Immunomodulatory Role of Ocimum gratissimum and Ascorbic Acid against Nicotine-Induced Murine Peritoneal Macrophages In Vitro

The aim of this present study was to evaluate the immune functions and immune responses in nicotine-induced (10 mM) macrophages and concurrently establish the immunomodulatory role of aqueous extract of Ocimum gratissimum (Ae-Og) and ascorbic acid. In this study, nitrite generations and some phenotype functions by macrophages were studied. Beside that, release of Th1 cytokines (TNF-α, IL-12) and Th2 cytokines (IL-10, TGF-β) was measured by ELISA, and the expression of these cytokines at mRNA level was analyzed by real-time PCR. Ae-Og, at a dose of 10 μg/mL, significantly reduced the nicotine-induced NO generation and iNOSII expression. Similar kinds of response were observed with supplementation of ascorbic acid (0.01 mM). The administration of Ae-Og and ascorbic acid increased the decreased adherence, chemotaxis, phagocytosis, and intracellular killing of bacteria in nicotine-treated macrophages. Ae-Og and ascorbic acid were found to protect the murine peritoneal macrophages through downregulation of Th1 cytokines in nicotine-treated macrophages with concurrent activation of Th2 responses. These findings strongly enhanced our understanding of the molecular mechanism leading to nicotine-induced suppression of immune functions and provide additional rationale for application of anti-inflammatory therapeutic approaches by O. gratissimum and ascorbic acid for different inflammatory disease prevention and treatment during nicotine toxicity

Amelioratory Effect of Nanoconjugated Vancomycin on Spleen during VRSA-Induced Oxidative Stress

Objective. The aim of the present study was to evaluate the possible antioxidant effects of nanoconjugated vancomycin against VRSA infection on select makers of oxidative damage and antioxidant status in spleen. Methods. A coagulase-positive VRSA strain was used for this study. VRSA infection was developed in Swiss mice by intraperitoneal injection of 5 × 106 CFU/mL bacterial solutions. VRSA-infected mice were treated with nanoconjugated vancomycin at its effective dose for 10 days. After decapitation, blood was used for determination of viable bacteria count and spleen was excised from control and experimental groups, homogenized and used for different biochemical estimations. Results. Nitrate level, myeloperoxidase activity, lipid peroxidation, protein oxidation, oxidized glutathione, and DNA fragmentation level were increased significantly (P < 0.05) in spleen of VRSA-infected group as compared to control group, and reduced glutathione level, activity of SOD, CAT, GPx, GR, and GST were decreased significantly (P < 0.05); which were increased or decreased significantly (P < 0.05) near to normal in nanoconjugated vancomycin-treated group. Conclusion. These findings suggest the potential use and beneficial role of nanoconjugated vancomycin against VRSA-infection-induced oxidative stress and DNA damage in spleen

Age associated oxidative damage in lymphocytes

Lymphocytes are an important immunological cell and have been played a significant role in acquired immune system; hence, may play in pivotal role in immunosenescence. Oxidative stress has been reported to increase in elderly subjects, possibly arising from an uncontrolled production of free radicals with aging and decreased antioxidant defenses. This study was aimed to evaluate the level of lipid-protein damage and antioxidant status in lymphocytes of healthy individuals to correlate between oxidative damage with the aging process. Twenty healthy individuals of each age group (11–20; 21–30; 31–40; 41–50; and 51–60 years) were selected randomly. Blood samples were drawn by medical practitioner and lymphocytes were isolated from blood samples. Malondialdehyde (MDA), protein carbonyls (PC) level were evaluated to determine the lipid and protein damage in lymphocytes. Superoxide dismutase (SOD), catalase (CAT), glutathione and glutathione dependent enzymes were estimated to evaluate the antioxidant status in the lymphocytes. Increased MDA and PC levels strongly support the increased oxidative damage in elderly subject than young subjects. The results indicated that, balance of oxidant and antioxidant systems in lymphocytes shifts in favor of accelerated oxidative damage during aging. Thus oxidative stress in lymphocytes may particular interest in aging and may play important role in immunosenescence

Internalization of Staphylococcus aureus in Lymphocytes Induces Oxidative Stress and DNA Fragmentation: Possible Ameliorative Role of Nanoconjugated Vancomycin

Staphylococcus aureus is the most frequently isolated pathogen causing bloodstream infections, skin and soft tissue infections and pneumonia. Lymphocyte is an important immune cell. The aim of the present paper was to test the ameliorative role of nanoconjugated vancomycin against Vancomycin-sensitive Staphylococcus aureus (VSSA) and vancomycin-resistant Staphylococcus aureus (VRSA) infection-induced oxidative stress in lymphocytes. VSSA and VRSA infections were developed in Swiss mice by intraperitoneal injection of 5 × 106 CFU/mL bacterial solutions. Nanoconjugated vancomycin was adminstrated to VSSA- and VRSA-infected mice at its effective dose for 10 days. Vancomycin was adminstrated to VSSA- and VRSA-infected mice at a similar dose, respectively, for 10 days. Vancomycin and nanoconjugated vancomycin were adminstrated to normal mice at their effective doses for 10 days. The result of this study reveals that in vivo VSSA and VRSA infection significantly increases the level of lipid peroxidation, protein oxidation, oxidized glutathione level, nitrite generation, nitrite release, and DNA damage and decreases the level of reduced glutathione, antioxidant enzyme status, and glutathione-dependent enzymes as compared to control group, which were increased or decreased significantly near to normal in nanoconjugated vancomycin-treated group. These findings suggest the potential use and beneficial role of nanoconjugated vancomycin against VSSA and VRSA infection-induced oxidative stress in lymphocytes

In vitro Staphylococcus aureus–induced oxidative stress in mice murine peritoneal macrophages: a duration–dependent approach

Objective: To evaluate the free radical generation and status of the antioxidant enzymes in murine peritoneal macrophage during in vitro vancomycin sensitive Staphylococcus aureus (VSSA) treatment with different time interval. Methods: Peritoneal macrophages were treated with 5×106 CFU/mL VSSA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity, nitric oxide generation, antioxidant enzyme status and components of glutathione cycle were analyzed. Results: Superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity and nitric oxide generation got peak at 3 h, indicating maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during VSSA infection. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-s-transferase activity were decreased significantly (P<0.05) with increasing time of VSSA infection. But the oxidized glutathione level was time dependently increased significantly (P<0.05) in murine peritoneal macrophages. All the changes in peritoneal macrophages after 3 h in vitro VSSA treatment had no significant difference. Conclusions: From this study, it may be summarized that in vitro VSSA infection not only generates excess free radical but also affects the antioxidant status and glutathione cycle in murine peritoneal macrophages

Aqueous extract of Ocimum gratissimum Linn and ascorbic acid ameliorate nicotine–induced cellular damage in murine peritoneal macrophage

AbstractObjectiveTo test the in vitro protective role of aqueous extract of Ocimum gratissimum Linn. (O. gratissimum) and ascorbic acid against nicotine-induced murine peritoneal macrophage.MethodsPeritoneal macrophages from mice were treated with nicotine (10 mM), nicotine (10 mM) with aqueous extract of O. gratissimum (1 to 25 μg/mL), and nicotine (10 mM) with ascorbic acid (0.01 mM) for 12 h in cell culture media, while the control group was treated with culture media. Levels of free radical generation, lipid peroxidation, protein carbonyls, oxidized glutathione levels and DNA damage were observed and compared.ResultsPhytochemical analysis of aqueous extract has shown high amount of phenolics and flavonoids compound present in it. The significantly increased free radical generation, lipid peroxidation, protein carbonyls, oxidized glutathione levels and DNA damage were observed in nicotine-treated group as compared to the control group; those were significantly reduced in aqueous extract of O. gratissimum and ascorbic acid supplemented groups. Moreover, significantly reduced antioxidant status in nicotine exposed murine peritoneal macrophage was effectively ameliorated by these two products. Among the different concentration of aqueous extract of O. gratissimum, the maximum protective effect was observed at 10 μg/mL which does not produce any significant change in the normal cell.ConclusionsThese findings suggest the potential use and beneficial role of O. gratissimum as a modulator of nicotine-induced cellular damage in murine peritoneal macrophage

In Vitro Time Dependent Nicotine-Induced Free Radical Generation and Status of Glutathione Cycle in Murine Peritoneal Macrophage

: Nicotine is an alkaloid and the precursor to many tobacco carcinogens present in various tobacco products. The aim of the present study was to evaluate the free radical generation and status of the glutathione cycle in murine peritoneal macrophage during in vitro nicotine exposure with different time interval. Peritoneal macrophages were treated with 10mM nicotine in vitro for different time interval (3, 6, 9, 12, 15, 18, & 24h) and super oxide anion generation and components of glutathione cycle were analyzed. Super oxide anion generation, and NADPH oxidase activity got peak at 12hr, indicates maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during nicotine treatment. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase activity were decreased significantly (p<0.05) with increasing time of nicotine treatment. But the oxidized glutathione level was time dependently increased significantly (p<0.05) in murine peritoneal macrophages. The redox ratio also decreased significantly (p<0.05) at all time of nicotine treatment. All the changes in peritoneal macrophages after 12hr in vitro nicotine treatment had no significant difference. From this study, it may be summarized that nicotine not only generates excess free radical but also affect the glutathione cycle in murine peritoneal macrophage