RESUMEN
The selective interaction of cytochrome c (Cyt c) with cardiolipin (CL) is involved in mitochondrial membrane permeabilization, an essential step for the release of apoptosis activators. The structural basis and modulatory mechanism are, however, poorly understood. Here, we report that Cyt c can induce CL peroxidation independent of reactive oxygen species, which is controlled by its redox states. The structural basis of the Cyt c-CL binding was unveiled by comprehensive spectroscopic investigation and mass spectrometry. The Cyt c-induced permeabilization and its effect on membrane collapse, pore formation, and budding are observed by confocal microscopy. Moreover, cytochrome c oxidase dysfunction is found to be associated with the initiation of Cyt c redox-controlled membrane permeabilization. These results verify the significance of a redox-dependent modulation mechanism at the early stage of apoptosis, which can be exploited for the design of cytochrome c oxidase-targeted apoptotic inducers in cancer therapy.
Asunto(s)
Citocromos c , Espectrometría Raman , Citocromos c/química , Citocromos c/metabolismo , Citocromos c/farmacología , Complejo IV de Transporte de Electrones/metabolismo , Oxidación-Reducción , Cardiolipinas/química , Cardiolipinas/metabolismo , Cardiolipinas/farmacología , Membranas Mitocondriales/metabolismo , ApoptosisRESUMEN
Background and Objectives: Cancer is the second-most-important deadly disease in the world, leading to severe socioeconomic consequences and posing a public threat. Consequently, breast and colorectal cancers are significant cancer types that affect women and men more commonly, respectively. Treatment failure or recurrent diseases frequently occur due to resistance, in addition to the side effects of the currently available anticancer agents. Therefore, in this study, herbal melanin anticancer activity was investigated against human breast adenocarcinoma (MDA-MB-231) and human colorectal (HCT 116) cell proliferation and the expression of downregulated anti-apoptotic proteins and upregulated pro-apoptotic p53. Materials and Methods: MDA-MB-231 and HCT 116 cells were monitored for their real-time proliferation properties using Xcelligence. Herbal melanin of various concentrations significantly inhibited MDA-MB-231 and HCT 116 cell proliferation. Then, the expression of proapoptotic and anti-apoptotic proteins such as p53, Bcl-2 and Bcl-xl was studied using Western blotting. Results: The Bcl-2 and Bcl-xl expressions were downregulated, while the p53 expression was upregulated after treatment with herbal melanin. Similarly, the expression of apoptotic proteins such as Bcl-2, Bcl-xl, XIAP, Survivin, Bid, Bax, p53, Cytochrome C, PARP genes and mRNA was studied after herbal melanin treatment using real-time PCR, which revealed the downregulation of Bcl-2, Bcl-xl, XIAP and Survivin and the upregulation of Bid, Bax, p53, Cytochrome C and PARP apoptotic protein expression. Also, caspase 3 and 9 expressions were monitored after the treatment with herbal melanin, which revealed the upregulation of both the MDA-MB-231 and HCT 116 cell types. Conclusions: Overall, herbal melanin can be used as an alternative anticancer agent against the MDA-MB-231 and HCT 116 cell types.
Asunto(s)
Antineoplásicos , Neoplasias de la Mama , Femenino , Humanos , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Reguladoras de la Apoptosis/farmacología , Proteínas Reguladoras de la Apoptosis/uso terapéutico , Células HCT116 , Proteína p53 Supresora de Tumor/genética , Survivin/metabolismo , Survivin/farmacología , Survivin/uso terapéutico , Melaninas/metabolismo , Melaninas/farmacología , Melaninas/uso terapéutico , Apoptosis , Proteína X Asociada a bcl-2/genética , Citocromos c/metabolismo , Citocromos c/farmacología , Citocromos c/uso terapéutico , Inhibidores de Poli(ADP-Ribosa) Polimerasas/farmacología , Inhibidores de Poli(ADP-Ribosa) Polimerasas/uso terapéutico , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proliferación Celular , Antineoplásicos/uso terapéutico , Neoplasias de la Mama/genética , Línea Celular TumoralRESUMEN
BACKGROUND: Steroid-induced avascular necrosis of the femoral head (SANFH) is characterized by osteoblast apoptosis, leading to a loss of bone structure and impaired hip joint function. It has been demonstrated that erythropoietin (EPO) performs a number of biological roles. OBJECTIVE: We examined the effects of EPO on SANFH and its regulation of the STAT1-caspase 3 signaling pathway. METHOD: In vitro, osteoblasts were treated with dexamethasone (Dex) or EPO. We identified the cytotoxicity of EPO by CCK-8, the protein expression of P-STAT1, cleaved-caspase9, cleaved-caspase3, Bcl-2, BAX, and cytochrome c by Western blotting, and evaluated the apoptosis of osteoblasts by flow cytometry. In vivo, we analyzed the protective effect of EPO against SANFH by hematoxylin and eosin (H&E), Immunohistochemical staining, and Micro-computed tomography (CT). RESULTS: In vitro, EPO had no apparent toxic effect on osteoblasts. In Dex-stimulated cells, EPO therapy lowered the protein expression of BAX, cytochrome c, p-STAT1, cleaved-caspase9, and cleaved-caspase3 while increasing the expression of Bcl-2. EPO can alleviate the apoptosis induced by Dex. In vivo, EPO can lower the percentage of empty bone lacunae in SANFH rats. CONCLUSION: The present study shows that EPO conferred beneficial effects in rats with SANFH by inhibiting STAT1-caspase 3 signaling, suggesting that EPO may be developed as a treatment for SANFH.
Asunto(s)
Eritropoyetina , Necrosis de la Cabeza Femoral , Ratas , Animales , Caspasa 3/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Necrosis de la Cabeza Femoral/inducido químicamente , Necrosis de la Cabeza Femoral/tratamiento farmacológico , Necrosis de la Cabeza Femoral/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Microtomografía por Rayos X , Apoptosis , Transducción de Señal , Osteoblastos/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Eritropoyetina/farmacología , Esteroides/efectos adversosRESUMEN
BACKGROUND: Medical therapies can cause cardiotoxicity. Chloroquine (QC) and hydroxychloroquine (HQC) are drugs used in the treatment of malaria and skin and rheumatic disorders. These drugs were considered to help treatment of coronavirus disease (COVID-19) in 2019. Despite the low cost and availability of QC and HQC, reports indicate that this class of drugs can cause cardiotoxicity. The mechanism of this event is not well known, but evidence shows that QC and HQC can cause cardiotoxicity by affecting mitochondria and lysosomes. METHODS: Therefore, our study was designed to investigate the effects of QC and HQC on heart mitochondria. In order to achieve this aim, mitochondrial function, reactive oxygen species (ROS) level, mitochondrial membrane disruption, and cytochrome c release in heart mitochondria were evaluated. Statistical significance was determined using the one-way and two-way analysis of variance (ANOVA) followed by post hoc Tukey to evaluate mitochondrial succinate dehydrogenase (SDH) activity and cytochrome c release, and Bonferroni test to evaluate the ROS level, mitochondrial membrane potential (MMP) collapse, and mitochondrial swelling. RESULTS: Based on ANOVA analysis (one-way), the results of mitochondrial SDH activity showed that the IC50 concentration for CQ is 20 µM and for HCQ is 50 µM. Based on two-way ANOVA analysis, the highest effect of CQ and HCQ on the generation of ROS, collapse in the MMP, and mitochondrial swelling were observed at 40 µM and 100 µM concentrations, respectively (p < 0.05). Also, the highest effect of these two drugs has been observed in 60 min (p < 0.05). The statistical results showed that compared to CQ, HCQ is able to cause the release of cytochrome c from mitochondria in all applied concentrations (p < 0.05). CONCLUSIONS: The results suggest that QC and HQC can cause cardiotoxicity which can lead to heart disorders through oxidative stress and disfunction of heart mitochondria.
Asunto(s)
COVID-19 , Hidroxicloroquina , Humanos , Hidroxicloroquina/toxicidad , Cloroquina/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Cardiotoxicidad/etiología , Cardiotoxicidad/tratamiento farmacológico , Citocromos c/metabolismo , Citocromos c/farmacología , Tratamiento Farmacológico de COVID-19 , MitocondriasRESUMEN
Toosendanin (TSN) is an active compound from the fruit of Melia toosendan Sieb et Zucc. TSN has been shown to have broad-spectrum anti-tumour activities in human cancers. However, there are still many gaps in the knowledge of TSN on canine mammary tumours (CMT). CMT-U27 cells were used to select the optimal acting time and best concentration of TSN to initiate apoptosis. Cell proliferation, cell colony formation, cell migration and cell invasion were analysed. The expression of apoptosis-related genes and proteins were also detected to explore the mechanism of action of TSN. A murine tumour model was established to detect the effect of TSN treatments. The results showed that TSN decreased cell viability of migration and invasion, altered CMT-U27 cell morphology, and inhibited DNA synthesis. TSN-induced cell apoptosis by upregulating BAX, cleaved caspase-3, cleaved caspase-9, p53 and cytochrome C (cytosolic) protein expression, and downregulating Bcl-2 and cytochrome C (mitochondrial) expression. In addition, TSN increased the mRNA transcription levels of cytochrome C, p53 and BAX, and decreased the mRNA expression of Bcl-2. Furthermore, TSN inhibited the growth of CMT xenografts by regulating the expression of genes and proteins activated by the mitochondrial apoptotic pathway. In conclusion, TSN effectively inhibited cell proliferation, migration and invasion activity, as well as induced CMT-U27 cell apoptosis. The study provides a molecular basis for the development of clinical drugs and other therapeutic options.
Asunto(s)
Enfermedades de los Perros , Medicamentos Herbarios Chinos , Neoplasias , Humanos , Animales , Perros , Ratones , Proteína X Asociada a bcl-2/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Proteína p53 Supresora de Tumor , Enfermedades de los Perros/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Apoptosis , Neoplasias/tratamiento farmacológico , Neoplasias/veterinaria , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero , Línea Celular TumoralRESUMEN
Spinal cord injury (SCI) is a severe traumatic event, but without any established effective treatment because of the irreversible neuronal death. Here, we investigated the role of miR-222-3p in neuronal apoptosis following SCI. Rat SCI models and neuron hypoxia models were accordingly established. The Bbc3, Bim, Bcl-2, Bax, cleaved-caspase 3, cleaved-caspase 9, Cytochrome c, and miR-222-3p expression levels were examined by Western blotting and real-time reverse transcription polymerase chain reaction (RT-qPCR). The possible association between miR-222-3p and Bbc3/Bim was analyzed by dual-luciferase assay. The neuron viability was assessed by Cell Counting Kit-8 assay and Nissl's staining. Live cell staining was performed to detect the mitochondrial membrane potential and neuronal apoptosis. Rat locomotor function was assessed using the Basso-Beattie-Bresnahan scores. Cytochrome c was outflowed from the mitochondria after SCI or hypoxia treatment, and Bbc3, Bim, Bax, cleaved-caspase 9, and cleaved-caspase 3 were significantly upregulated, while Bcl-2 and miR-222-3p were decreased remarkably. Meanwhile, neuronal cell viability was significantly inhibited. Treatment of miR-222-3p significantly suppressed the Cytochrome c efflux and neuronal apoptosis and improved neuronal cell viability and motor function in SCI rats. Moreover, we found that Bbc3 and Bim were the direct targets of miR-222-3p. Overall, our data suggest that miR-222-3p could alleviate the mitochondrial pathway-mediated apoptosis and motor dysfunction in rats after SCI by targeting Bbc3 and Bim.
Asunto(s)
MicroARNs , Traumatismos de la Médula Espinal , Ratas , Animales , Ratas Sprague-Dawley , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Caspasa 9/farmacología , Proteína X Asociada a bcl-2/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , MicroARNs/metabolismo , Apoptosis , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Médula Espinal/metabolismoRESUMEN
OBJECTIVE: To investigate the effect and mechanism of Pinus massoniana needle extracts (PNE) on oxidative stress injury in cerebral ischemia reperfusion rats. METHODS: The SD male rats were randomly divided into sham group, model control group, Edaravone (3â mg/kg) group, PNE low-dose (200â mg/kg), medium-dose (400â mg/kg) and high-dose (800â mg/kg) groups. PNE was administered by gavage for 7â d before modeling and 6â h after modeling in PNE treatment groups; Edaravone was given by intraperitoneal injection 7â d before modeling and 6â h after reperfusion. The rat model of cerebral ischemia reperfusion injury was established by middle cerebral artery occlusion method. After 24â h of reperfusion, the neurological deficit score, brain water content and cerebral infarction volume of rats were measured. The pathological changes of cerebral cortex and hippocampus were observed by HE staining, and the number of normal nerve cells was counted. The apoptosis rate of neurons in cerebral cortex was detected by TUNEL method. The content of nitric oxide (NO), malondialdehyde (MDA) and superoxide dismutase (SOD) activity in ischemic brain tissue were detected. The protein expression of c-Jun N-terminal kinase (JNK) 3, phosphorylated JNK3 (p-JNK3), B-cell lymphoma protein(Bcl) -2, Bcl-2 associated X (Bax), cytochrome C and caspase-3 in cerebral cortex were detected by Western blotting method. RESULTS: Compared with the model control group, the behavioral score, brain water content and cerebral infarction volume in PNE groups were significantly reduced (all P<0.05), the pathological damage of cerebral cortex and hippocampal CA1 area was significantly alleviated, and the number of normal nerve cells in ischemic cortex and hippocampal CA1 area was increased (all P<0.05). The medium-dose PNE group had the best effect. Compared with the model control group, the apoptosis rate of cortical neurons, the content of NO and MDA in cerebral cortex, the ratio of p-JNK3/JNK3, the expression level of cytochrome C and caspase-3 protein in PNE medium-dose group were significantly reduced , and the activity of SOD, the Bcl-2/Bax ratio were significantly improved (all P<0.05). CONCLUSION: PNE ameliorates brain injury after cerebral ischemia reperfusion in rats, which may be related to scavenging NO and MDA, inhibiting oxidative stress-mediated JNK3/caspase-3 signsal transduction to inhibit neuronal apoptosis.
Asunto(s)
Isquemia Encefálica , Estrés Oxidativo , Extractos Vegetales , Daño por Reperfusión , Animales , Masculino , Ratas , Apoptosis , Proteína X Asociada a bcl-2/metabolismo , Proteína X Asociada a bcl-2/farmacología , Proteína X Asociada a bcl-2/uso terapéutico , Isquemia Encefálica/tratamiento farmacológico , Caspasa 3/metabolismo , Caspasa 3/farmacología , Citocromos c/metabolismo , Citocromos c/farmacología , Citocromos c/uso terapéutico , Edaravona/farmacología , Edaravona/uso terapéutico , Infarto de la Arteria Cerebral Media , Ratas Sprague-Dawley , Reperfusión , Daño por Reperfusión/prevención & control , Daño por Reperfusión/tratamiento farmacológico , Transducción de Señal , Superóxido Dismutasa , Extractos Vegetales/farmacología , Pinus/químicaRESUMEN
Objective: To investigate the effect of arsenic and its main metabolites on the apoptosis of human lung adenocarcinoma cell line A549 and the expression of pro-apoptotic genes Bad and Bik. Methods: In October 2020, A549 cells were recovered and cultured, and the cell viability was detected by the cell counting reagent CCK-8 to determine the concentration and time of sodium arsenite exposure to A549. The study was divided into NaAsO(2) exposure groups and metobol: le expoure groups: the metabolite comparison groups were subdivided into the control group, the monomethylarsinic acid exposure group (60 µmol/L) , and the dimethylarsinic acid exposure group (60 µmol/L) ; sodium arsenite dose groups were subdivided into 4 groups: control group (0) , 20, 40, 60 µmol/L sodium arsenite NaAsO(2). Hoechst 33342/propidium iodide double staining (Ho/PI) was used to observe cell apoptosis and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of Bad and Bik mRNA in cells after exposure. Western blotting was used to detect the protein expressions of Bad, P-Bad-S112, Bik, cleaved Bik and downstream proteins poly ADP-ribose polymerase PARP1 and cytochrome C (Cyt-C) , using spectrophotometry to detect the activity changes of caspase 3, 6, 8, 9. Results: Compared with the control group, the proportion of apoptotic cells in the 20, 40, and 60 µmol/L NaAsO(2) dose groups increased significantly (P<0.01) , and the expression levels of Bad, Bik mRNA, the protein expression levels of Bad, P-Bad-S112, Bik, cleaved Bik, PARP1, Cyt-C were increased (all P<0.05) , and the activities of Caspase 3, 6, 8, and 9 were significantly increased with significantly differences (P<0.05) . Compared with the control group, the expression level of Bad mRNA in the DMA exposure group (1.439±0.173) was increased with a significant difference (P=0.024) , but there was no significant difference in the expression level of Bik mRNA (P=0.788) . There was no significant differences in the expression levels of Bad and Bik mRNA in the poison groups (P=0.085, 0.063) . Compared with the control group, the gray values of proteins Bad, Bik, PARP1 and Cyt-C exposed to MMA were 0.696±0.023, 0.707±0.014, 0.907±0.031, 1.032±0.016, and there was no significant difference between the two groups (P=0.469, 0.669, 0.859, 0.771) ; the gray values of proteins Bad, Bik, PARP1 and Cyt-C exposed to DMA were 0.698±0.030, 0.705±0.022, 0.908±0.015, 1.029±0.010, and there was no difference between the two groups (P=0.479, 0.636, 0.803, 0.984) . Conclusion: Sodium arsenite induces the overexpression of Bad and Bik proteins, initiates the negative feedback regulation of phosphorylated Bad and the degradation of Bik, activates the downstream proteins PARP1, Cyt-C and Caspase pathways, and mediates the apoptosis of A549 cells.
Asunto(s)
Arsénico , Venenos , Células A549 , Adenosina Difosfato Ribosa/farmacología , Apoptosis , Proteínas Reguladoras de la Apoptosis , Arsenitos , Ácido Cacodílico/farmacología , Caspasa 3 , Caspasas/farmacología , Citocromos c/farmacología , Humanos , Proteínas Mitocondriales/farmacología , Propidio/farmacología , ARN Mensajero , Sincalida/farmacología , Compuestos de Sodio , Proteína Letal Asociada a bcl/metabolismoRESUMEN
Furanocoumarins, the secondary metabolites of plants, are considered to be natural insecticides and fungicides because they prevent the invasion of plant pathogenic microorganisms and the predation of herbivorous insects. In this study, novel 2-arylfuranocoumarin derivatives were designed to synthesize by condensation, esterification, bromination, and Wittig reaction. The results showed an excellent photosensitive activity of 2-thiophenylfuranocoumarin (I34). Cell Counting Kit-8 detected that I34 could inhibit the proliferation of Spodoptera frugiperda (Sf9) cells in a time- and concentration-dependent manner under ultraviolet A (UV-A) light for 3 min. The inverted microscope revealed that cells treated with I34 swelled, the membrane was ruptured, and apoptotic bodies appeared. The flow cytometry detected that I34 could induce apoptosis of Sf9 cells, increase the level of intracellular reactive oxygen species (ROS), decrease the mitochondrial membrane potential, and block cell cycle arrest in the G2/M phase. Transmission electron microscopy detected cell mitochondrial cristae damage, matrix degradation, and mitochondrial vacuolation. Further enzyme activity detection revealed that the enzyme activities of apoptosis-related proteins caspase-3 and caspase-9 increased significantly (p < 0.05). Finally, Western blotting analysis detected that the phosphorylation level of Akt and Bad and the expression of the apoptosis inhibitor protein Bcl-XL were inhibited, cleaved-PARP and P53 were increased, and cytochrome C was released from the mitochondria into the cytoplasm. Moreover, under UV-A irradiation, I34 promoted the increase in ROS in Sf9 cells, activated the mitochondrial apoptotic signal transduction pathway, and finally, inhibited cell proliferation. Thus, novel furanocoumarins exhibit a potential application prospect as a biochemical pesticide.
Asunto(s)
Fungicidas Industriales , Furocumarinas , Insecticidas , Plaguicidas , Animales , Caspasa 9/metabolismo , Caspasa 9/farmacología , Spodoptera/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Caspasa 3/metabolismo , Insecticidas/farmacología , Insecticidas/metabolismo , Fungicidas Industriales/farmacología , Inhibidores de Poli(ADP-Ribosa) Polimerasas/metabolismo , Inhibidores de Poli(ADP-Ribosa) Polimerasas/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Mitocondrias , Potencial de la Membrana Mitocondrial , Apoptosis , Proliferación Celular , Furocumarinas/farmacologíaRESUMEN
Aflatoxin B1 (AF) is an unavoidable environmental pollutant that contaminates food, feed, and grains, which seriously threatens human and animal health. Arabic gum (AG) has recently evoked much attention owing to its promising therapeutic potential. Thus, the current study was conducted to look into the possible mechanisms beyond the ameliorative activity of AG against AF-inflicted hepatic injury. Male Wistar rats were assigned into four groups: Control, AG (7.5 g/kg b.w/day, orally), AF (200 µg/kg b.w), and AG plus AF group. AF induced marked liver damage expounded by considerable changes in biochemical profile and histological architecture. The oxidative stress stimulated by AF boosted the production of plasma malondialdehyde (MDA) level along with decreases in the total antioxidant capacity (TAC) level and glutathione peroxidase (GPx) activity. Additionally, AF exposure was associated with down-regulation of the nuclear factor erythroid2-related factor2 (Nrf2) and superoxide dismutase1 (SOD1) protein expression in liver tissue. Apoptotic cascade has also been evoked following AF-exposure, as depicted in overexpression of cytochrome c (Cyto c), cleaved Caspase3 (Cl. Casp3), along with enhanced up-regulation of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin (IL)-6, inducible nitric oxide synthase (iNOS), and nuclear factor kappa-B transcription factor/p65 (NF-κB/p65) mRNA expression levels. Interestingly, the antioxidant and anti-inflammatory contents of AG may reverse the induced oxidative damage, inflammation, and apoptosis in AF-exposed animals.
Asunto(s)
Contaminantes Ambientales , Factor 2 Relacionado con NF-E2 , Aflatoxina B1/metabolismo , Animales , Antiinflamatorios/farmacología , Antioxidantes/metabolismo , Antioxidantes/farmacología , Caspasa 3/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Contaminantes Ambientales/metabolismo , Glutatión Peroxidasa/metabolismo , Mediadores de Inflamación/metabolismo , Interleucinas/metabolismo , Hígado/metabolismo , Masculino , Malondialdehído/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Estrés Oxidativo , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Superóxido Dismutasa-1/metabolismo , Superóxidos/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Nickel nanoparticles are widely used in the industry and may affect the reproductive system. The potential molecular mechanism of exposing the first-trimester trophoblast cell line (HTR-8/SVneo) to nickel nanoparticles remains unclear. Hence, the aim of this study was to investigate the in vitro cytotoxicity of Ni NPs on HTR-8/SVneo cells. HTR-8/SVneo cells were subjected to various concentrations (0, 2.5, 5, 7.5, 10, and 12.5 µg/cm2) of Ni NPs. The toxicity of the Ni NPs was evaluated in HTR-8/SVneo cells by measuring cell viability. The underlying mechanism of nickel nanoparticles toxicity to HTR-8/SVneo cells was determined by measuring the content of intracellular reactive oxygen species, mitochondrial membrane potential, and the rate of cell apoptosis and cell cycle, by measuring adenosine triphosphate levels, intracellular lipid peroxidation malondialdehyde, total superoxide dismutase, and CuZn/Mn-SOD activities, and by determining proteins related to Nrf2, MAPK, and Cytochrome c. Our results showed that the nickel nanoparticles treatment reduced the viability of HTR-8/SVneo cells, while it increased their oxidative stress and lowered their mitochondrial respiratory capacity. Additionally, the nickel nanoparticles treatment induced cell S-phase arrest and apoptosis. These molecular events may be linked to the oxidative stress-Nrf2 pathway/MAPK/Caspase 3 cascade. Thus, nickel nanoparticles exert cytotoxic effects on HTR-8/SVneo cells, which could affect the function of the placenta in human.
Asunto(s)
Nanopartículas , Trofoblastos , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/farmacología , Caspasa 3/metabolismo , Caspasas/metabolismo , Caspasas/farmacología , Citocromos c/metabolismo , Citocromos c/farmacología , Femenino , Humanos , Malondialdehído , Nanopartículas del Metal , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Níquel/metabolismo , Níquel/toxicidad , Embarazo , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismo , Trofoblastos/metabolismoRESUMEN
Acute promyelocytic leukemia (APL) is a specific subtype of acute myelogenous leukemia (AML) characterized by the proliferation of abnormal promyelocytes. Realgar, a Chinese medicine containing arsenic, can be taken orally. Traditional Chinese medicine physicians have employed realgar to treat APL for over a thousand years. Therefore, realgar may be a promising candidate for the treatment of APL. Nevertheless, the underlying mechanism behind realgar therapy is largely unclear. The present study aimed to investigate the effect of realgar on cell death in the APL cell line (NB4) in vitro and to elucidate the underlying mechanism. In this study, after APL cells were treated with different concentrations of realgar, the cell survival rate, apoptotic assay, morphological changes, ATP levels and cell cycle arrest were assessed. The expression of Bcl-2, Bax, Cytochrome C (Cyt-C) and apoptosis-inducing factor (AIF) at the mRNA and protein levels were also measured by immunofluorescence, quantitative PCR (qPCR) and Western blotting. We found that realgar could significantly inhibit APL cell proliferation and cell death in a time- and dose-dependent manner. Realgar effectively decreased the ATP levels in APL cells. Realgar also induced APL cell cycle arrest at the S and G2/M phases. Following realgar treatment, the mRNA and protein levels of Bcl-2 were significantly downregulated, whereas the levels of Bax, Cyt-C, and AIF were significantly upregulated. In summary, realgar can induce APL cell death via the Bcl-2/Bax/Cyt-C/AIF signaling pathway, suggesting that realgar may be an effective therapeutic for APL.
Asunto(s)
Arsénico , Leucemia Promielocítica Aguda , Adenosina Trifosfato , Apoptosis , Factor Inductor de la Apoptosis/metabolismo , Arsénico/metabolismo , Arsénico/farmacología , Arsénico/uso terapéutico , Arsenicales , Muerte Celular , Línea Celular Tumoral , Citocromos c/metabolismo , Citocromos c/farmacología , Citocromos c/uso terapéutico , Humanos , Leucemia Promielocítica Aguda/tratamiento farmacológico , Leucemia Promielocítica Aguda/metabolismo , Medicina Tradicional China , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero , Transducción de Señal , Sulfuros , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
OBJECTIVE: To explore the association between oxidative stress (OS) and Kashin-Beck disease (KBD). METHODS: Terms associated with "KBD" and "OS" were searched in the six different databases up to October 2021. Stata 14.0 was used to pool the means and standard deviations using random-effect or fixed-effect model. The differentially expressed genes in the articular chondrocytes of KBD were identified, the OS related genes were identified by blasting with the GeneCards. The KEGG pathway and gene ontology enrichment analysis was conducted using STRING. RESULTS: The pooled SMD and 95% CI showed hair selenium (-4.59; -6.99, -2.19), blood selenium (-1.65; -2.86, -0.44) and glutathione peroxidases (-4.15; -6.97, -1.33) levels were decreased in KBD, whereas the malondialdehyde (1.12; 0.60, 1.64), nitric oxide (2.29; 1.31, 3.27), nitric oxide synthase (1.07; 0.81, 1.33) and inducible nitric oxide synthase (1.69; 0.62, 2.77) were increased compared with external controls. Meanwhile, hair selenium (-2.71; -5.32, -0.10) and glutathione peroxidases (-1.00; -1.78, -0.22) in KBD were decreased, whereas the malondialdehyde (1.42; 1.04, 1.80), nitric oxide (3.08; 1.93, 4.22) and inducible nitric oxide synthase (0.81; 0.00, 1.61) were elevated compared with internal controls. Enrichment analysis revealed apoptosis was significantly correlated with KBD. The significant biological processes revealed OS induced the release of cytochrome c from mitochondria. The cellular component of OS located in the mitochondrial outer membrane. CONCLUSIONS: The OS levels in KBD were significantly increased because of selenium deficiency, OS mainly occurred in mitochondrial outer membrane, released of cytochrome c from mitochondria, and induced apoptotic signaling pathway.
Asunto(s)
Enfermedad de Kashin-Beck , Selenio , Humanos , Enfermedad de Kashin-Beck/genética , Enfermedad de Kashin-Beck/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Selenio/metabolismo , Biología Computacional , Óxido Nítrico/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Estrés Oxidativo , Malondialdehído/farmacología , Glutatión/metabolismo , Glutatión/farmacología , Peroxidasas/metabolismo , Peroxidasas/farmacologíaRESUMEN
The main objective of this study was to explore the effects of dietary selenium yeast and jujube powder on mitochondrial oxidative damage and cell apoptosis of broilers during postmortem aging, chicken breasts of broilers fed diets supplemented with different concentrations of selenium yeast and jujube powder were used as research subjects. With the prolongation of postmortem aging time, the levels of reactive oxygen species (ROS), carbonyl content, mitochondrial permeability transition pore (MPTP) openness, and mitochondrial membrane permeability increased significantly (P < 0.05). The contents of the sulfhydryl, mitochondrial membrane potential, shear force, and cytochrome C (Cyt-c) reduction level decreased significantly (P < 0.05). The activity of Caspase-3 and Caspase-9 increased from 0 to 24 h postmortem but fell from 24 to 72 h postmortem. Compared with the control group, dietary selenium yeast and jujube powder significantly reduced mitochondrial oxidative damage. They greatly increased the shear force, mitochondrial membrane potential, and Cyt-c reduction levels (P < 0.05). Among them, the combination group of high-dose selenium yeast and jujube powder had more significant effects on ROS scavenging, reducing cell membrane permeability, protecting cell membrane integrity, and increasing Cyt-c reduction level (P < 0.05). In conclusion, cell apoptosis intensifies during the chicken breast's aging time, and muscle tenderness continues. Still, different doses of dietary selenium yeast and jujube powder can inhibit mitochondrial oxidation to various degrees. The combined group of selenium yeast and jujube powder with 0.6 mg·kg-1 has the best effect. This study is of great significance for applying natural antioxidant ingredients such as selenium yeast and jujube powder in the development and utilization of poultry feed.
Asunto(s)
Selenio , Ziziphus , Animales , Antioxidantes/metabolismo , Apoptosis , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Caspasa 9/farmacología , Pollos , Citocromos c/metabolismo , Citocromos c/farmacología , Suplementos Dietéticos , Poro de Transición de la Permeabilidad Mitocondrial , Estrés Oxidativo , Polvos/farmacología , Especies Reactivas de Oxígeno/metabolismo , Saccharomyces cerevisiae/metabolismo , Selenio/metabolismo , Selenio/farmacología , Ziziphus/metabolismoRESUMEN
Di-(2-ethylhexyl) phthalate (DEHP) is one of the most used plasticizers which have contaminated environment widely, and its extensive use causes female reproductive injury. Melatonin has a substantial protective effect against female reproductive toxicity. This study was undertaken to investigate the influence of melatonin on DEHP-induced damage of human granulosa cells (GCs) in vitro and explore the potential mechanisms. Here, we found that melatonin treatment alleviated DEHP-induced human GCs apoptosis and improved mitochondrial function via inhibiting dynamin-related protein 1 (Drp1) mediated mitochondrial fission. Melatonin inhibited the expression, activation and oligomerization of Drp1, which decreased translocation of Drp1 to mitochondria in DEHP-exposed human GCs. Inhibition of mitochondrial fission reduced intracellular reactive oxygen species (ROS) production, sustained mitochondrial membrane potential and decreased cytochrome c release. Further research showed that AMPK-PGC-1α signal pathway was involved in the inhibition of melatonin on Drp1 expression and activation. Melatonin treatment promoted AMPK activation suppressed by DEHP, and activated AMPK recovered the balance of Drp1 phosphorylation at Ser616 and Ser637 sites and enhanced PGC-1α expression. Moreover, PGC-1α could prevent mitochondrial fission by decreasing Drp1 expression directly via binding to its promoter. In contrast, blocking of AMPK or PGC-1α with specific inhibitor negated the protective effects of melatonin on mitochondrial homeostasis and GCs apoptosis. In summary, our results indicated the protective effects of melatonin on improving mitochondrial function and attenuating cells injury in DEHP-exposed human GCs. Melatonin treatment may be a promising therapeutic approach against DEHP-induced reproductive disorder.
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Dietilhexil Ftalato , Melatonina , Proteínas Quinasas Activadas por AMP/metabolismo , Apoptosis , Citocromos c/metabolismo , Citocromos c/farmacología , Dietilhexil Ftalato/toxicidad , Dinaminas/metabolismo , Dinaminas/farmacología , Femenino , Células de la Granulosa/metabolismo , Humanos , Melatonina/farmacología , Dinámicas Mitocondriales , Ácidos Ftálicos , Plastificantes/toxicidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Plant diseases cause a huge impact on food security and are of global concern. While application of agrochemicals is a common approach in the control of plant diseases currently, growing drug resistance and the impact of off-target effects of these compounds pose major challenges. The identification of pathogenicity-related virulence mechanisms and development of new chemicals that target these processes are urgently needed. One such virulence mechanism is the detoxification of reactive oxygen species (ROS) generated by host plants upon attack by pathogens. The machinery of ROS detoxification might therefore serve as a drug target for preventing plant diseases, but few anti-ROS-scavenging drugs have been developed. Here, we show that in the model system Botrytis cinerea secretion of the cytochrome c-peroxidase, BcCcp1 removes plant-produced H2O2 and promotes pathogen invasion. The peroxidase secretion is modulated by a Tom1-like protein, BcTol1, through physical interaction. We show that BcTol1 is regulated at different levels to enhance the secretion of BcCcp1 during the early infection stage. Inactivation of either BcTol1 or BcCcp1 leads to dramatically reduced virulence of B. cinerea. We identify two BcTol1-targeting small molecules that not only prevent B. cinerea invasion but also have effective activity against a wide range of plant fungal pathogens without detectable effect on the hosts. These findings reveal a conserved mechanism of ROS detoxification in fungi and provide a class of potential fungicides to control diverse plant diseases. The approach described here has wide implications for further drug discovery in related fields.
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Fungicidas Industriales , Botrytis/metabolismo , Citocromos c/metabolismo , Citocromos c/farmacología , Proteínas Fúngicas/metabolismo , Fungicidas Industriales/metabolismo , Fungicidas Industriales/farmacología , Peróxido de Hidrógeno/farmacología , Peroxidasas/metabolismo , Peroxidasas/farmacología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Condensed tannins the polyphenolic compounds that are widespread in plants have been proved to have antitumor potential. Here, we purified the bioactive condensed tannins from leaves of Ulmus pumila L. and explored their structural characteristics, antitumor effect on TFK-1 cholangiocarcinoma cells as well as the related potential mechanism. The UV-Vis, FT-IR spectroscopy, ESI-Full-MS, and thiolysis-HPLC-ESI-MS demonstrated that U. pumila condensed tannins (UCTs) consisted essentially of procyanidins with epicatechin as the main flavan-3-ol extension unit. The UCTs could significantly reduce the survival rate of human cholangiocarcinoma TFK-1, SK-CHA-1, and MZ-CHA-1 cells with the better inhibitory effect on TFK-1 cell proliferation. Flow cytometric assay showed that UCTs affected TFK-1 survival by G2/M phase arrest and inducing apoptosis in a dose-dependent manner. In addition, a total of 6592 differentially expressed genes (DEGs), consisting of 94 upregulated and 6498 downregulated DEGs, were identified between untreated and UCTs-treated TFK-1 cells using RNA-seq technology. Enrichment analysis based on the KEGG database revealed that these DEGs were closely associated with cell cycle and p53 apoptotic signaling pathways. Furthermore, qRT-PCR confirmed that treatment of UCTs to TFK-1 cells caused significant changes in the expression of cyclin E, cdc25 A, cytochrome c, caspase-3, and caspase-8. These results indicated that UCTs exhibited the growth inhibition effect on TFK-1 cells possibly via G2/M cell cycle arrest and activation of caspase-cascade to induce apoptosis, and had potential as an anti-cholangiocarcinoma drug for further development. PRACTICAL APPLICATIONS: Ulmus pumila L. as a valuable tree species has been widely used in fields of medicine and food. Condensed tannins, the polyphenolic compounds widespread in plants, have been proved to have antitumor potential and be safe to normal cells. In this study, the condensed tannins from leaves of U. pumila (UCTs) remarkably suppressed cholangiocarcinoma (CCA) cell viability possibly via G2/M cell cycle arrest and activation of caspase-cascade to induce apoptosis. The results provided evidence for the application of UCTs as a potential therapeutic drug for CCA tumor.
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Neoplasias de los Conductos Biliares , Catequina , Colangiocarcinoma , Proantocianidinas , Ulmus , Apoptosis , Neoplasias de los Conductos Biliares/tratamiento farmacológico , Neoplasias de los Conductos Biliares/metabolismo , Neoplasias de los Conductos Biliares/patología , Conductos Biliares Intrahepáticos/metabolismo , Conductos Biliares Intrahepáticos/patología , Caspasa 3/metabolismo , Caspasa 8/genética , Caspasa 8/metabolismo , Caspasa 8/farmacología , Caspasas/metabolismo , Caspasas/farmacología , Caspasas/uso terapéutico , Catequina/farmacología , Puntos de Control del Ciclo Celular , División Celular , Colangiocarcinoma/tratamiento farmacológico , Colangiocarcinoma/metabolismo , Colangiocarcinoma/patología , Ciclina E/metabolismo , Ciclina E/farmacología , Citocromos c/metabolismo , Citocromos c/farmacología , Citocromos c/uso terapéutico , Humanos , Proantocianidinas/farmacología , Espectroscopía Infrarroja por Transformada de Fourier , Proteína p53 Supresora de Tumor , Ulmus/metabolismoRESUMEN
This work aimed for the first time to provide detailed insights into thymol and trans-cinnamaldehyde's mechanisms of action on the food-spoilage yeast Zygosaccharomyces rouxii and offers evidence in favor of the activation of an apoptosis-like phenotype. The action mechanisms of thymol and trans-cinnamaldehyde were investigated by the measurement of a series of typical apoptotic features using flow cytometer or microplate reader. Moreover, quantitative reverse transcription PCR (QRT-PCR) was performed to investigate the effects of thymol and trans-cinnamaldehyde on the transcription of key regulators of apoptosis in Z. rouxii. The results indicated that the treatment of Z. rouxii with thymol or trans-cinnamaldehyde (minimum inhibitory and subinhibitory concentrations) triggered reactive oxygen species (ROS) accumulation, elevated intracellular Ca2+ level, and depolarized mitochondrial membrane potential (MMP) coupled with hallmarks of apoptosis including mitochondrial cytochrome c (cyt c) release, metacaspase activation, phosphatidylserine (PS) exposure, and DNA fragmentation. Moreover, thymol or trans-cinnamaldehyde treatment upregulated the transcription of proapoptotic regulators including Yca1, Dnm1, Nuc1, Ndi1, and Mmi1 and downregulated the transcription of antiapoptotic regulators of Fis1 and Cdc48, further confirming the apoptosis induced by thymol and trans-cinnamaldehyde in Z. rouxii. In summary, thymol and trans-cinnamaldehyde probably induced apoptosis through a metacaspase-dependent mitochondrial pathway in Z. roxuii. These findings provide theoretical support for the development of safe and efficient agents used in food preservation. PRACTICAL APPLICATION: The results will provide a new idea for the systematic analysis of the antifungal mechanisms of thymol and trans-cinnamaldehyde and also provide a theoretical support for the development and application of natural food preservatives, which is of positive significance for the effective control of the spoilage caused by Z. rouxii in food processing and storage and the protection of consumers' health.
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Proteínas de Saccharomyces cerevisiae , Zygosaccharomyces , Acroleína/análogos & derivados , Antifúngicos/farmacología , Apoptosis , Citocromos c/farmacología , Complejo I de Transporte de Electrón , Endonucleasas , Exonucleasas , Conservantes de Alimentos/farmacología , Proteínas Mitocondriales/farmacología , Fosfatidilserinas/farmacología , Especies Reactivas de Oxígeno , Saccharomyces cerevisiae/genética , Saccharomycetales , Timol/farmacología , Zygosaccharomyces/genéticaRESUMEN
Vaginal candidiasis is a medical condition characterized by the overgrowth of Candida spp. in the vaginal cavity with complex recurrent pathogenicity as well as tolerance to antifungal therapy and hence is awaiting more safe and effective treatments. This work aimed to assess the potential antifungal activity of galloylquinic acid compounds (GQAs) from Copaifera lucens leaves against vaginal Candida albicans. The antifungal susceptibility test was performed against 20 isolates of multidrug-resistant (MDR) C. albicans using agar diffusion and broth microdilution assays. The results showed that GQAs exhibited strong antagonistic activity against the test isolates, with inhibition zone diameters ranging from 26 to 38 mm and low MICs (1 to 16 µg/mL) as well as minimum fungicidal concentrations (2 to 32 µg/mL). The MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay confirmed the safety of GQAs against the Vero cell line, showing a 50% inhibitory concentration (IC50) of 168.17 mg/mL. A marked difference in the growth pattern of the treated and untreated pathogens was also observed, where a concentration-dependent reduction in the growth rate occurred. Moreover, a pronounced fungicidal effect was demonstrated 6 h after treatment with 1× the minimum fungicidal concentration (MFC), as evidenced by time-kill assays, where the number of survivors was decreased a 6-fold. GQAs effectively inhibited and eradicated about 80% of C. albicans biofilm at 6 µg/mL and 32 µg/mL, respectively. Interestingly, GQAs disturbed the fungal membrane integrity, induced cell lysis, and reduced the virulence factors (proteinase and phospholipase) as well as the catalase activity. Moreover, the ergosterol content in the plasma membrane decreased in a concentration-dependent manner. Additionally, the altered mitochondrial membrane potential was associated with an increased release of cytochrome c from mitochondria to the cytosol, suggesting the initiation of early apoptosis in GQA-treated cells. Transcriptional analysis revealed that all test genes encoding virulence traits, including SAP1, PLB1, LIP1, HWP1, and ALS1, were markedly downregulated in GQA-treated cells compared to the control. The in vivo murine model of vaginal candidiasis further confirmed the therapeutic activity of GQAs (4 mg/kg of body weight) against C. albicans. This work comprehensively evaluated the antifungal, antivirulence, and antibiofilm activities of GQAs against C. albicans isolates using in vitro and in vivo models, providing molecular-level insights into the antifungal mechanism of action and experimental evidence that supports the potential use of GQAs for the treatment of vaginal candidiasis. IMPORTANCE Our work presents a new perspective on the potential use of GQAs as safe and highly effective phytochemicals against MDR C. albicans. This microorganism colonizes the human vaginal epithelium, causing vaginal candidiasis, a condition characterized by recurrent pathogenicity and tolerance to traditional antifungal therapy. Based on the results of in vitro tests, our study reports GQAs antifungal modes of action. These compounds exhibited an anticandidal effect by deactivating the fungal hydrolytic enzymes, reducing ergosterol content in the plasma membrane, altering the potential of the mitochondrial membrane, and inducing apoptosis. Additionally, GQAs showed high activity in eradicating the biofilm formed by the fungus via the downregulation of HWP1, ALS, SAP, PLB, and LIP genes, which are constitutively expressed in the biofilm. In an in vivo murine model of vaginal candidiasis, GQAs further demonstrated strong evidence of their effectiveness as an antifungal therapy. In this regard, our findings provide novel insights into the potential therapeutic use of these phytoactive molecules for vaginal candidiasis treatment.
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Candidiasis Vulvovaginal , Candidiasis , Fabaceae , Femenino , Ratones , Humanos , Animales , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Modelos Animales de Enfermedad , Citocromos c/farmacología , Citocromos c/uso terapéutico , Agar/farmacología , Agar/uso terapéutico , Catalasa/farmacología , Catalasa/uso terapéutico , Candidiasis Vulvovaginal/tratamiento farmacológico , Candidiasis Vulvovaginal/microbiología , Candida albicans , Candidiasis/tratamiento farmacológico , Biopelículas , Pruebas de Sensibilidad Microbiana , Factores de Virulencia , Ergosterol , Fosfolipasas/farmacología , Fosfolipasas/uso terapéutico , Péptido Hidrolasas/farmacología , Péptido Hidrolasas/uso terapéuticoRESUMEN
Taurine plays an important role in neural growth and function from early to adult life, particularly in learning and memory via BDNF action. This study tested the hypothesis that BDNF differentially potentiates entorhinal-hippocampal synaptic transmission in vivo in adult rats. In anesthetized male Sprague-Dawley rats, a stainless steel recording electrode with an attached microinjector was placed into CA1 and the dentate gyrus to record fEPSP, and a paired stainless steel electrode was inserted into entorhinal cortex for continuous paired-pulse stimulation of that brain region. In the dentate gyrus, microinjection of BDNF resulted in a gradual increase in the peak slope of the fEPSP. Following the infusion, the peak fEPSP began to rise in about 8 min, reached a maximum of 120 ± 2% (from baseline) by about 20 min, and remained near peak elevation (~115%) for more than 30 min. In contrast, the same dose of BDNF when injected into CA1 had no consistent effect on fEPSP slopes in the CA1. Further, an equimolar cytochrome C (horse heart) infusion had no significant effect on fEPSP slopes in either the dentate gyrus or CA1. The potentiation effect of BDNF in the dentate gyrus is consistent with a significant increase in power spectral density of dentate gyrus field potentials at 70-200 Hz, but not at frequencies below 70 Hz. In addition, the CA1 power spectral density was not affected by BDNF (compared to cytochrome C). These data indicate that in vivo BDNF potentiates entorhinal-hippocampal synaptic transmission in dentate gyrus, but not in CA1.