Sanjit Dey

Pomegranate reverses methotrexate-induced oxidative stress and apoptosis in hepatocytes by modulating Nrf2-NF-κB pathways

The clinical efficacy of the widely used chemotherapeutic drug methotrexate (MTX) is limited due to its associated hepatotoxicity. Pomegranate polyphenols are of huge health benefits and known to possess remarkable antioxidant properties capable of protecting normal cells from various stimuli-induced oxidative stress and cell death. In this study, we explored the protective role of pomegranate fruit extract (PFE) in ameliorating MTX-induced hepatic damage. Male Swiss albino mice exposed to MTX (20 mg/kg body weight) exhibited distinct markers of toxicity such as increased activities of enzymes alanine transaminase, aspartate transaminase, lactate dehydrogenase and alkaline phosphatase and also increased oxidative stress in liver evidenced by increased ROS generation and lipid peroxidation. Decrease in reduced glutathione levels, superoxide dismutase, catalase, hepatic heme oxygenase 1 and NQO-1 activities were also observed. Tracing the signal transduction pathways, it was seen that MTX exposure significantly increased nuclear translocation of NF-κB coupled with increase in phosphorylated Iκ-B and down-regulation of NF-kappaB-dependent antiapoptotic protein Bcl-2. Treatment with MTX increased the expression of the apoptotic enhancer Rho/Cdc42 as well as the phosphorylation of SAPK/JNK. A shift in the Bax/Bcl-2 ratio towards apoptosis and increase in the caspase 3 level was also evident. Administration of PFE for 7 consecutive days before and after MTX challenge suppressed MTX-induced cell death, mitigated the injurious effects of MTX and offered protection against apoptosis. PFE was shown to reduce ROS generation in hepatocytes by activating the Nrf2-ARE pathway and inhibiting NF-κB as a consequence of which the antioxidant defense mechanism in the liver was up-regulated, thereby conferring protection against MTX-induced hepatotoxicity and apoptosis.

Promising role of ferulic acid, atorvastatin and their combination in ameliorating high fat diet-induced stress in mice.

AIMS The present study evaluated a comparative and combined hepatoprotective effect of atorvastatin (AS) and ferulic acid (F) against high fat diet (HFD) induced oxidative stress in terms of hyperlipidemia, anti-oxidative status, lipid peroxidation and inflammation. MAIN METHODS Male Swiss albino mice were given a diet containing high fat (H) (23.9% wt/wt), supplemented with AS (10mg/kg) or F (100mg/kg) and both (10 and 100mg/kg) for 8weeks. The control mice (C) were fed with normal diet. KEY FINDINGS The H mice exhibited increased body weight; hyperlipidemia; serum level of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6); hepatic lipid profile; lipid accumulation; reactive oxygen species (ROS) of hepatocytes, lipid peroxidation and liver antioxidant capacity was decreased. Immunofluorescent and Western blot assay revealed activation of nuclear factor kappa B (NF-κB) signaling pathway. The addition of F or AS and both in the diet significantly counteracted HFD induced body weight gain; hyperlipidemia; TNF-α, IL-6; hepatic lipid profile; fatty infiltration; NF-κB signaling pathway; ROS; lipid peroxidation and moreover elevated levels of hepatic antioxidant enzymes activity were observed. SIGNIFICANCE Simultaneous treatment with AS, F and their combination protected against HFD induced weight gain and oxidative stress. The protection may be attributed to the hypolipidemic and free radical scavenging activity of AS or F and their combination. This study illustrates that AS and F have relatively similar hypolipidemic, antioxidative, anti-inflammatory actions and the AS+F combination along with HFD has shown outstanding effects as compared to other treated groups.

Role of Ferulic Acid in the Amelioration of Ionizing Radiation Induced Inflammation: A Murine Model

Ionizing radiation is responsible for oxidative stress by generating reactive oxygen species (ROS), which alters the cellular redox potential. This change activates several redox sensitive enzymes which are crucial in activating signaling pathways at molecular level and can lead to oxidative stress induced inflammation. Therefore, the present study was intended to assess the anti-inflammatory role of ferulic acid (FA), a plant flavonoid, against radiation-induced oxidative stress with a novel mechanistic viewpoint. FA was administered (50 mg/kg body wt) to Swiss albino mice for five consecutive days prior to exposing them to a single dose of 10 Gy 60Co γ-irradiation. The dose of FA was optimized from the survival experiment and 50 mg/kg body wt dose showed optimum effect. FA significantly ameliorated the radiation induced inflammatory response such as phosphorylation of IKKα/β and IκBα and consequent nuclear translocation of nuclear factor kappa B (NF-κB). FA also prevented the increase of cycloxygenase-2 (Cox-2) protein, inducible nitric oxide synthase-2 (iNOS-2) gene expression, lipid peroxidation in liver and the increase of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in serum. It was observed that exposure to radiation results in decreased activity of superoxide dismutase (SOD), catalase (CAT) and the pool of reduced glutathione (GSH) content. However, FA treatment prior to irradiation increased the activities of the same endogenous antioxidants. Thus, pretreatment with FA offers protection against gamma radiation induced inflammation.

High prevalence and low awareness, treatment and control of hypertension in Asian Indian women

Hypertension is an important public health problem in India. To determine its prevalence, awareness, treatment and control among women, we performed a nationwide study. Population-based studies among women aged 35–70 years were performed in four urban and five rural locations. Stratified sampling was performed and we enrolled 4608 (rural 2604 and urban 2004) of the targeted 8000 (57%). Demographic details, medical history, diet, physical activity, anthropometry and blood pressure (BP) were recorded. Descriptive statistics are reported. Logistic regression was performed to determine the association of hypertension and its awareness, treatment and control with socioeconomic factors. Age-adjusted prevalence of hypertension (known or BP⩾140/⩾90 mm Hg) was observed in 1672 women (39.2%) (rural 746, 31.5%; urban 926, 48.2%). Significant determinants of hypertension were urban location, greater literacy, high dietary fat, low fibre intake, obesity and truncal obesity (P<0.01). Hypertension awareness was noted in 727 women (42.8%), more in urban (529, 56.8%) than in rural (198, 24.6%). Of these, 38.6% of the women were on treatment (urban 35.7, rural 46.5) and of those treated, controlled blood pressure (<140 and <90 mm Hg) was observed in 21.5% (urban 28.3 vs 10.2). Among hypertensive subjects, treatment was noted in 18.3% (rural 13.1, urban 22.5) and control in 3.9% (rural 1.3, urban 5.9). A significant determinant of low awareness, treatment and control was rural location (multivariate-adjusted P<0.05). There is a high prevalence of hypertension in middle-aged Asian Indian women. Very low awareness, treatment and control status are observed.

Determinants of urban-rural differences in cardiovascular risk factors in middle-aged women in India: a cross-sectional study.

OBJECTIVES Cardiovascular diseases (CVD) are the most important cause of death amongst middle-aged Indian women. To determine prevalence of CVD risk factors and their determinants we performed a nationwide study. METHODS Population based studies amongst women 35-70 years were performed in four urban and five rural locations in India. Location based stratified sampling was performed and we enrolled 4624 (rural 2616, urban 2008) of eligible 8000 women (58%). Demographic details, medical history, diet, physical activity and anthropometry were recorded using standardised techniques. Blood haemoglobin, glucose and total cholesterol were determined. Risk factors were diagnosed using current guidelines. Descriptive statistics are reported. Stepwise multivariate logistic regression was performed to identify determinants of urban-rural differences. RESULTS In urban women mean body mass index (BMI), waist circumference, waist-hip ratio (WHR), systolic BP, haemoglobin, fasting glucose and cholesterol were significantly greater (p<0.01). Age-adjusted prevalence of risk factors (%) in urban vs rural was of obesity BMI ≥ 25 kg/m(2) (45.6 vs 22.5), truncal obesity WHR>0.9 (44.3 vs 13.0), hypertension (37.5 vs 29.3), hypercholesterolemia ≥ 200 mg/dl (27.7 vs 13.5), and diabetes (15.1 vs 4.3) greater whilst any tobacco use (19.6 vs 41.6) or smoking lower. Significant determinants of urban-rural differences were greater income and literacy, dietary fats, low physical activity, obesity and truncal obesity (p<0.01). CONCLUSIONS Greater prevalence of CVD risk factors in urban middle-aged women is explained by greater income and literacy, dietary fat, low physical activity and obesity.

Epicatechin ameliorates ionising radiation-induced oxidative stress in mouse liver

Abstract The current study was intended to evaluate the hepatoprotective effect of Epicatechin (EC) against radiation-induced oxidative stress, in terms of inflammation and lipid peroxidation. Swiss albino mice were administered with EC (15 mg/kg body weight) for three consecutive days before exposing them to a single dose of 5-Gy 60Co gamma (γ) irradiation. Mice were necropsied and livers were taken for immunohistochemistry, western blot analysis and biochemical tests for the detection of markers of hepatic oxidative stress. Nuclear translocation of nuclear factor kappa B (NF-κB) and lipid peroxidation were increased whereas the activities of superoxide dismutase (SOD) and catalase (CAT), reduced glutathione (GSH) content and ferric reducing antioxidant power (FRAP) were diminished upon radiation exposure compared to control. Translocation of NF-κB from cytoplasm to nucleus and lipid peroxidation were found to be inhibited whereas an increase in SOD, CAT, GSH and FRAP was observed in the mice treated with EC prior to irradiation. Thus, pre-treatment with EC offers protection against γ-radiation induced hepatic alterations.

Naringin inhibits gamma radiation-induced oxidative DNA damage and inflammation, by modulating p53 and NF-κB signaling pathways in murine splenocytes

Abstract The adverse effects of ionizing radiation occur due to the generation of reactive oxygen species (ROS). The aim of this study was to identify the protective effects of naringin (NG), a citrus flavonoid, on ionizing radiation (IR)-induced differential stress response, with an exploration of the mechanisms involved in this process. Isolated murine splenocytes were incubated in the presence and in the absence of different concentrations of NG (50 and 100 μM) for 1 h prior to 6 Gy γ-irradiation, and the molecular mechanisms of action were determined through biochemical, immunoblot, flow cytometric, and immunofluorescence studies. Pretreatment with NG significantly prevented IR-induced intracellular ROS generation, thereby preventing the formation of cellular TBARS and the development of cellular nitrite. NG significantly reduced nuclear DNA damage with respect to the irradiated splenocytes, through the inhibition of DNA-PKcs and p-γH2AX. The reduced cell viability as a result of irradiation was recovered by NG through modulation of the redox-regulated cell signaling system. NG pretreatment resulted in significant inhibition of IR-induced G1/S phase cell cycle arrest through the modulation of p53-dependent p21/WAF1, cyclin E, and CDK2 activation. The results also demonstrated that NG blocked the IR-induced p38 function and reversed IR-mediated differential stress response through inhibition of the NF-κB pathway. Thus, the p38/NF-κB pathway participated in the IR-induced inflammatory development, leading to upregulation of CRP, MCP-1, and iNOS2 gene expression. However, NG pretreatment reversed the inflammatory development through downregulation of NF-κB, and regulated the expression of CRP, MCP-1, and iNOS2. The above results provide a theoretical basis for the preventive use of NG against radiation-induced multiple cellular anomalies.

Modulatory role of quercetin against gamma radiation-mediated biochemical and morphological alterations of red blood cells

Abstract Purpose: The present work was intended to evaluate the radioprotective effect of quercetin against gamma radiation-induced oxidative stress on red blood cells (RBC). Materials and methods: Swiss albino male mice were treated with quercetin (100 mg/kg body wt) for three consecutive days prior to 5 Gy 60Co-gamma irradiation. RBC was isolated to estimate the level of intracellular reactive oxygen species (ROS), membrane lipid peroxidation (LPO), intracellular reduced glutathione (GSH), mean corpuscular hemoglobin concentration (MCHC), osmotic fragility and morphological alterations by atomic force microscope (AFM). Results: Irradiation increased intracellular ROS and membrane LPO whereas it decreased the intracellular GSH. Quercetin pretreatment ameliorated these alterations. The MCHC value decreased after irradiation whereas quercetin pretreatment restored it. The average osmotic fragility (H50) and the maximum rate of hemolysis (dH/dC)max increased after irradiation. Quercetin pretreatment decreased the H50 and (dH/dC)max. The AFM study showed that irradiation transformed RBC from biconcave to echinocytes, increased their surface roughness and decreased the vertical distance whereas pretreatment of quercetin significantly prevented both the alterations. Conclusions: Gamma radiation produced ROS and LPO which rendered oxidative stress and ultimately damaged RBC whereas quercetin ameliorated these changes and protected RBC from radiation-mediated damage.

Migrating husbands and changing cardiovascular risk factors in the wife: a cross sectional study in Asian Indian women

Objective The authors studied the influence of migration of husband on cardiovascular risk factors in Asian Indian women. Methods Population-based studies in women aged 35–70 years were performed in four urban and five rural locations. 4608 (rural 2604 and urban 2004) of the targeted 8000 (57%) were enrolled. Demographic details, lifestyle factors, anthropometry, fasting glucose and cholesterol were measured. Multivariate logistic and quadratic regression was performed to compare influence of migration and its duration on prevalence of risk factors. Results Details of migration were available in 4573 women (rural 2267, rural–urban migrants 455, urban 1552 and urban–rural migrants 299). Majority were married, and illiteracy was high. Median (interquartile) duration of residence in urban locations among rural–urban migrants was 9 (4–18) years and in rural areas for urban–rural migrants 23 (18–30) years. In rural, rural–urban migrants, urban and urban–rural migrants, age-adjusted prevalence (%) of risk factors was tobacco use 41.9, 22.7, 18.8 and 38.1; sedentary lifestyle 69.7, 82.0, 79.9 and 74.6; high-fat diet 33.3, 54.2, 66.1 and 61.1; overweight 21.3, 42.7, 46.3 and 29.7; large waist 8.5, 38.5, 29.2 and 29.2; hypertension 30.4, 49.4, 47.7 and 38.4; hypercholesterolaemia 14.4, 31.3, 26.6 and 9.1 and diabetes 3.9, 15.8, 14.9 and 8.4, respectively (p<0.001). In rural–urban migrants, there was a significant correlation of duration of migration with waist size, waist-to-hip ratio and systolic blood pressure (quadratic regression, p<0.001). Association of risk factors with migration remained significant, though attenuated, after adjustment for socioeconomic, lifestyle and obesity variables (logistic regression, p<0.01). Conclusions Compared with rural women, rural–urban migrants and urban have significantly greater cardiometabolic risk factors. Prevalence is lower in urban–rural migrants. There is significant correlation of duration of migration with obesity and blood pressure. Differences are attenuated after adjusting for social and lifestyle variables.

Associação entre marcadores inflamatórios e fatores de risco cardiovascular em mulheres de Kolkata, W.B, Índia

BACKGROUND Recent research has focused on the use of inflammatory biomarkers in the prediction of cardiovascular risk. However, information is scant regarding the association between these inflammatory markers with other cardiovascular risk factors in Asian Indians, particularly in women. OBJECTIVE To explore the association between inflammatory markers such as high-sensitivity C-reactive protein (hs-CRP) and white blood cell (WBC) count and cardiovascular risk factors such as overall and central adiposity, blood pressure, lipid and lipoprotein variables and fasting glucose. METHODS We conducted a cross-sectional analysis on 100 women aged 35-80 years. Participants were selected following cluster sampling methodology from 12 different randomly selected urban wards of Kolkata Municipal Corporation. RESULTS Hs-CRP has a significant association with body mass index (BMI) ( p < 0.001) and waist circumference (WC) (p = 0.002). Significant inverse associations were observed between high-density lipoprotein cholesterol (HDL-C) and both inflammatory markers, hs-CRP (p = 0.031) and WBC count, (p = 0.014). Apolipoprotein A1 (Apo A1) was also negatively associated with hs-CRP. WBC count has significant correlation with fasting glucose and total cholesterol (TC) /HDL-C ratio. Using logistic regression, adjusting for age, BMI (odds ratio/OR, 1.186; confidence interval/CI, 1.046-1.345; p = 0.008) and WC (OR, 1.045; CI, 1.005-1.087; p = 0.027) were the covariates significantly associated with hs-CRP. CONCLUSION In the present study, risk factors like BMI, WC, and HDL-C and apo A1 show significant association with hs-CRP. WBC count was significantly correlated with HDL-C, fasting glucose, TC/HDL-C ratio in women.FUNDAMENTO: Recientes investigaciones se han concentrado en el uso de biomarcadores inflamatorios en la prevision de riesgo cardiovascular. Entre tanto, la informacion es escasa en relacion a la asociacion entre esos marcadores inflamatorios con otros factores de riesgo cardiovasculares en indios asiaticos, particularmente en mujeres. OBJETIVO: Explorar la asociacion entre marcadores inflamatorios tales como proteina C-reactiva de alta sensibilidad (PCR-as) y recuento de leucocitos (LEU) y factores de riesgo cardiovascular tales como adiposidad general y central, presion arterial, variables lipidicas y lipoproteicas y glucemia de ayuno. METODOS: Condujimos un analisis transversal de 100 mujeres con edad entre 35-80 anos. Las participantes fueron seleccionadas a traves de la metodologia de muestreo por cluster, de 12 distritos urbanos seleccionadas al azar en la Corporacion Municipal de Kolkata, India. RESULTADOS: La PCR-as presento una asociacion significativa con el indice de masa corporal (IMC) (r=0,373, p<0,001) y circunferencia de la cintura (CCI) (r=0,301, p=0,002). Asociaciones significativas inversas fueron observadas entre la lipoproteina de alta densidad colesterol (HDL-c) y ambos marcadores inflamatorios (r= -0,220, p=0,031 y r= -0,247, p=0,014 para PCR-as y LEU, respectivamente). La apo-lipoproteina A1 (Apo A1) tambien estaba negativamente asociada con la PCR-as (r= -0,237, p=0,031). El recuento de leucocitos presento una correlacion significativa con la glucemia de ayuno (r=0,253, p=0,011) y la razon colesterol total (CT) /HDL-C (r=0,284, p=0,004). Usando regresion logistica ajustada para edad, IMC (odds ratio/OR, 1,186; intervalo de confianza/IC, 1,046-1,345; p=0,008) y LEU (OR, 1,045; IC, 1,005-1,087; p=0,027) fueron las covariantes significativamente asociadas con la PCR-as. CONCLUSION: En el presente estudio, los factores de riesgo tales como IMC, CCI y HDL-c y Apo-A1 mostraron una asociacion significativa con PCR-as. El recuento de leucocitos estaba significativamente asociado a los niveles de HDL-c, glucemia de ayuno, razon CT/HDL-c en mujeres.