/w); RG100, 0% NaCl:4% RG (w/w) treatments were prepared sequentially. All samples were individually vacuum packaged in polyethylene bags and stored for 7 d at 3°C±1°C. RESULTS: The results showed that as the rate of RG substitution increased, pH value, redness, myofibrillar protein solubility, and myofibrillar fragmentation index increased (p<0.05), but salt concentration and shear force decreased (p<0.05). However, there were no significant differences in cooking loss and moisture content. In terms of sensory evaluation, RG100 exhibited higher scores in tenderness and juiciness than RG0 (p<0.05). CONCLUSION: The partial substitution of NaCl by RG could improve the physicochemical properties, textural and sensory characteristics of cooked pork loin. Therefore, it is suggested that RG as a natural salt replacer could be an effective ingredient for developing low-sodium cured hams.

11.
Asian-Australas J Anim Sci ; 33(3): 501-505, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31480178

RESUMO

OBJECTIVE: The purpose of this study was to investigate the effects of aging methods (AM) i.e. dry-aging (DA) and wet-aging (WA) on the physicochemical properties and in vitro digestibility of proteins in beef short loin. METHODS: Short loins (M. longissmus lumborum), were trimmed and boned-out on the fifth day postmortem, from a total of 18 Hanwoo, which were purchased from a commercial slaughterhouse. Short loins were separated randomly grouped into one of the three treatments: control, WA (1°C, 7 days), and DA (1°C, 0.5 m/s, 85% relative humidity [RH], 30 days). RESULTS: Dry-aged beef (DAB) exhibited higher pH, water holding capacity (WHC), myofibrillar fragmentation index (MFI), and digestibility, however lower lightness, redness, and yellowness values, cooking loss, and shear force (SF), than those of wet-aged beef (WAB) (p<0.05). The myosin light chain band intensity of DAB was higher than that of control and WAB in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The in vitro digestibility of aged beef was highly (p<0.001) correlated to physicochemical properties except WHC. The correlation coefficient between AMs and WHC was higher than that between AM and SF (p<0.05) or MFI (p<0.001). A high correlation was observed between SF and MFI (p<0.001). CONCLUSION: Thus, we believe that DAB is more advantageous than WAB owing to its high digestibility and WHC and low SF.

12.
J Food Sci Technol ; 57(8): 3142-3150, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32624615

RESUMO

The objective of this study was to compare the meat quality attributes, color stability, and lipid oxidation of loin chops from finishing gilts and cull sows in a three-way crossbreeding system: landrace × large white × duroc finishing gilts (n = 20) and landrace × large white sows (n = 20). No significant differences in pH, proximate composition, or total collagen content were found between the two pig groups. However, sow loin chops exhibited different quality characteristics for color, cooking loss, protein solubility, and shear force, in which an increased cooking loss and shear force might be associated with a lower heat-soluble collagen content compared to the gilt loin chops (p < 0.05). Moreover, more rapid changes in redness and chroma during 7 days of aerobic display were observed in the sow loin chops (p < 0.05). Therefore, the results presented here highlight the possibility of issues regarding color and tenderness in sow meat compared to retail pork produced from finishing gilts.

13.
Neuroendocrinology ; 108(3): 172-189, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30537700

RESUMO

BACKGROUND/AIMS: The medial amygdala (MeA) responds to olfactory stimuli and alters reproductive physiology. However, the neuronal circuit that relays signals from the MeA to the reproductive axis remains poorly defined. This study aimed to test whether MeA kisspeptin (MeAKiss) neurons in male mice are sensitive to sexually relevant olfactory stimuli and transmit signals to alter reproductive physiology. We also investigated whether MeAKiss neurons have the capacity to elaborate glutamate and GABA neurotransmitters and potentially contribute to reproductive axis regulation. METHODS: Using female urine as a pheromone stimulus, MeAKiss neuronal activity was analysed and serum luteinizing hormone (LH) was measured in male mice. Next, using a chemogenetic approach, MeAKiss neurons were bi-directionally modulated to measure the effect on serum LH and evaluate the activation of the preoptic area. Lastly, using in situ hybridization, we identified the proportion of MeAKiss neurons that express markers for GABAergic (Vgat) and glutamatergic (Vglut2) neurotransmission. RESULTS: Male mice exposed to female urine showed a two-fold increase in the number of c-Fos-positive MeAKiss neurons concomitant with raised LH. Chemogenetic activation of MeAKiss neurons significantly increased LH in the absence of urine exposure, whereas inhibition of MeAKiss neurons did not alter LH. In situ hybridization revealed that MeAKiss neurons are a mixed neuronal population in which 71% express Vgat mRNA, 29% express Vglut2 mRNA, and 6% express both. CONCLUSIONS: Our results uncover, for the first time, that MeAKiss neurons process sexually relevant olfactory signals to influence reproductive hormone levels in male mice, likely through a complex interplay of neuropeptide and neurotransmitter signalling.


Assuntos
Tonsila do Cerebelo/fisiologia , Kisspeptinas/fisiologia , Hormônio Luteinizante/sangue , Neurônios/fisiologia , Feromônios/farmacologia , Administração por Inalação , Tonsila do Cerebelo/efeitos dos fármacos , Tonsila do Cerebelo/metabolismo , Animais , Clozapina/análogos & derivados , Clozapina/farmacologia , Ácido Glutâmico/metabolismo , Kisspeptinas/genética , Masculino , Camundongos , Camundongos Transgênicos , Feromônios/administração & dosagem , Feromônios/urina , Área Pré-Óptica/fisiologia , Proteína Vesicular 2 de Transporte de Glutamato/metabolismo , Proteínas Vesiculares de Transporte de Aminoácidos Inibidores/metabolismo , Ácido gama-Aminobutírico/metabolismo
14.
J Food Sci Technol ; 56(12): 5282-5288, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31749475

RESUMO

The objectives of this study were to determine the effects of tumbling time on the processing characteristics of soy sauce marinated pork loins, and to compare its efficacy at two different tumbling temperatures. In experiment I, pork loins (M. longissimus thoracis et lumborum) were pumped to 115% (w/w) of original sample weight with soy sauce solution (75%, w/w) and tumbled for 0, 1, 3, and 5 h. In experiment II, the soy sauce pumped pork loins were tumbled at two different temperatures (- 1 and 4 °C) for 4 h. The pH, color characteristics, water-holding capacity, protein solubility, and shear force of pork loins tumbled with soy sauce marinade were determined. An increase in tumbling time increased the cooked pH and yellowness of soy sauce marinated pork loins (p < 0.05). Protein solubility, particularly that of total and salt-soluble proteins, increased with increasing tumbling time (p < 0.05), and a tumbling time of at least 3 h may guarantee excellent water-holding capacity and shear force. In addition, low tumbling temperature of - 1 °C could positively contribute to the improvement of water-holding capacity and shear force of soy sauce marinated pork loins. In conclusion, this current study indicates that the efficacy of tumbling in soy sauce marination of pork loins is affected by tumbling time and temperature, moreover, tumbling at - 1 °C for at least 3 h could be an effective method in the experimental operating condition.

15.
Asian-Australas J Anim Sci ; 31(8): 1358-1365, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29381898

RESUMO

OBJECTIVE: The effect of pre-converted nitrites from natural sources (spinach, lettuce, celery, and red beet) on color development in raw and cooked pork sausage was investigated in this study. METHODS: The pork sausage was manufactured with six treatments: NC (negative control, nitrite free), PC (positive control, 150 ppm sodium nitrite), FS (3.0% fermented spinach extracts), FL (3.0% fermented lettuce extracts), FC (3.0% fermented celery extracts), and FR (3.0% fermented red beet extracts). RESULTS: The pH value of the pre-converted nitrites groups was lower than those treated with 150 ppm sodium nitrite (p<0.05). The color values of raw and cooked pork sausage added with pre-converted nitrite showed slightly lower and/or similar lightness, lower redness, and higher yellowness values than PC. Color development (redness values) of cooked samples added with FS was higher than those of the NC and other treatments (FL, FC, and FR). Additionally, treatments with FS and FL were most effective for reducing thiobarbituric acid reactive substances and volatile basic nitrogen than the NC. CONCLUSION: Effects of natural nitrites from fermented vegetables on shelf stability of raw and cooked pork sausages were investigated. Fermented spinach extract was much more useful for maintaining the color development, but also inhibiting lipid and protein oxidation of cooked pork sausage. Therefore, pre-converted nitrite from spinach as a natural nitrite could be used as another natural nitrite source for making processed meat products.

16.
Korean J Physiol Pharmacol ; 22(3): 311-319, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29719453

RESUMO

Mitochondrial calcium overload is a crucial event in determining the fate of neuronal cell survival and death, implicated in pathogenesis of neurodegenerative diseases. One of the driving forces of calcium influx into mitochondria is mitochondria membrane potential (ΔΨm). Therefore, pharmacological manipulation of ΔΨm can be a promising strategy to prevent neuronal cell death against brain insults. Based on these issues, we investigated here whether nobiletin, a Citrus polymethoxylated flavone, prevents neurotoxic neuronal calcium overload and cell death via regulating basal ΔΨm against neuronal insult in primary cortical neurons and pure brain mitochondria isolated from rat cortices. Results demonstrated that nobiletin treatment significantly increased cell viability against glutamate toxicity (100 µM, 20 min) in primary cortical neurons. Real-time imaging-based fluorometry data reveal that nobiletin evokes partial mitochondrial depolarization in these neurons. Nobiletin markedly attenuated mitochondrial calcium overload and reactive oxygen species (ROS) generation in glutamate (100 µM)-stimulated cortical neurons and isolated pure mitochondria exposed to high concentration of Ca2+ (5 µM). Nobiletin-induced partial mitochondrial depolarization in intact neurons was confirmed in isolated brain mitochondria using a fluorescence microplate reader. Nobiletin effects on basal ΔΨm were completely abolished in K+-free medium on pure isolated mitochondria. Taken together, results demonstrate that K+ influx into mitochondria is critically involved in partial mitochondrial depolarization-related neuroprotective effect of nobiletin. Nobiletin-induced mitochondrial K+ influx is probably mediated, at least in part, by activation of mitochondrial K+ channels. However, further detailed studies should be conducted to determine exact molecular targets of nobiletin in mitochondria.

17.
Lab Invest ; 97(4): 419-431, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28165467

RESUMO

Recent studies have suggested that renal Nox is important in the progression of diabetic nephropathy. Therefore, we investigated the effect of a novel pan-NOX-inhibitor, APX-115, on diabetic nephropathy in type 2 diabetic mice. Eight- week-old db/m and db/db mice were treated with APX-115 for 12 weeks. APX-115 was administered by oral gavage at a dose of 60 mg/kg per day. To compare the effects of APX-115 with a dual Nox1/Nox4 inhibitor, db/db mice were treated with GKT137831 according to the same protocol. APX-115 significantly improved insulin resistance in diabetic mice, similar to GKT137831. Oxidative stress as measured by plasma 8-isoprostane level was decreased in the APX-115 group compared with diabetic controls. All lipid profiles, both in plasma and tissues improved with Nox inhibition. APX-115 treatment decreased Nox1, Nox2, and Nox4 protein expression in the kidney. APX-115 decreased urinary albumin excretion and preserved creatinine level. In diabetic kidneys, APX-115 significantly improved mesangial expansion, but GKT137831 did not. In addition, F4/80 infiltration in the adipose tissue and kidney decreased with APX-115 treatment. We also found that TGF-ß stimulated ROS generation in primary mouse mesangial cells (pMMCs) from wild-type, Nox1 KO, and Duox1 KO mice, but did not induce Nox activity in pMMCs from Nox2 knockout (KO), Nox4 KO, or Duox2 KO mice. These results indicate that activating Nox2, Nox4, or Duox2 in pMMCs is essential for TGF-ß-mediated ROS generation. Our findings suggest that APX-115 may be as effective or may provide better protection than the dual Nox1/Nox4 inhibitor, and pan-Nox inhibition with APX-115 might be a promising therapy for diabetic nephropathy.


Assuntos
Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/prevenção & controle , Inibidores Enzimáticos/farmacologia , NADPH Oxidases/antagonistas & inibidores , Pirazóis/farmacologia , Piridinas/farmacologia , Animais , Western Blotting , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/genética , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Isoenzimas/antagonistas & inibidores , Isoenzimas/genética , Isoenzimas/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Masculino , Células Mesangiais/efeitos dos fármacos , Células Mesangiais/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Pirazolonas , Piridonas , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta/farmacologia
18.
Exp Cell Res ; 340(1): 12-21, 2016 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-26704260

RESUMO

Many recent studies have revealed that axin is involved in numerous cellular functions beyond the negative regulation of ß-catenin-dependent Wnt signaling. Previously, an association of ectopic axin with mitochondria was observed. In an effort to investigate the relationship between axin and mitochondria, we found that axin expression suppressed cellular ATP production, which was more apparent as axin expression levels increased. Also, mitochondrial expression of axin was observed using two axin-expressing HeLa cell models: doxycycline-inducible ectopic axin expression (HeLa-axin) and axin expression enhanced by long-term treatment with XAV939 (HeLa-XAV). In biochemical analysis, axin is associated with oxidative phosphorylation (OXPHOS) complex IV and is involved in defects in the assembly of complex IV-containing supercomplexes. Functionally, axin expression reduced the activity of OXPHOS complex IV and the oxygen consumption rate (OCR), suggesting axin-mediated mitochondrial dysfunction. Subsequent studies using various inhibitors of Wnt signaling showed that the reduction in cellular ATP levels was weaker in cases of ICAT protein expression and treatment with iCRT3 or NSC668036 compared with XAV939 treatment, suggesting that XAV939 treatment affects ATP synthesis in addition to suppressing Wnt signaling activity. Axin-mediated regulation of mitochondrial function may be an additional mechanism to Wnt signaling for regulation of cell growth.


Assuntos
Trifosfato de Adenosina/biossíntese , Proteína Axina/genética , Proteína Axina/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Trifosfato de Adenosina/metabolismo , Proliferação de Células/efeitos dos fármacos , Doxiciclina/farmacologia , Células HeLa , Humanos , Mitocôndrias/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real
19.
J Sci Food Agric ; 97(15): 5343-5351, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28493474

RESUMO

BACKGROUND: The present study aimed to evaluate the effects of heat stress and probiotic supplementation on protein functionality and oxidative stability of ground chicken leg during display storage. Two hundred and forty, 1-day-old male chicks (5 birds per pen) were subjected to four treatments in a 2 (thermoneutral condition at 21 °C and cyclic heat stress at 32-21-32 °C for 10 h day-1 ) × 2 (regular diet with 0 or 0.25 g kg-1 Bacillus subtilis) factorial design. Chickens were harvested at day 46, and pairs of whole legs were collected at 1 day postmortem. The chicken legs were deboned, ground, tray-packaged with oxygen-permeable film, and displayed for 3 days. RESULTS: Heat stress and probiotic supplementation had no impact on pH, water-holding capacity, color, protein functionality, lipid lipolysis and lipid/protein oxidation stability (P > 0.05). Display storage increased the pH and lipid oxidation of ground chicken legs (P < 0.05). In addition, protein oxidation occurred during display storage, as determined via an increased carbonyl group (P = 0.0109) and reduced thiol group (P < 0.0001). CONCLUSION: The results of the present study indicate that chronic heat stress and probiotic supplementation had no practical adverse impact on protein functionality and oxidative stability of ground chicken leg meat. © 2017 Society of Chemical Industry.


Assuntos
Ração Animal/análise , Galinhas/microbiologia , Carne/análise , Probióticos/administração & dosagem , Proteínas/química , Animais , Galinhas/fisiologia , Suplementos Nutricionais/análise , Armazenamento de Alimentos , Temperatura Alta , Lipídeos/química , Masculino , Músculo Esquelético , Oxirredução , Proteínas/metabolismo
20.
Asian-Australas J Anim Sci ; 30(2): 254-261, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27488843

RESUMO

OBJECTIVE: The effects of aging and freezing/thawing sequence on color, physicochemical, and enzymatic characteristics of two beef muscles (Mm. gluteus medius, GM and biceps femoris, BF) were evaluated. METHODS: Beef muscles at 3 d postmortem were assigned to four different combinations of aging and freezing/thawing sequence as follows; aging at 2°C for 3 wk (A3, never-frozen control), freezing at -28°C for 2 wk then thawing (F2, frozen/thawed-only), aging at 2°C for 3 wk, freezing at -28°C for 2 wk then thawing (A3F2), and freezing at -28°C for 2 wk, thawing then further aging at 2°C for 3 wk (F2A3). RESULTS: No significant interactions between different aging/freezing/thawing treatments and muscle type on all measurements were found. Postmortem aging, regardless of aging/freezing/thawing sequence, had no impact on color stability of frozen/thawed beef muscles (p<0.05). F2A3 resulted in higher purge loss than F2 and A3F2 treatments (p<0.05). A3F2 and F2A3 treatments resulted in lower shear force of beef muscles compared to F2 (p<0.05). Although there was no significant difference in glutathione peroxidase (GSH-Px) activity, F2A3 had the highest ß-N-acetyl glucominidase (BNAG) activity in purge, but the lowest BNAG activity in muscle (p<0.05). GM muscle exhibited higher total color changes and purge loss, and lower GSH-Px activity than BF muscle. CONCLUSION: The results from this present study indicate that different combinations of aging/freezing/thawing sequence would result in considerable impacts on meat quality attributes, particularly thaw/purge loss and tenderness. Developing a novel freezing strategy combined with postmortem aging will be beneficial for the food/meat industry to maximize its positive impacts on tenderness, while minimizing thaw/purge loss of frozen/thawed meat.

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