RESUMEN
Smokeless tobacco (SLT) or chewing tobacco has been a highly addictive practice in India across ages, posing major threat to the systemic health and possibly neurodegeneration. Earlier studies showed components of SLT could be harmful to neuronal health. However, mechanism of SLT in neurodegeneration remained unexplored. This study investigated the detrimental role of SLT on differentiated neuronal cell lines, PC12 and SH-SY5Y by using graded doses of water soluble lyophilised SLT. Reduced cell viability, compromised mitochondrial structure and functions were observed when neuronal cell lines were treated with SLT (6 mg/mL) for 24 h. There was reduction of oxidative phosphorylation and aerobic glycolysis as determined by diminution of ATP production (2.5X) and basal respiration (1.9X). Mitochondrial membrane potential was dropped by 3.5 times. Bid, a pro-apoptotic Bcl-2 family protein, has imperative role in regulating mitochondrial outer membrane permeabilization and subsequent cytochrome c release leading to apoptosis. This article for the first time indicated the involvement of Bid in SLT mediated neurotoxicity and possibly neurodegeneration. SLT treatment enhanced expression of cleaved-Bid in time dependent manner. The involvement of Bid was further confirmed by using Bid specific shRNA which reversed the effects of SLT and conferred significant protection from apoptosis up to 72 h. Thus, our results clearly indicated that SLT induced neuronal cell death occurred via production of ROS, alteration of mitochondrial morphology, membrane potential and oxidative phosphorylation, inactivation of survival pathway and activation of apoptotic markers mediated by Bid. Therefore, Bid could be a potential future therapeutic target for SLT induced neurodegeneration.
Asunto(s)
Neuronas/patología , Tabaco sin Humo/toxicidad , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Línea Celular Tumoral , Citocromos c/metabolismo , Daño del ADN , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fosforilación Oxidativa , Células PC12 , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Especies Reactivas de Oxígeno/metabolismo , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
AIM: The present work provides first-time empirical and molecular interaction evidence to establish the higher biofunctionality of a therapeutic lipid, α-eleostearic acid (ESA), encapsulated in a novel and thoroughly characterized biocompatible nanoemulsion (NE) system (particle size <200 nm). MATERIALS & METHODS: A novel methodology was employed to fabricate novel formulations of ESA. Molecular biological tools and assays were used to arrive at definite conclusions. RESULTS: The proinflammatory profile was found to be significantly mitigated in the hypersensitized rats administered with the ESA-NE formulation more emphatically as compared with ESA-conventional emulsion in both in vivo and ex vivo models. CONCLUSION: The novel ESA-NE formulation shows a lot of palpable promise for clinical applications.
Asunto(s)
Emulsiones/química , Inflamación/patología , Ácidos Linolénicos/química , Animales , Ciclo Celular/efectos de los fármacos , Células Cultivadas , Emulsiones/uso terapéutico , Citometría de Flujo , Voluntarios Sanos , Humanos , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Ácidos Linolénicos/uso terapéutico , Masculino , Óxido Nítrico/metabolismo , Tamaño de la Partícula , Peroxidasa/metabolismo , Aceites de Plantas/química , RatasRESUMEN
The current investigation has identified the biomarkers associated with severity of disability and correlation among plethora of systemic, cellular and molecular parameters of intellectual disability (ID) in a rehabilitation home. The background of study lies with the recent clinical evidences which identified complications in ID. Various indicators from blood and peripheral system serve as potential surrogates for disability related changes in brain functions. ID subjects (Male, age 10 ± 5 yrs, N = 45) were classified as mild, moderate and severe according to the severity of disability using standard psychometric analysis. Clinical parameters including stress biomarkers, neurotransmitters, RBC morphology, expressions of inflammatory proteins and neurotrophic factor were estimated from PBMC, RBC and serum. The lipid peroxidation of PBMC and RBC membranes, levels of serum glutamate, serotonin, homocysteine, ROS, lactate and LDH-A expression increased significantly with severity of ID whereas changes in RBC membrane ß-actin, serum BDNF, TNF-α and IL-6 was found non-significant. Structural abnormalities of RBC were more in severely disabled children compared to mildly affected ones. The oxidative stress remained a crucial factor with severity of disability. This is confirmed not only by RBC alterations but also with other cellular dysregulations. The present article extends unique insights of how severity of disability is correlated with various clinical, cellular and molecular markers of blood. This unique study primarily focuses on the strong predictors of severity of disability and their associations via brain-blood axis.
Asunto(s)
Biomarcadores/sangre , Niños con Discapacidad/rehabilitación , Eritrocitos/patología , Discapacidad Intelectual/diagnóstico , Adolescente , Niño , Preescolar , Humanos , India , Discapacidad Intelectual/sangre , Discapacidad Intelectual/patología , Peroxidación de Lípido , Masculino , Índice de Severidad de la EnfermedadRESUMEN
Radiation-induced gastrointestinal syndrome occurs due to the clonogenic loss of crypt cells and villi depopulation, resulting in disruption of the mucosal barrier, bacterial invasion, inflammation, and sepsis. In this study, we investigated the role of ferulic acid (FA) against ionising radiation-induced duodenal injury and subsequent alterations in redox signalling events in wild type male Swiss albino mice. Mice were administered with FA at a dose of 50 mg/kg body weight for 5 consecutive days prior to exposure of 2.5, 5 and 10 Gy doses of γ-radiation. Histopathological and electron microscopic images revealed marked duodenal injuries in a dose-dependent manner. FA prevented radiation induced damage and loss of cryptic stem cells and the shortening of duodenal villus length. FA pretreatment further suppressed NF-κB-dependent activation of inflammatory pathways and augmented Nrf2 nuclear translocation with higher expression of Mn-SOD and heme-oxygenase one (HO1) activity to combat with radiation induced duodenal stress. The colocalisation of NF-κB and Nrf2 transcription factors in the nuclei of the duodenum indicated their interaction in radiation and the FA combination group. Moreover, FA treatment inhibited phosphatidyl serine (PS) externalisation, and loss of mitochondrial membrane potential in duodenal cells. Animals exposed to 10-Gy irradiation exhibited over activation of p53, p21, caspase 3, poly ADP ribose polymerase (PARP) and DNA double-strand break which were ameliorated by FA treatment. Therefore, this article first uncovers the modulatory effect of FA on radiation-induced ROS/NF-κB/Nrf2/p53-caspase 3-PARP axis in the duodenum and establishing biological function of FA in protecting duodenum from radiation damage with a detailed mechanistic approach.
Asunto(s)
Ácidos Cumáricos/metabolismo , Duodeno/efectos de la radiación , Rayos gamma/efectos adversos , Irradiación Corporal Total/métodos , Animales , Duodeno/patología , Masculino , Ratones , Especies Reactivas de Oxígeno , Transducción de SeñalRESUMEN
The present study was aimed to evaluate the radioprotective effect of ferulic acid (FA), a naturally occurring plant flavonoid in terms of DNA damage and damage related alterations of repair pathways by gamma radiation. FA was administered at a dose of 50 mg/kg body weight for five consecutive days prior to exposing the swiss albino mice to a single dose of 10 Gy gamma radiation. Ionising radiation induces oxidative damage manifested by decreased expression of Cu, Zn-SOD (SOD stands for super oxide dismutase), Mn-SOD and catalase. Gamma radiation promulgated reactive oxygen species (ROS) mediated DNA damage and modified repair pathways. ROS enhanced nuclear translocation of p53, activated ATM (ataxia telangiectasia-mutated protein), increased expression of GADD45a (growth arrest and DNA-damage-inducible protein) gene and inactivated Non homologous end joining (NHEJ) repair pathway. The comet formation in irradiated mice peripheral blood mononuclear cells (PBMC) reiterated the DNA damage in IR exposed groups. FA pretreatment significantly prevented the comet formation and regulated the nuclear translocation of p53, inhibited ATM activation and expression of GADD45a gene. FA promoted the nuclear translocation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and activated NHEJ repair pathway to overcome ROS mediated oxidative stress and DNA damage. Therefore, the current study stated that FA can challenge the oxidative stress by (i) inducing nuclear translocation of Nrf2, (ii) scavenging ROS, and (iii) activating NHEJ DNA repair process.
Asunto(s)
Ácidos Cumáricos/uso terapéutico , Depuradores de Radicales Libres/uso terapéutico , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo/efectos de los fármacos , Protectores contra Radiación/uso terapéutico , Animales , Compuestos de Bifenilo/química , Catalasa/metabolismo , Resinas Compuestas , Ácidos Cumáricos/química , Ácidos Cumáricos/farmacología , Daño del ADN , Reparación del ADN por Unión de Extremidades , Evaluación Preclínica de Medicamentos , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacología , Rayos gamma , Masculino , Ratones , Oxidación-Reducción , Picratos/química , Plásmidos/química , Protectores contra Radiación/química , Protectores contra Radiación/farmacología , Transducción de Señal , Superóxido Dismutasa/metabolismo , Activación Transcripcional/efectos de los fármacosRESUMEN
PURPOSE: The spleen is a crucial organ manifesting immune functions. Thus, radiation-induced oxidative challenge is vulnerable for the spleen. Our major objective was to protect the spleen from radiation-induced anomalous situations and to identify the signaling pathways involved. MATERIALS AND METHODS: Swiss albino mice were treated with ferulic acid (FA) once in a day at a dose of 50 mg/kg body weight for 5 consecutive days before exposing them to single dose of 10 Gy irradiation. The ROS generation and MMP change were determined by flow cytometry. The expression of different signaling proteins was investigated by immunoblotting and immunocytochemistry. RESULTS: FA pretreatment significantly prevented radiation-induced oxidative stress by downregulating TBARS formation and by upregulating SOD and catalase activity. FA scavenged ROS, prevented the alteration of MMP and downregulated the expression of stress marker Cdc42 and apoptotic markers p53, p21, Bax and PTEN. Cell cycle analysis showed DNA damage induced arrest of cells at subG0/G1 phase. Moreover, pretreatment with FA augmented Bcl2 expression and also increased the level of p-PI3K. CONCLUSION: FA prevented the activation of apoptotic signaling events in the spleen by interfering with the free radical chain reaction and by scavenging superfluous ROS. This is perhaps the first comprehensive study with a mechanistic viewpoint that FA can protect the spleen from ionizing radiation.
Asunto(s)
Traumatismos por Radiación/inmunología , Traumatismos por Radiación/prevención & control , Especies Reactivas de Oxígeno/inmunología , Bazo/efectos de la radiación , Enfermedades del Bazo/inmunología , Enfermedades del Bazo/prevención & control , Animales , Ácidos Cumáricos/administración & dosificación , Citocinas/inmunología , Relación Dosis-Respuesta a Droga , Depuradores de Radicales Libres/administración & dosificación , Masculino , Ratones , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/inmunología , Estrés Oxidativo/efectos de la radiación , Protectores contra Radiación/administración & dosificación , Resultado del Tratamiento , Irradiación Corporal Total/efectos adversosRESUMEN
Smokeless tobacco (SLT) remains a threat amongst a large population across the globe and particularly in India. The oral use of tobacco has been implicated to cause physiological stress leading to extreme toxicological challenge. The study included 47 SLT-users and 44 non-users providing a spectrum of pathophysiological, clinico-biochemical, antioxidant parameters, cell cycle progression study of PBMC and morphological changes of red blood cells (RBC). The expressions of p53, p21, Bax, Bcl-2, IL-6, TNF- α, Cox-2, iNOS were analyzed from thirteen representative SLT-users and twelve non-users. Difference in CRP, random glucose, serum cholesterol, TG, HLDL-C, LDL-C, VLDL-C, neutrophil count, monocyte count, ESR, SOD (PBMC) and TBARS (RBC membrane) were found to be statistically significant (p < 0.05) between the studied groups. The current study confers crucial insight into SLT mediated effects on systemic toxicity and stress. This has challenged the metabolic condition leading to a rise in the inflammatory status, increased apoptosis and RBC membrane damage. The above findings were substantiated with metabolic, clinical and biochemical parameters. This is possibly the first ever in-depth report and remains an invaluable document on the fatal effects of SLT.
Asunto(s)
Apoptosis/efectos de los fármacos , Eritrocitos/metabolismo , Leucocitos Mononucleares/metabolismo , Estrés Fisiológico/efectos de los fármacos , Tabaco sin Humo/efectos adversos , Adulto , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Ciclooxigenasa 2/metabolismo , Eritrocitos/patología , Femenino , Humanos , India , Interleucina-6/metabolismo , Leucocitos Mononucleares/patología , Lípidos/sangre , Masculino , Óxido Nítrico Sintasa de Tipo II/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Radioprotective action of gossypetin (GTIN) against gamma (γ)-radiation-induced oxidative stress in liver was explored in the present article. Our main aim was to evaluate the protective efficacy of GTIN against radiation-induced alteration of liver in murine system. To evaluate the effect of GTIN, it was orally administered to mice at a dose of 30 mg/kg body weight for three consecutive days prior to γ-radiation at a dose of 5 Gy. Radioprotective efficacy of GTIN were evaluated at physiological, cellular, and molecular level using biochemical analysis, comet assay, flow cytometry, histopathology, immunofluorescence, and immunoblotting techniques. Ionizing radiation was responsible for augmentation of hepatic oxidative stress in terms of lipid peroxidation and depletion of endogenous antioxidant enzymes. Immunoblotting and immunofluorescence studies showed that irradiation enhanced the nuclear translocation of nuclear factor kappa B (NF-κB) level, which leads to hepatic inflammation. To investigate further, we found that radiation induced the activation of stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK)-mediated apoptotic pathway and deactivation of the NF-E2-related factor 2 (Nrf2)-mediated redox signaling pathway, whereas GTIN pretreatment ameliorated these radiation-mediated effects. This is the novel report where GTIN rationally validated the molecular mechanism in terms of the modulation of cellular signaling system' instead of ' This is the novel report where GTIN is rationally validated in molecular terms to establish it as promising radioprotective agents. This might be fruitful especially for nuclear workers and defense personnel assuming the possibility of radiation exposure.