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Biomedical and Environmental Sciences ; (12): 54-70, 2024.
Article in English | WPRIM | ID: wpr-1007908


OBJECTIVE@#The aim of this study is to explore the potential modulatory role of quercetin against Endotoxin or lipopolysaccharide (LPS) induced septic cardiac dysfunction.@*METHODS@#Specific pathogen-free chicken embryos ( n = 120) were allocated untreated control, phosphate buffer solution (PBS) vehicle, PBS with ethanol vehicle, LPS (500 ng/egg), LPS with quercetin treatment (10, 20, or 40 nmol/egg, respectively), Quercetin groups (10, 20, or 40 nmol/egg). Fifteen-day-old embryonated eggs were inoculated with abovementioned solutions via the allantoic cavity. At embryonic day 19, the hearts of the embryos were collected for histopathological examination, RNA extraction, real-time polymerase chain reaction, immunohistochemical investigations, and Western blotting.@*RESULTS@#They demonstrated that the heart presented inflammatory responses after LPS induction. The LPS-induced higher mRNA expressions of inflammation-related factors (TLR4, TNFα, MYD88, NF-κB1, IFNγ, IL-1β, IL-8, IL-6, IL-10, p38, MMP3, and MMP9) were blocked by quercetin with three dosages. Quercetin significantly decreased immunopositivity to TLR4 and MMP9 in the treatment group when compared with the LPS group. Quercetin significantly decreased protein expressions of TLR4, IFNγ, MMP3, and MMP9 when compared with the LPS group. Quercetin treatment prevented LPS-induced increase in the mRNA expression of Claudin 1 and ZO-1, and significantly decreased protein expression of claudin 1 when compared with the LPS group. Quercetin significantly downregulated autophagy-related gene expressions (PPARα, SGLT1, APOA4, AMPKα1, AMPKα2, ATG5, ATG7, Beclin-1, and LC3B) and programmed cell death (Fas, Bcl-2, CASP1, CASP12, CASP3, and RIPK1) after LPS induction. Quercetin significantly decreased immunopositivity to APOA4, AMPKα2, and LC3-II/LC3-I in the treatment group when compared with the LPS group. Quercetin significantly decreased protein expressions of AMPKα1, LC3-I, and LC3-II. Quercetin significantly decreased the protein expression to CASP1 and CASP3 by immunohistochemical investigation or Western blotting in treatment group when compared with LPS group.@*CONCLUSION@#Quercetin alleviates cardiac inflammation induced by LPS through modulating autophagy, programmed cell death, and myocardiocytes permeability.

Chick Embryo , Animals , Quercetin/therapeutic use , Lipopolysaccharides/toxicity , Matrix Metalloproteinase 9 , Caspase 3 , Matrix Metalloproteinase 3 , Toll-Like Receptor 4 , Claudin-1 , Inflammation/metabolism , Apoptosis , RNA, Messenger , Autophagy , NF-kappa B
Journal of Zhejiang University. Science. B ; (12): 430-441, 2023.
Article in English | WPRIM | ID: wpr-982383


Early weaned piglets suffer from oxidative stress and enteral infection, which usually results in gut microbial dysbiosis, serve diarrhea, and even death. Rice bran oil (RBO), a polyphenol-enriched by-product of rice processing, has been shown to have antioxidant and anti-inflammatory properties both in vivo and in vitro. Here, we ascertained the proper RBO supplementation level, and subsequently determined its effects on lipopolysaccharide (LPS)-induced intestinal dysfunction in weaned piglets. A total of 168 piglets were randomly allocated into four groups of seven replicates (42 piglets each group, (21±1) d of age, body weight (7.60±0.04) kg, and half males and half females) and were given basal diet (Ctrl) or basal diet supplemented with 0.01% (mass fraction) RBO (RBO1), 0.02% RBO (RBO2), or 0.03% RBO (RBO3) for 21 d. Then, seven piglets from the Ctrl and the RBO were treated with LPS (100 μg/kg body weight (BW)) as LPS group and RBO+LPS group, respectively. Meanwhile, seven piglets from the Ctrl were treated with the saline vehicle (Ctrl group). Four hours later, all treated piglets were sacrificed for taking samples of plasma, jejunum tissues, and feces. The results showed that 0.02% was the optimal dose of dietary RBO supplementation based on diarrhea, average daily gain, and average daily feed intake indices in early weaning piglets. Furthermore, RBO protected piglets against LPS-induced jejunal epithelium damage, which was indicated by the increases in villus height, villus height/crypt depth ratio, and Claudin-1 levels, as well as a decreased level of jejunal epithelium apoptosis. RBO also improved the antioxidant ability of LPS-challenged piglets, which was indicated by the elevated concentrations of catalase and superoxide dismutase, and increased total antioxidant capacity, as well as the decreased concentrations of diamine oxidase and malondialdehyde in plasma. Meanwhile, RBO improved the immune function of LPS-challenged weaned piglets, which was indicated by elevated immunoglobulin A (IgA), IgM, β‍‍-defensin-1, and lysozyme levels in the plasma. In addition, RBO supplementation improved the LPS challenge-induced dysbiosis of gut microbiota. Particularly, the indices of antioxidant capacity, intestinal damage, and immunity were significantly associated with the RBO-regulated gut microbiota. These findings suggested that 0.02% RBO is a suitable dose to protect against LPS-induced intestinal damage, oxidative stress, and jejunal microbiota dysbiosis in early weaned piglets.

Male , Female , Animals , Swine , Lipopolysaccharides/toxicity , Antioxidants/pharmacology , Rice Bran Oil , Dysbiosis , Dietary Supplements , Diarrhea/veterinary , Weaning , Body Weight
China Journal of Chinese Materia Medica ; (24): 4201-4207, 2023.
Article in Chinese | WPRIM | ID: wpr-1008616


This study aims to explore the neuroprotective effect of bilobalide(BB) and the mechanisms such as inhibiting inflammatory response in macrophage/microglia, promoting neurotrophic factor secretion, and interfering with the activation and differentiation of peripheral CD4~+ T cells. BB of different concentration(12.5, 25, 50, 100 μg·mL~(-1)) was used to treat the RAW264.7 and BV2 cells for 24 h. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and cell counting kit-8(CCK-8) were employed to detect the cytotoxicity of BB and appropriate concentration was selected for further experiment. Lipopolysaccharide(LPS) was applied to elicit inflammation in RAW264.7 and BV2 cells, mouse bone marrow-derived macrophages(BMDMs), and primary microglia, respectively. The effect of BB on cell proliferation and secretion of inflammatory cytokines and neurotrophic factors was detected by enzyme-linked immunosorbent assay(ELISA). Spleen monocytes of C57BL/6 female mice(7-8 weeks old) were isolated, and CD4~+ T cells were separated by magnetic beads under sterile conditions. Th17 cells were induced by CD3/CD28 and the conditioned medium for eliciting the inflammation in BMDMs. The content of IL-17 cytokines in the supernatant was detected by ELISA to determine the effect on the activation and differentiation of CD4~+ T cells. In addition, PC12 cells were incubated with the conditioned medium for eliciting inflammation in BMDMs and primary microglia and the count and morphology of cells were observed. The cytoto-xicity was determined by lactate dehydrogenase(LDH) assay. The result showed that BB with the concentration of 12.5-100 μg·mL~(-1) had no toxicity to RAW264.7 and BV2 cells, and had no significant effect on the activity of cell model with low inflammation. The 50 μg·mL~(-1) BB was selected for further experiment, and the results indicated that BB inhibited LPS-induced secretion of inflammatory cytokines. The experiment on CD4~+ T cells showed that the conditioned medium for LPS-induced inflammation in BMDMs promoted the activation and differentiation of CD4~+ T cells, while the conditioned medium of the experimental group with BB intervention reduced the activation and differentiation of CD4~+ T cells. In addition, BB also enhanced the release of neurotrophic factors from BMDMs and primary microglia. The conditioned medium after BB intervention can significantly reduce the death of PC12 neurons, inhibit neuronal damage, and protect neurons. To sum up, BB plays a neuroprotective role by inhibiting macrophage and microglia-mediated inflammatory response and promoting neurotrophic factors.

Female , Rats , Mice , Animals , Bilobalides/pharmacology , Neuroprotection , Lipopolysaccharides/toxicity , Culture Media, Conditioned/pharmacology , Mice, Inbred C57BL , Macrophages/metabolism , Microglia , Cytokines/metabolism , Nerve Growth Factors/pharmacology , Inflammation/metabolism
Journal of Integrative Medicine ; (12): 274-280, 2022.
Article in English | WPRIM | ID: wpr-929222


OBJECTIVE@#Acute lung injury (ALI) is a serious respiratory dysfunction caused by pathogen or physical invasion. The strong induced inflammation often causes death. Tanshinone IIA (Tan-IIA) is the major constituent of Salvia miltiorrhiza Bunge and has been shown to display anti-inflammatory effects. The aim of the current study was to investigate the effects of Tan-IIA on ALI.@*METHODS@#A murine model of lipopolysaccharide (LPS)-induced ALI was used. The lungs and serum samples of mice were extracted at 3 days after treatment. ALI-induced inflammatory damages were confirmed from cytokine detections and histomorphology observations. Effects of Tan-IIA were investigated using in vivo and in vitro ALI models. Tan-IIA mechanisms were investigated by performing Western blot and flow cytometry experiments. A wound-healing assay was performed to confirm the Tan-IIA function.@*RESULTS@#The cytokine storm induced by LPS treatment was detected at 3 days after LPS treatment, and alveolar epithelial damage and lymphocyte aggregation were observed. Tan-IIA treatment attenuated the LPS-induced inflammation and reduced the levels of inflammatory cytokines released not only by inhibiting neutrophils, but also by macrophage. Moreover, we found that macrophage activation and polarization after LPS treatment were abrogated after applying the Tan-IIA treatment. An in vitro assay also confirmed that including the Tan-IIA supplement increased the relative amount of the M2 subtype and decreased that of M1. Rebalanced macrophages and Tan-IIA inhibited activations of the nuclear factor-κB and hypoxia-inducible factor pathways. Including Tan-IIA and macrophages also improved alveolar epithelial repair by regulating macrophage polarization.@*CONCLUSION@#This study found that while an LPS-induced cytokine storm exacerbated ALI, including Tan-IIA could prevent ALI-induced inflammation and improve the alveolar epithelial repair, and do so by regulating macrophage polarization.

Animals , Mice , Abietanes , Acute Lung Injury/drug therapy , Cytokine Release Syndrome , Cytokines , Inflammation/drug therapy , Lipopolysaccharides/toxicity , Macrophage Activation , Macrophages , Triacetoneamine-N-Oxyl/pharmacology
Chinese Medical Sciences Journal ; (4): 1-14, 2022.
Article in English | WPRIM | ID: wpr-928241


Objective To examine the neuroanatomical substrates underlying the effects of minocycline in alleviating lipopolysaccharide (LPS)-induced neuroinflammation. Methods Forty C57BL/6 male mice were randomly and equally divided into eight groups. Over three conse-cutive days, saline was administered to four groups of mice and minocycline to the other four groups. Immediately after the administration of saline or minocycline on the third day, two groups of mice were additionally injected with saline and the other two groups were injected with LPS. Six or 24 hours after the last injection, mice were sacrificed and the brains were removed. Immunohistochemical staining across the whole brain was performed to detect microglia activation via Iba1 and neuronal activation via c-Fos. Morphology of microglia and the number of c-Fo-positive neurons were analyzed by Image-Pro Premier 3D. One-way ANOVA and Fisher's least-significant differences were employed for statistical analyses. Results Minocycline alleviated LPS-induced neuroinflammation as evidenced by reduced activation of microglia in multiple brain regions, including the shell part of the nucleus accumbens (Acbs), paraventricular nucleus (PVN) of the hypothalamus, central nucleus of the amygdala (CeA), locus coeruleus (LC), and nucleus tractus solitarius (NTS). Minocycline significantly increased the number of c-Fo-positive neurons in NTS and area postrema (AP) after LPS treatment. Furthermore, in NTS-associated brain areas, including LC, lateral parabrachial nucleus (LPB), periaqueductal gray (PAG), dorsal raphe nucleus (DR), amygdala, PVN, and bed nucleus of the stria terminali (BNST), minocycline also significantly increased the number of c-Fo-positive neurons after LPS administration. Conclusion Minocycline alleviates LPS-induced neuroinflammation in multiple brain regions, possibly due to increased activation of neurons in the NTS-associated network.

Animals , Female , Male , Mice , Lipopolysaccharides/toxicity , Mice, Inbred C57BL , Minocycline/pharmacology , Neuroinflammatory Diseases , Solitary Nucleus
Acta Physiologica Sinica ; (6): 901-908, 2021.
Article in Chinese | WPRIM | ID: wpr-921294


The aim of the present study was to investigate the effects of dexmedetomidine (DEX) on acute liver injury induced by lipopolysaccharide (LPS)/D-galactosamine (D-Gal) and the underlying mechanism. Male BALB/c mice were intraperitoneally injected with LPS/D-Gal to induce acute liver injury model, and pretreated with DEX or in combination with the autophagy inhibitor, 3-methyladenine (3-MA) 30 min before injection. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, as well as myeloperoxidase (MPO) activity in liver tissue were determined with the corresponding kits. Serum tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) levels were determined by ELISA. The protein expression levels of LC3-II and P62 in liver tissue were determined by Western blot. Liver histopathological changes were detected by HE staining. The results showed that, compared with control group, LPS/D-Gal enhanced ALT and AST activity, increased TNF-α and IL-6 levels, as well as MPO activity, up-regulated LC3-II and P62 protein expression levels, and significantly induced pathological damage in liver tissue. DEX reversed the above changes in the LPS/D-Gal group, whereas these protective effects of DEX were blocked by 3-MA. The above results suggest that DEX alleviates LPS/D-Gal-induced acute liver injury, which may be associated with the up-regulation of LC3-II protein expression and the activation of autophagy.

Animals , Male , Mice , Alanine Transaminase , Chemical and Drug Induced Liver Injury/drug therapy , Dexmedetomidine/pharmacology , Galactosamine/toxicity , Interleukin-6/blood , Lipopolysaccharides/toxicity , Liver , Mice, Inbred BALB C , Microtubule-Associated Proteins/metabolism , Tumor Necrosis Factor-alpha/blood , Up-Regulation
China Journal of Chinese Materia Medica ; (24): 4800-4807, 2021.
Article in Chinese | WPRIM | ID: wpr-888187


This paper aimed to explore the anti-inflammatory effect of ethanol extract from Saposhnikoviae Radix in a lipopolysaccharide(LPS)-induced inflammation mouse model and its regulation of TLR4/NF-κB signaling pathway. The ethanol extract from Saposhnikoviae Radix was separated and purified on the macroporous adsorption resin and its main chemical components were identified by UPLC-QE/MS. The identification results showed that the top ten components of ethanol extract from Saposhnikoviae Radix were mainly chromones and coumarins. A mouse model of inflammation induced by intraperitoneal injection of LPS was used to investigate the anti-inflammatory effects of ethanol extract from Saposhnikoviae Radix after intragastric administration for seven successive days. Mice in all groups except for the control group were treated with intraperitoneal injection of LPS(0.015 g·kg~(-1)) one hour after the last administration, and twelve hours later, the blood was sampled and separated and the broncoalveolar lavage fluid(BALF) was collected. The levels of nitric oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β) in mouse serum and BALF were detected by ELISA. The harvested lung tissue was stained with hematoxylin-eosin(HE) for observing the pathological changes, followed by the detection of protein expression levels of related molecules in TLR4/NF-κB signaling pathway by Western blotting. The results showed that the ethanol extract from Saposhnikoviae Radix significantly ameliorated the pathological conditions in lung tissue of model mice, reversed the increase in NO, TNF-α, IL-6, and IL-1β levels of mouse serum and BALF, down-regulated the protein expression levels of Toll-like receptor 4(TLR4), myeloid differentiation factor(MyD88), and phosphorylated nuclear transcription factor κB-p65/nuclear transcription factor κB-p65(P-NF-κB p65/NF-κB p65), and up-regulated the NF-κB inhibitory protein α(IκBα). The ethanol extract from Saposhnikoviae Radix exhibited a good anti-inflammatory effect in the LPS-induced acute inflammation muse model, which might be related to the inhibition of the activation of TLR4/NF-κB inflammatory signaling pathway. Chromones and coumarins have been proved to be the active components for its anti-inflammatory effects.

Animals , Mice , Anti-Inflammatory Agents , Ethanol , Inflammation/drug therapy , Lipopolysaccharides/toxicity , NF-kappa B/genetics , Plant Extracts
China Journal of Chinese Materia Medica ; (24): 4774-4781, 2021.
Article in Chinese | WPRIM | ID: wpr-888184


Indolealkylamines(IAAs) are the main hydrophilic substances in toad skin, mainly including free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine. In this study, the LPS-activated neutrophils were used to investigate the structure-activity relationship and anti-inflammatory mechanism of the above-mentioned five monomers from the toad skin in vitro. The neutrophils were divided into the control group, model group(1 μg·mL~(-1) LPS), positive drug group(100 μg·mL~(-1) indometacin), as well as the low-(50 μg·mL~(-1)), medium-(100 μg·mL~(-1)) and high-dose(200 μg·mL~(-1)) free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine groups. The levels of IL-6, TNF-α and IL-1β in the neutrophil supernatant of each group was measured by enzyme-linked immunosorbent assay(ELISA) after LPS stimulation, followed by the detection of apoptosis in each group after Annexin V/PI staining. The protein expression levels of caspase-3, Bax, Bcl-2, beclin1, LC3-I, and LC3-Ⅱ were assayed by Western blot. The results showed that IAAs reduced the excessive secretion of inflammatory cytokines caused by LPS compared with the model group. Besides, the activity of each free IAAs(N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine and dehydrobufotenine), especially bufotenine, was stronger than that of the binding bufothionine. As revealed by Annexin V/PI staining, LPS delayed the early apoptosis of neutrophils compared with the control group, while bufotenine promoted the apoptosis of neutrophils in a dose-dependent manner, which might be related to the elevated expression of apoptosis-related protein Bax/Bcl-2. In addition, LPS activated the autophagy pathways in neutrophils. This study confirmed the efficacy of IAAs in reducing the secretion of inflammatory cytokines in neutrophils induced by LPS for the first time. For instance, bufotenine exerts the anti-inflammatory effect possibly by inducing the apoptosis of neutrophils.

Animals , Anti-Inflammatory Agents/pharmacology , Apoptosis , Bufonidae , Lipopolysaccharides/toxicity , Neutrophils , Skin
Neuroscience Bulletin ; (6): 1671-1682, 2021.
Article in English | WPRIM | ID: wpr-922661


Microglia-mediated neuroinflammation is widely perceived as a contributor to numerous neurological diseases and mental disorders including depression. Discs large homolog 1 (Dlg1), an adaptor protein, regulates cell polarization and the function of K

Animals , Mice , Depression/chemically induced , Inflammation , Lipopolysaccharides/toxicity , Mice, Inbred C57BL , Mice, Knockout , Microglia , NF-kappa B , Neuroinflammatory Diseases
Clinics ; 75: e1665, 2020. tab, graf
Article in English | LILACS | ID: biblio-1133413


OBJECTIVES: This study intended to explore the effect of T regulatory cells (Tregs) in the perinatal liver against LPS-induced inflammation in a preterm birth mouse model. Moreover, the role of adoptive Tregs on the inflammatory response induced by LPS was also studied. METHODS: Female BALB/C mice were injected intraperitoneally (IP) with LPS dissolved in normal saline solution at a dose of 50 µg/kg. Spleens from pregnant mice were used to obtain Tregs. The expression of Forkhead family transcription factor-3 (Foxp3), Interleukin-6 (IL-6), Toll-like receptor-4 (TLR-4), and Heme oxygenase-1 (HO-1) were assessed from fetal liver tissues by polymerase chain reaction and western blotting. RESULTS: LPS administered to mice induced an inflammatory response in the perinatal liver, and this inflammatory response was negatively regulated by Tregs in the experimental group. Maternal-fetal tolerance was maintained by Tregs. Transmission of Tregs was estimated in different experimental groups based on the mRNA expression of TLR-4, IL-6, HO-1, and Foxp3. CONCLUSIONS: After analysis of the experimental data, it was determined that Tregs exhibited regulatory potential against LPS-induced inflammatory response. Further, it was concluded that the transmission of Tregs improved the mother's immune tolerance against LPS-induced inflammation in the fetal liver.

Animals , Female , Pregnancy , Mice , Lipopolysaccharides/toxicity , Premature Birth , T-Lymphocytes, Regulatory , Forkhead Transcription Factors , Inflammation/chemically induced , Liver , Mice, Inbred BALB C
J. appl. oral sci ; 27: e20180291, 2019. graf
Article in English | LILACS, BBO | ID: biblio-984570


Abstract Objective The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). Material and Methods Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. Results In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. Conclusion mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Humans , Male , Female , Adolescent , Teichoic Acids/toxicity , Lipopolysaccharides/toxicity , Enterococcus faecalis/chemistry , Tooth Apex/cytology , Dental Papilla/cytology , Anti-Bacterial Agents/toxicity , Root Canal Irrigants/toxicity , Time Factors , Calcium Hydroxide/toxicity , Calcium Hydroxide/chemistry , Ciprofloxacin/toxicity , Ciprofloxacin/chemistry , Cefaclor/toxicity , Cefaclor/chemistry , Cell Survival/drug effects , Cells, Cultured , Reproducibility of Results , Analysis of Variance , Tooth Apex/drug effects , Dental Papilla/drug effects , Metronidazole/toxicity , Metronidazole/chemistry , Anti-Bacterial Agents
Chinese Critical Care Medicine ; (12): 346-349, 2019.
Article in Chinese | WPRIM | ID: wpr-1010870


OBJECTIVE@#To investigate the protective effect of protein kinase C (PKC) inhibitor rottlerin on rat renal vascular endothelial injury induced by lipopolysaccharide (LPS).@*METHODS@#Rat renal microvascular endothelial cells cultured for 3-6 generations were divided into three groups according to random number table: blank control group in which cells were not challenged, LPS group in which cells were only stimulated by LPS 10 mg/L for 24 hours, and PKC inhibitor group in which cells were treated with PKC inhibitor rottlerin 2 μmol/L 30 minutes before LPS stimulation. The levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-1β, IL-8) were determined by enzyme-linked immunosorbent assay (ELISA). Monolayer permeability was determined by Transwell assay. The expressions of PKC, RhoA and vascular endothelial-cadherin (VE-cadherin) were detected by Western Blot. The morphological characteristic and distribution of F-actin was measured by laser confocal fluorescence microscope.@*RESULTS@#Compared with blank control group, the levels of inflammatory cytokines at 24 hours after 10 mg/L LPS stimulation were significantly increased in LPS group [TNF-α (ng/L): 397.3±25.4 vs. 46.8±8.9, IL-1β (ng/L): 76.7±11.2 vs. 12.6±3.2, IL-8 (ng/L): 574.5±31.4 vs. 73.2±9.6, all P < 0.05], the permeability of endothelial cells was significantly increased (A value: 1.32±0.03 vs. 0.36±0.02, P < 0.05), while the expressions of PKC and RhoA were significantly up-regulated (PKC/β-actin: 0.88±0.02 vs. 0.61±0.03, RhoA/β-actin: 0.96±0.01 vs. 0.49±0.03, both P < 0.05), VE-cadherin expression was significantly down-regulated (VE-cadherin/β-actin: 0.51±0.01 vs. 0.72±0.04, P < 0.05), and the F-actin distribution disorder had obvious stress fiber formation. Compared with LPS group, the levels of inflammatory cytokines were significantly lowered in PKC inhibitor group [TNF-α (ng/L): 127.4±14.6 vs. 397.3±25.4, IL-1β(ng/L): 43.2±7.8 vs. 76.7±11.2, IL-8 (ng/L): 212.7±18.2 vs. 574.5±31.4, all P < 0.05], the permeability of endothelial cells was significantly decreased (A value: 0.81±0.02 vs. 1.32±0.03, P < 0.05), the expressions of PKC and RhoA were significantly down-regulated (PKC/β-actin: 0.44±0.03 vs. 0.88±0.02, RhoA/β-actin: 0.63±0.05 vs. 0.96±0.01, both P < 0.05), the VE-cadherin expression was significantly up-regulated (VE-cadherin/β-actin: 0.69±0.03 vs. 0.51±0.01, P < 0.05), and the F-actin remodeling and stress fiber formation were significantly reduced.@*CONCLUSIONS@#PKC inhibitor could significantly attenuate the damage of vascular endothelial barrier induced by LPS, and plays an important role in endothelial cell barrier.

Animals , Rats , Acute Kidney Injury/prevention & control , Endothelium, Vascular/drug effects , Interleukin-1beta , Lipopolysaccharides/toxicity , Protein Kinase C/antagonists & inhibitors , Protein Kinase Inhibitors/pharmacology , Random Allocation
Braz. oral res. (Online) ; 31: e17, 2017. tab, graf
Article in English | LILACS | ID: biblio-839523


Abstract Periodontitis develops as a result of a continuous interaction between host cells and subgingival pathogenic bacteria. The periodontium has a limited capacity for regeneration, probably due to changes in periodontal ligament stem cells (PDLSCs) phenotype. The aim of this study was to evaluate the effects of lipopolysaccharides from Porphyromonas gingivalis (PgLPS) on mesenchymal phenotype and osteoblast/cementoblast (O/C) potential of PDLSCs. PDLSCs were assessed for Toll-like receptor 2 (TLR2) expression by immunostaining technique. After, cells were exposed to PgLPS, and the following assays were carried out: (i) cell metabolic activity using MTS; (ii) gene expression for IL-1β, TNF-α and OCT-4 by real-time polymerase chain reaction (RT-qPCR); (iii) flow cytometry for STRO-1 and CD105, and (iv) osteogenic differentiation. PDLSCs were positive for TLR2. PgLPS promoted cell proliferation, produced IL-1β and TNF-α, and did not affect the expression of stem cell markers, STRO-1, CD105 and OCT-4. Under osteogenic condition, PDLSCs exposed to PgLPS showed a similar potential to differentiate toward osteoblast/cementoblast phenotype compared to control group as revealed by mineralized matrix deposition and levels of transcripts for RUNX2, ALP and OCN. These results provide evidence that PgLPS induces pro-inflammatory cytokines, but does not change the mesenchymal phenotype and osteoblast/cementoblast differentiation potential of PDLSCs.

Humans , Osteogenesis/drug effects , Periodontal Ligament/cytology , Lipopolysaccharides/toxicity , Porphyromonas gingivalis , Mesenchymal Stem Cells/drug effects , Time Factors , Gene Expression , Osteocalcin/analysis , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Tumor Necrosis Factor-alpha/analysis , Statistics, Nonparametric , Cell Proliferation/drug effects , Alkaline Phosphatase/analysis , Octamer Transcription Factor-3/analysis , Toll-Like Receptors/analysis , Core Binding Factor Alpha 1 Subunit/analysis , Interleukin-1beta/analysis , Mesenchymal Stem Cells/metabolism , Real-Time Polymerase Chain Reaction , Flow Cytometry
Rev. bras. epidemiol ; 18(supl.2): 146-157, Out.-Dez. 2015. tab, graf
Article in English | LILACS | ID: lil-776710


RESUMO: Objetivo: Descrever o perfil dos pacientes que referiram diagnóstico médico de câncer e descrever os tipos de câncer mais prevalentes, segundo variáveis selecionadas. Métodos: Estudo descritivo que utilizou dados da Pesquisa Nacional de Saúde (PSN) de 2013 para estimar prevalências e respectivos valores do intervalo de confiança (IC95%). Resultados: Menos de 2% dos adultos referiram diagnóstico médico de câncer, sendo mais relatado por mulheres, por pessoas com mais de 60 anos, entre brancos, em residentes da zona urbana e moradores da Região Sul. O câncer de próstata foi o mais referido entre os homens; entre as mulheres, o câncer de mama foi o mais prevalente. A menor idade média do primeiro diagnóstico foi identificada para câncer de colo de útero (35,4 anos; IC95% 30,3 - 40,6), e a maior, para câncer de próstata (65,7 anos; IC95% 64,2 - 67,0). Conclusão: Os achados deste estudo são importantes para o planejamento dos serviços de saúde e do seu acesso, pois revelam diferenças, principalmente regionais.

ABSTRACT: Objective: To describe the profile of patients who reported a medical diagnosis of cancer and describe the most prevalent types of cancer, according to selected variables. Methods: A descriptive study that used data from the National Survey of Health, 2013, to estimate prevalence and their values of confidence interval (95%CI). Results: Less than 2% of adults reported a medical diagnosis of cancer, with most reported by women, people over 60, among whites, residents in the village and residents of South Prostate cancer was the most reported among men and breast among women. The lowest average age of first diagnosis was identified for cervical cancer (35.4 years; 95%CI 30.3 - 40.6) and the highest for prostate (65.7 years; 95%CI 64.2 - 67.0). Conclusion: The findings of this study are important for the planning of health services and access, as they show differences mainly regional.

Animals , Mice , Anti-Inflammatory Agents/therapeutic use , Curcumin/therapeutic use , Drug Carriers , Interleukin-1beta/genetics , Lipids/chemistry , Lipopolysaccharides/toxicity , Nanoparticles , Sepsis/drug therapy , Cytokines/blood , Inflammation Mediators/metabolism , Mice, Transgenic , Promoter Regions, Genetic , Signal Transduction , Sepsis/chemically induced
Arq. neuropsiquiatr ; 73(9): 784-790, Sept. 2015. ilus
Article in English | LILACS | ID: lil-757393


Inflammation and oxidative stress have important roles in memory impairment. The effect of 7-nitroindazole (7NI) on lipopolysaccharide (LPS)-induced memory impairment was investigated. Rats were used, divided into four groups that were treated as follows: (1) control (saline); (2) LPS; (3) 7NI-LPS; and (4) 7NI before passive avoidance (PA). In the LPS group, the latency for entering the dark compartment was shorter than in the controls (p < 0.01 and p < 0.001); while in the 7NI-LPS group, it was longer than in the LPS group (p < 0.01 and p < 0.001). Malondialdehyde (MDA) and nitric oxide (NO) metabolite concentrations in the brain tissues of the LPS group were higher than in the controls (p < 0.001 and p < 0.05); while in the 7NI-LPS group, they were lower than in the LPS group (p < 0.001 and p < 0.05, respectively). The thiol content in the brain of the LPS group was lower than in the controls (p < 0.001); while in the 7NI-LPS group, it was higher than in the LPS group (p < 0.001). It is suggested that brain tissue oxidative damage and NO elevation have a role in the deleterious effects of LPS on memory retention that are preventable using 7NI.

Inflamação e estresse oxidativo tem importante papel no déficit de memória. O efeito do 7-nitroindazol (7NI) no déficit de memória induzido por lipossacarídeos (LPS) foi investigado. Foram utilizados ratos que foram divididos em quatro grupos e tratados da seguinte maneira: (1) controles (solução salina); (2) LPS; (3) 7NI-LPS; e (4) 7NI antes da esquiva passiva (PA). No grupo LPS, a latência para entrar no compartimento escuro foi mais curta que nos controles (p < 0,01 e p < 0,001); enquanto no grupo 7NI-LPS, a latência foi maior que aquela do grupo LPS (p < 0,01 e p < 0,001). Concentrações de malondialdeído (MDA) e metabólitos do ácido nítrico (NO) no tecido cerebral do grupo LPS foram maiores que aquelas dos controles (p < 0,001 e p < 0,05); enquanto no grupo 7NI-LPS, as concentrações foram menores do que no grupo LPS (p < 0,001 e p < 0,05, respectivamente). O conteúdo cerebral de tiol no grupo LPS foi menos do que nos controles (p < 0,001); enquanto no grupo 7NI-LPS, este conteúdo foi maior que no grupo LPS (p < 0,001). Sugere-se que o dano oxidativo cerebral e o aumento de NO tenham um papel nos efeitos deteriorativos dos LPS na memória de retenção, e que isto possa ser prevenido com o uso de 7NI.

Animals , Male , Rats , Indazoles/pharmacology , Lipopolysaccharides/toxicity , Memory Disorders/prevention & control , Memory/drug effects , Neuroprotective Agents/pharmacology , Brain Chemistry , Memory Disorders/chemically induced , Rats, Wistar
Dental press j. orthod. (Impr.) ; 20(2): 68-75, Mar-Apr/2015. tab, graf
Article in English | LILACS | ID: lil-745863


OBJECTIVE: The aim of this cross sectional study was to assess serum insulin-like growth factor-1 (IGF-1) levels in female and male subjects at various cervical vertebral maturation (CVM) stages. MATERIAL AND METHODS: The study sample consisted of 60 subjects, 30 females and 30 males, in the age range of 8-23 years. For all subjects, serum IGF-1 level was estimated from blood samples by means of chemiluminescence immunoassay (CLIA). CVM was assessed on lateral cephalograms using the method described by Baccetti. Serum IGF-1 level and cervical staging data of 30 female subjects were included and taken from records of a previous study. Data were analyzed by Kruska-Wallis and Mann Whitney test. Bonferroni correction was carried out and alpha value was set at 0.003. RESULTS: Peak value of serum IGF-1 was observed in cervical stages CS3 in females and CS4 in males. Differences between males and females were observed in mean values of IGF-1 at stages CS3, 4 and 5. The highest mean IGF-1 levels in males was observed in CS4 followed by CS5 and third highest in CS3; whereas in females the highest mean IGF-1 levelswas observed in CS3 followed by CS4 and third highest in CS5. Trends of IGF-1 in relation to the cervical stages also differed between males and females. The greatest mean serum IGF-1 value for both sexes was comparable, for females (397 ng/ml) values were slightly higher than in males (394.8 ng/ml). CONCLUSIONS: Males and females showed differences in IGF-1 trends and levels at different cervical stages. .

OBJETIVO: o objetivo do presente estudo transversal foi avaliar os níveis do fator de crescimento semelhante à insulina-1 (IGF-1 sérico) em pacientes de ambos os sexos e em diferentes estágios de maturação das vértebras cervicais (MVC). MÉTODOS: a amostra consistiu de 60 pacientes, sendo 30 do sexo masculino e 30 do sexo feminino, com idades entre 8 e 23 anos. Amostras de sangue foram colhidas de todos os pacientes, cujos níveis de IGF-1 sérico foram avaliados por meio do método de imunoensaio quimioluminescente (CLIA). O estágio de MVC foi avaliado por meio de radiografias cefalométricas de perfil por meio do método descrito por Baccetti. O nível de IGF-1 sérico e o estágio de maturação das vertebras cervicais de 30 pacientes do sexo feminino foram avaliados e os dados retirados dos registros de um estudo prévio. Os dados foram submetidos aos testes de Kruskal-Wallis e de Mann-Whitney. A correção de Bonferroni foi calculada e o valor de alfa foi de 0,003. RESULTADOS: o valor de pico do IGF-1 sérico foi encontrado no estágio CS3, para mulheres, e CS4, para homens. Foram encontradas diferenças entre as médias dos valores de IGF-1 entre homens e mulheres nos estágios CS3, 4 e 5. O valor médio mais alto para os níveis de IGF-1 nos homens foi observado no estágio CS4, seguido do estágio CS5 e CS3. Nas mulheres, o valor médio mais alto foi observado em CS3, seguido do estágio CS4 e CS5. Diferenças também foram encontradas quanto à curva do IGF-1, em relação ao estágio de maturação das vértebras cervicais nos pacientes de ambos os sexos. O valor médio de IGF-1 sérico mais alto foi comparado. As pacientes do sexo feminino apresentaram valores ligeiramente mais altos (397ng/ml) em comparação aos pacientes do sexo masculino (394.8ng/ml). CONCLUSÕES: homens e mulheres apresentam valores de IGF-1 diferentes em estágios de maturação das vértebras cervicais diferentes. .

Animals , Mice , Endoplasmic Reticulum/metabolism , Inflammation Mediators/metabolism , Macrolides/metabolism , Mycobacterium ulcerans/pathogenicity , Buruli Ulcer/metabolism , Buruli Ulcer/microbiology , Buruli Ulcer/pathology , Cell Line , Cell Adhesion Molecules , Endoplasmic Reticulum/pathology , Lipopolysaccharides/toxicity , Mycobacterium ulcerans/metabolism , Protein Biosynthesis/drug effects , Protein Transport/drug effects , Tumor Necrosis Factor-alpha
Int. j. morphol ; 33(1): 194-203, Mar. 2015. ilus
Article in English | LILACS | ID: lil-743785


The present study was to investigate the effects of visfatin on the morphological structure and function of the rat uterus during inflammation. The expression and distribution of visfatin, morphological structure, eosinophils (EOS), myeloperoxidase (MPO) and cytokines in the uterus of the LPS-induced rat were studied using hematoxylin-eosin staining (HE), immunohistochemical methods, western blots and enzyme-linked immunosorbent assay (ELISA). The present study showed that visfatin positive cells dispersed widely in the uterus, and strong positive staining was observed mainly in the cell cytoplasm. Compared with saline group, in visfatin group, more uterine glands were found, EOS increased, and the difference was significant (P<0.05), MPO reduced, and the difference was significant (P<0.01). In addition, visfatin was able to increase the secretion of IL-1b, IL-6, and TNF-a (P<0.01). Compared with LPS group, in vifatin+LPS group, the uterine glands of the lamina propria increased, the myometrium became thinner, the number of EOS and MPO reduced obviously, but the difference was not significant (P>0.05), and after LPS stimulated body, visfatin decrease the level of IL-1b, IL-6, TNF-a (P<0.01). The above results suggest that visfatin could affect the morphological structure of rat uterus; Visfatin could modulate the inflammatory response in rats' uterus by regulating the quantity of inflammatory cells, such as EOS and MPO, and the level of inflammatory cytokines, such as IL-1b, IL-6, TNF-a.

El objetivo del presente estudio fue investigar los efectos de la visfatina sobre la estructura morfológica y la función del útero de la rata durante la inflamación. Se estudiaron la expresión y distribución de la visfatina, la estructura morfológica, eosinófilos, mieloperoxidasa y citoquinas en el útero de rata mediante la tinción de H&E, métodos inmunohistoquímicos, Western blots y ELISA. El estudio mostró que las células visfatina positivas se dispersan ampliamente en el útero, junto a una fuerte tinción positiva, principalmente en el citoplasma de la célula. En comparación con el grupo control, en el grupo visfatina, se encontraron más glándulas uterinas, se observó un aumento de EOS y la diferencia fue significativa (p<0,05), MPO reducida siendo esta diferencia también significativa (p<0,01). Además, la visfatina fue capaz de aumentar la secreción de IL-1b, IL-6 y TNF-a (P<0,01). En comparación con el grupo LPS, visfatina+grupo LPS, las glándulas uterinas de la lámina propia aumentaron, se observó un miometrio más delgado, y número reducido de EOS y MPO, sin embargo, la diferencia no fue significativa (P>0,05). Después de estímulo LPS en el cuerpo, se registró un nivel menor de visfatina en IL-1b, IL-6, TNF-a (P<0,01). Los resultados anteriores sugieren que visfatina podría afectar a la estructura morfológica del útero de rata. Además, podría modular la respuesta inflamatoria en el útero mediante la regulación de la cantidad de células inflamatorias, tales como EOS y MPO.

Animals , Female , Rats , Uterus/drug effects , Lipopolysaccharides/toxicity , Nicotinamide Phosphoribosyltransferase/pharmacology , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Blotting, Western , Rats, Wistar , Peroxidase/drug effects , Inflammation , Neutrophils/drug effects
Rev. bras. enferm ; 67(5): 722-729, Sep-Oct/2014. tab
Article in Portuguese | LILACS, BDENF | ID: lil-731212


Objetivou-se estimar o tempo de decisão para procura de atendimento (TD) para homens e mulheres com infarto agudo do miocárdio (IAM); analisar a influência de variáveis ambientais no TD e a interação entre gênero e variáveis ambientais para o desfecho TD. Estudo transversal, envolvendo cem pacientes, entrevistados em hospitais de Salvador. Na análise dos dados empregou-se o Qui-quadrado ou Exato de Fisher e o Modelo de Regressão Linear Robusto. Predominou o IAM ocorrido no domicílio, familiares no entorno, e os pacientes sendo alvo de ações equivocadas. Observou-se TD elevado para mulheres (0,9h) e homens (1,4h). Aqueles em casa no início dos sintomas tiveram maior TD, comparados aos no trabalho e menor em relação aos em via pública (p=0,047). Houve interação estatisticamente significante entre gênero e viver acompanhado; e entre gênero e ter companheiro e filhos no entorno, para o desfecho TD. O cuidar em enfermagem focalizado nas especificidades de fatores ambientais e de gênero pode otimizar o atendimento precoce.

The purpose was to estimate the decision time (DT) for searching for attendance for men and women suffering from acute myocardial infarction (AMI); and to analyze the influence of surrounding variables in the DT. Transversal study, involving one hundred patients interviewed in hospitals of Salvador-BA, Brazil. For data analysis, it was used the chi-squared or Fisher’s exact test, and the Robust Linear Regression Model. AMI at the home predominated, with family members and patients receiving mistaken actions. A high DT was observed both, for women (0.9h) and men (1.4h). Those at home during the initial symptoms had higher DT, compared to those at work; and lower in relation to those in public spaces (p=0.047). Statistically significant interaction occurred among gender and the fact of living with company; and among gender and having a companion and children, for the outcome of the DT. Nursing care focused on the specificity of surrounding factors and gender can optimize early attendance.

Se objetivó estimar el tiempo de decisión para buscar atendimiento (TD) para hombres y mujeres con infarto agudo de miocardio (IAM); analizar la influencia de variables ambientales en TD y la interacción entre genero y variables ambientales para el desfecho del TD. Estudio transversal, envolviendo cien pacientes entrevistados en hospitales de Salvador-BA, Brasil. En el análisis se utilizó el chi-cuadrado o el Teste Exacto de Fisher y el Modelo de Regresión Linear Robusto. Predominó el IAM en el domicilio, familiares en el entorno y con pacientes siendo objeto de acciones equivocadas. Se observó TD elevados para mujeres (0,9h) y hombres (1,4h). Aquellos en sus casas en el inicio de los síntomas tuvieron mayor TD, comparados a los en el trabajo, y menor en relación aquellos en vía pública (p=0,047). Hubo interacción estadísticamente significante entre genero y vivir acompañado y entre genero y tener compañero e hijos en el entorno, para el desfecho del TD. El cuidar en enfermería focalizado en especificidades de factores ambientales y de géneros puede optimizar el atendimiento precoce.

Animals , Male , Rats , Growth Hormone/pharmacology , /pharmacology , Lipopolysaccharides/toxicity , Repressor Proteins , Transcription Factors , Tumor Necrosis Factor-alpha/pharmacology , Drug Resistance , Insulin-Like Growth Factor I/genetics , Proteins/genetics , Rats, Sprague-Dawley , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Somatotropin/genetics , Suppressor of Cytokine Signaling Proteins
Rev. bras. enferm ; 67(5): 715-721, Sep-Oct/2014. tab
Article in Portuguese | LILACS, BDENF | ID: lil-731216


Objetivou-se validar as diretrizes gerais da comunicação do enfermeiro com o cego. Estudo quantitativo, realizado entre abril de 2008 e março de 2009 em Fortaleza-CE. Participaram 30 enfermeiros e 30 cegos divididos em grupo controle e experimental. Realizaram-se 30 consultas de enfermagem filmadas e analisadas por especialistas. O grupo experimental apresentou desempenho bom e excelente em todas as diretrizes para a comunicação verbal e não verbal com o cego, contrariamente ao grupo controle. Os resultados do estudo apontam para a urgência da adoção do ensino destas diretrizes gerais de comunicação com cegos nos cursos de enfermagem, além de capacitar enfermeiros no cuidado a pessoas cegas.

This quantitative study, conducted between 2008 April and 2009 March in Fortaleza-CE, Brazil, aimed to validate the general guidelines of the communication of the nurse with the blind. Thirty nurses and 30 blinds, divided into control and experimental groups, participated in the study. Thirty nursing consultations were videotaped and analyzed by experts. In contrast to the control group, the experimental group showed good and excellent performance in all guidelines for verbal and non-verbal communication with the blinds. The study results point to the urgency of adopting the teaching of these general guidelines for communicating with the blind in nursing courses, in addition to training nurses in caring for the blind people.

Estudio cuantitativo, realizado entre abril de 2008 y marzo de 2009, en Fortaleza-CE, Brasil, que tuvo como objetivo validar los lineamientos generales de la comunicación del enfermero con los ciegos. Los participantes fueron 30 enfermeros y 30 ciegos, divididos en grupos control y experimental. Fueran realizadas 30 consultas de enfermería, registradas y analizadas por expertos. El grupo experimental mostró un buen y excelente rendimiento en todas las directrices para comunicación verbal y no verbal con los ciegos, en contraste con el grupo de control. Los resultados del estudio apuntan a la urgencia de la adopción de la enseñanza de estas directrices generales para la comunicación con los ciegos en los cursos de enfermería, además de la formación de enfermeras en el cuidado de las personas ciegas.

Animals , Female , Rats , Hypertension, Portal/etiology , Lipopolysaccharides/toxicity , Portal System/drug effects , Disease Models, Animal , Hemodynamics/drug effects , Hypertension, Portal/physiopathology , Portal System/physiology , Rats, Sprague-Dawley
Braz. j. med. biol. res ; 47(3): 231-236, 03/2014. graf
Article in English | LILACS | ID: lil-704623


Studies have shown that edaravone may prevent liver injury. This study aimed to investigate the effects of edaravone on the liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in female BALB/c mice. Edaravone was injected into mice 30 min before and 4 h after GalN/LPS injection. The survival rate was determined within the first 24 h. Animals were killed 8 h after GalN/LPS injection, and liver injury was biochemically and histologically assessed. Hepatocyte apoptosis was measured by TUNEL staining; proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in the liver were assayed by ELISA; expression of caspase-8 and caspase-3 proteins was detected by Western blot assay; and caspase-3 activity was also determined. Results showed that GalN/LPS induced marked elevations in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Edaravone significantly inhibited elevation of serum AST and ALT, accompanied by an improvement in histological findings. Edaravone lowered the levels of TNF-α and IL-6 and reduced the number of TUNEL-positive cells. In addition, 24 h after edaravone treatment, caspase-3 activity and mortality were reduced. Edaravone may effectively ameliorate GalN/LPS-induced liver injury in mice by reducing proinflammatory cytokines and inhibiting apoptosis.

Animals , Female , Antipyrine/analogs & derivatives , Apoptosis/drug effects , Cytokines/drug effects , Chemical and Drug Induced Liver Injury/prevention & control , Free Radical Scavengers/pharmacology , Alanine Transaminase/blood , Antipyrine/pharmacology , Aspartate Aminotransferases/blood , /analysis , /metabolism , /analysis , Chemical and Drug Induced Liver Injury/physiopathology , Enzyme-Linked Immunosorbent Assay , Endotoxins/toxicity , Galactosamine/toxicity , Hepatocytes/drug effects , In Situ Nick-End Labeling , /analysis , Lipopolysaccharides/toxicity , Mice, Inbred BALB C , Random Allocation , Tumor Necrosis Factor-alpha/analysis