background: In an attempt to investigate the role of oxidants in the activation of phospholipase $A_2$(PLA$_2$) and endogenous oxidative stress in the lung. acute inflammatory lung injury was induced by the instillation of hydrogen peroxide into the trachea of Sprague-Dawley rats. Material and Method: To prove the hypothesis thats released oxidants from neutrophils activate the PLA$_2$ retrogradely, activities of PLA$_2$ and lysoplatelet activating factor acetyltransferase(lysoPAF AT) were assayed i hours after instillation of hydrogen peroxide. In addition, to confirm the impairing effects of the activation of PLA$_2$ associated with endogenous oxidative stress, lung weight/body weight ratio(L$\times$10$^{-3}$ B), protein contents(mg/two lungs) in bronchoalveolar lavage(BAL) were measured. As neutrophilic respiratory burst has been known to play a pivotal role in the genesis of endogenous oxidative stress associated with acute inflammatory lung injury, BAL neutrophils counts and level of lung myelperoxidase(MPO) were measured after hydorgen peroxide insult. Morphological and histochemical studies were also performed to identify the effect of the endogenous oxidative stress. Result: Five hours after hydrogen peroxide instillation, lungs showed marked infiltration of neutrophils and increased weight. Protein contents in BAL increased significantly compared to those of normal rats. PLA$_2$ activity was enhanced in the hydrogen peroxide instilled group. Interestingly, the accelerated production of platelet activating factor(PAF) was confirmed by the increased activity of lysoPAF AT in the $H_2O$$_2$ employed lung. Morphologically, light microscopic findings of lungs after instillation of hydrogen peroxide showed atelectasis and infiltration of inflammatory cells, which was thought to be caused by lipid mediators produced by PLA$_2$ activation. In cerium chloride cytochemical electron microscopy, dense deposits of cerrous perhydroxide were identified. In contrast, no deposit of cerrous perhydroxide was found in the normal lung.
Purpose: Current studies have demonstrated the neuroprotective effects of 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) in many animal models of brain injury, including hypoxic-ischemic (HI) encephlopathy, trauma and excitotoxicity, but limited data are available for those during the neonatal periods. Here we investigated whether CNQX can protect the developing rat brain from HI injury via mediation of nitric oxide synthase. Methods: In an in vivo model, left carotid artery ligation was done in 7-day-old Sprague-Dawley (SD) rat pups. The animals were divided into six groups; normoxia (N), hypoxia (H), hypoxia with sham-operation (HS), hypoxia with operation (HO), HO treated with vehicle (HV), and HO treated with CNQX at a dose of 10 mg/kg (HC). Hypoxia was made by exposure to a 2 hr period in the hypoxic chamber (92% $N_2$, 8% $O_2$). In an in vitro model, embryonic cortical neuronal cell culture of SD rats at 18-day gestation was done. The cultured cells were divided into three groups: normoxia (N), hypoxia (H), and hypoxia treated with CNQX (HC). The N group was prepared in 5% $CO_2$ incubators and the other groups were placed in 1% $O_2$) incubators (94% $N_2$, 5% $CO_2$) for 16 hr. Results: In the in vitvo and in vivo models, the expressions of iNOS and eNOS were reduced in the hypoxia group when compared to the normoxia group, whereas they were increased in the CNQX-treated group compared to the hypoxia group. In contrast, the expression of nNOS was showed reversely. Conclusion: CNQX has neuroprotective property over perinatal HI brain injury via mediation of nitric oxide synthase.
Journal of the Korean Society of Food Science and Nutrition
/
v.24
no.6
/
pp.867-873
/
1995
This study was done ot investigate the effect of chronic alcohol feeding and acetylaminofluorene(2-AAF) treatment on hepatic mitochondrial ATPase activity andmembrane lipid composition. Male Sprague-Dawley rats, weighing 120~125g, were fed for 6 weeks on a liquid diet containing 35% of calories as ethanol. After 4 weeks of experiment diet feeding, 2-AAF(100mg/kg body weight) was injected twice a week intraperitoneally. Body weight and percent liver weight per body weight were significantly changed by ethanol feeding. Hepatic mitochondrial ATPase activity significantly decreased by ethanol feedings but not by 2-AAF treatment. In comparison to control, the ATPase activity of ethanol-AAF group decreased 29.3%. Since phospholipid(PL) content of mitochondria has an interaction effect between ethanol and 2-AAF treatment, 2-AAF treatment significantly increased phospholipid content in only ethanol fed group. Total cholesterol(C) level of mitochondria significantly increased by ethanol feeding. Consequently C/PL ratio of ethanol group was significantly higher than that of control group. The analysis of mitochondrial PL composition showed that cardiolipin(CL) significantly increased by 2-AFF treatment in control group. Phosphatidyl choline(PC) significantly increased by ethanol feeding, whereas PC significanlty decreased and phosphatidyl ethanolamine(PE) significantly increased by 2-AAF treatment. 2-AAF treatment also showed a significant increase in PE/PC ratio. Fatty acid patterns of mitochondria were also changed by either ethanol or 2-AAF although the severity of the changes was not great. These data suggest that the reduced mitochondrial ATPase activity in ethanol-AAF group may be a consequence of a changes in mitochondrial membrane lipid composition such as PE/PC ratio, C/PL ration and fatty acid patterns.
Park, Sung-Hye;Shin, Eon-Hwan;Park, Sung-Jin;Ran, Jong-Hyun
Journal of the Korean Society of Food Science and Nutrition
/
v.34
no.4
/
pp.476-483
/
2005
This experiment was planned to develop a functional supplement by food resources to prevent and lessen the deleterious effects caused by environmental pollutants such as polluted food, air, water and heavy metals. The goal of this study was to investigate the effects of peking-duck extracts supplemented with six kinds of medicinal herbs (DJ) on the intoxication of lead and mercury in rats. Sprague-Dawley rat weighing $150g\pm15g$ g, were randomly assigned to 5 groups, basal diet only (NCG), heavy metal without DJ injection (HCG), heavy metals and DJ (3 mg/mL) injection (HMLD), heavy metal and DJ (30 mg/mL) injection (HMMD), heavy metal and D] (300 mg/mL) injection (HMHD). Mecury (Hg) and lead (Pb) injected at the level of 50 ppm for 17 days. Also DJ oral feeding was continued for 31 days. The result of this study were as follows; Food intake and body weight gain in heavy metal administered groups were lower than those of control group (NCG). The activities of GOT, GPT and BUN level were significantly reduced in DJ-treated groups as compared to HCG. DJ was shown to suppress the accumulation of Hg and Pb in serum. The results suggest that DJ might have protective effect on Hg and Pb intoxication.
The Journal of the Korean Society for Microbiology
/
v.21
no.1
/
pp.133-144
/
1986
This study was undertaken to assess the effect of ginseng administration on T lymphocyte induced local xenogenic graft-versus-host(GVM) reactions which were induced with thymocyte, spleen cell and lymph node cell of ICR mice. Mice received daily 10mg of 70% alcohol ginseng extract oral1y for 100days and control mice remained untreated for the same period of time. The cells from donor mice were injected intradermally into the closely shaven abdominal skin of Sprague-Dawley rats for GVH tests. The thymocyte from control(ginseng-untreated) mice showed a negative local GVH reaction, whereas thymocyte from experimental(ginseng-treated) mice showed a positive reaction with the rate of 17.4%. When spleen cells were injected, the incidence of positive local GVH reaction was 66.7% among ginseng-treated mice, as opposed to incidence of 45.5% of positive local GVH reaction among control mice. The incidence of positive local GVH reaction of the lymph node cells when injected into a recipient was 71.4% among ginseng-treated mice as compared with that of 18.9% among control mice. The relationship between spleen cell inoculum and intensity of the local GVH reaction was assessed in ginseng-untreated mice. The intensity of GVH reaction clearly appears to be dose related. In ginseng-treated mice, a minimum of $1{\times}10^7$ spleen cell was required for production of positive local GVH reaction with almost linear relationship up to an inoculum of $5{\times}10^8$ cells. In control mice, however, a minimum of $1{\times}10^8$ spleen cells was required for positive GVH reaction. These results strongly suggest that the ginseng administration augments significantly the local xenogenic GVH reaction which was used to assess T lymphocyte function and immunocompetence of mice and in addition to this, these results appear to support previous suggestions that the local GVH reaction consitutes a qualitative test of the functional activity of T lymphocytes. These results may be the first to induce local GVH reaction, employing rats as recipient and mice as donor. This study was also desingned to investigate some of the effects of ginseng extract on lymphocyte-macrophage interactions. This was accomplished by in vitro quantification of 1) migratory inhibitory factor(MIF) synthetic capacity of splenic lymphocytes in mice previously primed with ginseng 2) MIF responsiveness of mouse peritoneal macrophages or chicken peripheral leucocytes under the presence of ginseng extract 3) migration ability of chicken peripheral leucocytes by direct stimulation of ginseng extract or ginseng saponin and 4) immunosuppressive effects of immunosuppressants such as cyclophosphamide, cyclosporin A or dexamethasone. Mice divided equally into the ginseng and the saline groups, which received intraperitoneally daily 0.2ml of ginseng absolute alcohol-extract(5mg/ml) and same amount of saline for 15 days, respectively. The cellular immune responsiveness of these mice was assayed 15 days after ginseng pretreatment. Splenic lymphocytes of mice treated with ginseng, when stimulated with sensitized specific-antigen such as sheep red blood cells or toxoplasmin, or with polyclonal activator concanavalin A, produced significantly more MIF than those of control saline group. MIF responsiveness of normal mouse macrophages was significantly augmented when assayed under the presence of ginseng extract (1mg/ml). The migratory ability of normal chicken leucocytes in the absence of MIF was significantly decreased by the stimulation of ginseng extract alone. MIF response was significantly decreased by immunosuppressants and this impaired response was not restored by ginseng pretreatment. This study was additionally performed to evaluate the effect of ginseng on the expulsion of adult Trichinella spiralis in mice. ICR mice were infected experimentally by esophageal incubation of 300 T. spiralis infective muscle larvae prepared by acid-pepsin digestion of infected mice. and received oral administration of 70% alcohol ginseng extract(10mg/mouse/day) for the indicated days plus 4 days before infection. At various times after infection, the number of adult T. spiralis worms in small intestines was determined. Interestingly, ginseng-treatment was accompanied by accelerated expulson of T. spiralis. These results led to the conclusion that Panax ginseng caused some enhancing effect on GVH reaction, macrophage migration inhibition reaction and expulsion of T. spiralis. In addition these results suggested that the mechanisms responsible for this enhancement of ginseng may be chiefly or partially due to nonspecific stimulation of cell-mediated immune response.
Purpose: Erythropoietin (EPO) has neuroprotective effects in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity. Current studies have demonstrated the neuroprotective effects of EPO, but limited data are available for the neonatal periods. Here in we investigated whether recombinant human EPO (rHuEPO) can protect the developing rat brain from HI injury via modulation of NMDA receptors. Methods: In an in vitro model, embryonic cortical neuronal cell cultures from Sprague-Dawley (SD) rats at 19-days gestation were established. The cultured cells were divided into five groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated (H+E1, H+ E10, and H+E100) groups. To estimate cell viability and growth, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was done. In an in vivo model, left carotid artery ligation was performed on 7-day-old SD rat pups. The animals were divided into six groups; normoxia control (NC), normoxia Sham-operated (NS), hypoxia-ischemia only (H), hypoxia-ischemia+vehicle (HV), hypoxia-ischemia+rHuEPO before a HI injury (HE-B), and hypoxia-ischemia+rHuEPO after a HI injury (HE-A). The morphologic changes following brain injuries were noted using hematoxylin and eosin (H/E) staining. Real-time PCR using primers of subunits of NMDA receptors (NR1, NR2A, NR2B, NR2C and NR2D) mRNA were performed. Results: Cell viability in the H group was decreased to less than 60% of that in the N group. In the H+E1 and H+E10 groups, cell viability was increased to >80% of the N group, but cell viability in the H+E100 group did not recover. The percentage of the left hemisphere area compared the to the right hemisphere area were 98.9% in the NC group, 99.1% in the NS group, 57.1% in the H group, 57.0% in the HV group, 87.6% in the HE-B group, and 91.6% in the HE-A group. Real-time PCR analysis of the expressions of subunits of NMDA receptors mRNAs in the in vitro and in vivo neonatal HI brain injuries generally revealed that the expression in the H group was decreased compared to the N group and the expressions in the rHuEPO-treated groups was increased compared to the H group. Conclusion: rHuEPO has neuroprotective property in perinatal HI brain injury via modulation of N-methyl-D-aspartate receptors.
To detect ischemic tissue in experimental model of cerebral ischemia made by middle cerebral artery(MCA)-occlusion, we acquired triple image of $^{99m}Tc$-glucarate, [$^{18}F$]fluoro-deoxyglucose (FDG), and 2,3,5- triphenyltetrazolium (TTC) staining. We made cerebral infarction either with reperfusion (after occlusion of 2 hours) or without reperfusion in 10 Sprague-Dawley rats by inserting thread to MCA through internal carotid artery. After 22 hours, we injected 740 MBq of $^{99m}Tc$-glucarate and 55.5 MBq of [$^{18}F$]FDG through tail vein. Each 1 mm slice of rat brains was frozen and exposed to imaging plate for 20 minutes in freezer to get an [$^{18}F$]FDG image. After 20 hours enough to fade radioactivity of [$^{18}F$]FDG, the slices were again imaged by BAS1500 for $^{99m}Tc$-glucarate uptake. Finally, these brain tissues were stained with TTC. Semi-quantitative visual analysis was done by grading 0 to 3 points according to the degree of uptakes($^{99m}Tc$-glucarate) and decreased uptakes([$^{18}F$]FDG and TTC). Ten rats survived with neurologic symptoms. TTC staining confirmed the development of infarction. The size of the infarction was relatively larger in the group without reperfusion. [$^{18}F$]FDG images were similar to TTC-stained images. However, we found regions with intermediate uptake which were not stained with TTC. We found regions with intermediate [$^{18}F$]FDG uptake where TTC staining was normal. $^{99m}Tc$-glucarate uptake was round only in TTC non-stained region. In the TTC stained regions, there were no uptake of $^{99m}Tc$-glucarate. We could not find clear relation between $^{99m}Tc$-glucarate uptake with [$^{18}F$]FDG uptake. This was partly because percent uptake of $^{99m}Tc$-glucarate was so small (less than 1 percent of injected dose) and because there were quite heterogeneity of patterns of [$^{18}F$]FDG uptake and TTC. With these findings, we could conclude that $^{99m}Tc$-glucarate were taken up only in part of ischemic tissues which were proven to be nonviable. The establishment of MCA-occluded rat model with or without reperfusion and triple imaging for $^{99m}Tc,\;^{18}F$ and TTC helped the characterization of $^{99m}Tc$-glucarate uptakes. Further work is needed to clarify the meaning or diversities or [$^{18}F$]FDG and TTC and their relation with $^{99m}Tc$-glucarate.
This study was conducted to determine the antiseptic effect of stabilized chlorine dioxide (S-$ClO_2$) on muscle tissue of rats. Skeletal muscle of 8-week old Sprague-Dawley rats was used. Light and transmission electron microscopic findings were observed in the control group, which was not treated with stabilized chlorine dioxide, and in the experimental group, which was treated with a stabilized chlorine dioxide powder in aqueous solution. According to the LM and TEM observations, the day 1 control group showed the initiation of endomysium collapse resulting in an unclear boundary of muscle fibers, and partial collapse of the mitochondrial membranes. All endomysium had collapsed, and bacteria were observed among muscle fibers in the day 2 and later groups. Shapes of muscles were not distinguishable in day 3 or later groups. In contrast, the day 1 and 3 experimental groups revealed detailed structure of typical muscles, but partial collapse of the mitochondrial membranes was observed in the day 3 and later groups. Subsequently, connective tissues collapsed and structures in the shape of concentric circles were observed. In summary, the day 1 control group showed the initial collapse of tissues, and shapes were not distinguishable in the day 3 and later groups because most of the tissues had collapsed. In contrast, the day 3 experimental group showed partial collapse, but the overall shapes of muscles were maintained as time went on, confirming the antiseptic effect of stabilized chlorine dioxide on muscles.
Kang, Min-Jung;Hwang, Cho-Rong;Lee, Soo Jung;Shin, Jung-Hye
Journal of Life Science
/
v.24
no.7
/
pp.728-736
/
2014
The purpose of this study was to improve and strengthen the function of eel extract prepared with 5 brix eel extract (EE), 5 brix eel and plant mix (black garlic, ginseng, black jujube) 0.35 ml extracts prepared and treated with the extract (EIM-1), and 0.7 ml (EIM-2) divided group. The extracts were administered to rats for five weeks during running training, and the lipid profiles and antioxidant enzyme activities were tested. The lipid content in liver and serum were lower than the normal group difference was not significant between groups. Serum total cholesterol was lower in the experimental group than the control group the mixed extract significantly lower level. HDL-cholesterol levels in the eel extract and eel mixed extract significantly increased by feeding the EIM-1 is 2.0 times, EIM-2 is increased by 2.3 times. Liver glycogen content in the experimental group performed the exercise group compared with the normal control group was significantly lower than in EIM is significantly higher than the control group. The TBARS content in the liver and serum was significantly higher than the normal group was lower than the control group. GOT and GPT were significantly decreased compared to the control group. Hepatic catalase activity was significantly increased in the EIM-1 group, and SOD and GSH-px activities were increased in the EIM-1 and EIM-2 groups. Supplementation with the eel and plant mix extract increased the activities of antioxidant enzymes. Thus, intake of the eel and plant mix extract could improve the antioxidant status and combat different types of oxidative stress.
This study investigated the hepatoprotective effects of an ethanol extract of lotus root (LRE) on alcohol-induced liver damage in rat. Sprague-Dawley rae weighing $100{\sim}150g$, were divided into 6 groups: basal diet group (BD), alcohol (35% 10 mL/kg/day) teated stoup (ET), LRE 200 mg/kg/day teated group (BD-LREL). LRE 400 mg/kg/day treated group (BD-LREH), LRE 200 mg/kg/day and alcohol treated group (ET-LREL), and LRE 400 3mg/kg/day and alcohol teated group (ET-LREH). After the administration, rats were sacrificed to get serum and liver to analyze antioxidant enzyme activity, glutathione and lipid peroxide contents. The body weight gain and feed efficiency ratio were decreased by alcohol administration, however, were gradually increased to a little lower level than the basal diet group by the combined administration of alcohol and LRE. The serum alanine aminotransferase (ALT), asparate aminotransferase (AST) and alkaline phosphatase (ALP) activities that were elevated by alcohol were significantly decreased by LRE administration. It was also observed that thiobarbituric acid reactive substances (TBARS) content, xanthine oxidase (XO), superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) activities in liver that were increased by alcohol, were markedly decreased in the combined alcohol and LRE administered groups as compared with the alcohol administrated group. These effect of LRE within the alcohol groups were in a dose-dependent manner. The glutathione (GSH) content in liver was decreased by alcohol administration, however, increased after administering LRE. Teken together, these result suggest that ethanol extract of lotus root may have a possible protective effect on liver function in hepatotoxicity-induced rat by alcohol administration.
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