Memantine, A NMDA Receptor Inhibitor Attenuate Lipopolysaccharide-Induced Lung Inflammation and Oxidative Damage in Mice.

Objective To investigate the effects and potential mechanisms of somatostatin (SST)against oxidative stress in a mouse model of lipopolysaccharide (LPS)-induced acute lung injury (ALI).Methods A total of 96 ICR male mice of specific pathogen free (SPF) were divided randomly (random number) into four groups:normal control group (n =24),SST control group (n =24),ALI group (n =24) and SST treated group (n =24).The mice of ALI group were given the intra-peritoneal injection of LPS in dose of 12 mg in solution of 40 mL/kg.Mice of normal control group were given the intra-peritoneal injection of equivalent volume of normal saline (NS 40 mL/kg) instead,and mice of SST control group were given the intra-peritoneal injection of equivalent volume of normal saline (40 mL/kg) instead,then hypodermic injection of SST (20 μg:20 mL/kg) at 0.5,2,6 or 12 h after normal saline administration,and mice of SST treated group were given the intra-peritoneal injection of LPS (12 mg:40 mL/kg),then hypodermic injection of SST (20 μg:20 mL/kg) at 0.5,2,6 or 12 h after LPS administration.All mice were sacrificed at 3,8 h or 16 h after the first injection of LPS or NS.The lung wet/dry ratios were calculated.Pathological changes of lung tissues were observed under light microscope after HE staining.The levels of malondialdehyde (MDA),superoxide dismutase (SOD) and catalase (CAT) in the lung tissues were determined by chemical colorimetry; quinone oxidoreductase (NQO-1) levels in the lung tissues were measured by ELISA ; the expressions of nuclear factor-E2-related factor2 (Nrf2) and its protein levels in the lung tissues were measured by RT-PCR and Western blotting,respectively.One-way analysis of variance (ANOVA) was employed for statistical analysis by using SPSS version 19.0 to compare values among all groups.Results There were no significant differences in all biomarker variables between normal control group and SST control group (P ＜ 0.05).Lung wet/dry ratios in ALI group at 8 h and 16 h were significantly higher than those in normal control group (P =0.000) and obviously lower in SST treated group (5.21 ±0.13) vs.(5.78±0.20),(5.39±0.29) vs.(6.17 ±0.17) (P=0.000).Compared with normal control group,the damage of lung tissues in ALI group was more severe.In contrast,histopathological lesions observed in SST treated group were milder significantly.The MDA and NQO-1 in the lung tissues were much higher in ALI group than those in normal control group,while the activities of SOD and CAT were much lower in ALI group (P =0.000).Compared with ALI group,the MDA level (ng/mg) in SST treated group decreased remarkably (2.66 ±0.18) vs.(3.58 ±0.26),(3.52 ±0.31)vs.(4.37 ±0.19),(3.81 ±0.38) vs.(4.92 ±0.25) (P=0.000),whereas CAT activity (U/mg)increased significantly (9.21± 0.47) vs.(7.90±0.32),(10.06±0.51) vs.(8.39±0.41),(10.98 ± 0.33) vs.(9.52 ± 0.55) (P =0.000) ; SOD activity and NQO-1 level at 3 h had no obvious increase (P =0.359、0.111),but SOD activities and NQO-1 levels were markedly increased at 8 h and 16 h [SOD (U/mg):(75.34±5.5) vs.(67.89±3.8),P=0.021; (64.82±4.2) vs.(50.31 ±5.0),P=0.000],[NQO-1 (ng/mg):(1.052 ±0.041) vs.(0.715±0.038),(1.338±0.027)vs.(0.532 ± 0.028),P =0.000].Compared with normal control group,the Nrf2 mRNA and protein levels in ALI group were higher markedly (P =0.000),and Nrf2 mRNA in SST treated group were much higher (P =0.023-0.000),while Nrf2 protein level at 3 h showed no significant difference (P =0.101)but increased obviously at 8 h,16 h (P =0.000).Conclusions Our study has shown that SST can upregulate Nrf2 signal pathway,which may have the effects on regulating the immune response and protections against oxidative damages in ALI. Key words: Lipopolysaccharide ; Acute lung injury; Somatostatin ; Oxidative Stress ; Nuclear factor-E2-related factor 2 ; Mice

Cedrol ameliorates lipopolysaccharide-induced systemic inflammation and lung injury in rats.

Objective To investigate the inhibitory effect of kukoamine B (KB) on high mobility group protein B1 (HMGB1)/nuclear factor-κB (NF-κB) signaling pathway in lung injury of induced septic mice and its protective effect. Methods Thirty C57BL/6 mice were randomly divided into three groups (n = 10): control group, lipopolysaccharide (LPS) model group and KB intervention group. Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg LPS, while equivalent normal saline was given in control group, and 20 μg/kg KB was injected through caudal vein 4 hours after LPS challenge in KB intervention group. The blood and lung tissue samples were harvested 24 hours after LPS injection. The lung wet/dry weight ratio (W/D), the activity of myeloperoxidase (MPO) and protein content in bronchoalveolar lavage fluid (BALF) were determined, and tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in plasma and lung tissue were determined by enzyme linked immunosorbent assay (ELISA). Western Blot was applied for determining the protein expressions of HMGB1 and NF-κB. Results Compared with control group, LPS could significantly increase lung W/D ratio, MPO activity, protein content in BALF, the levels of TNF-α and IL-1β in plasma and lung tissues, and protein expressions of HMGB1 and NF-κB [lung W/D ratio: 6.84±0.42 vs. 3.36±0.31, lung MPO (U/g): 36.46±2.72 vs. 2.32±0.52, protein content in BALF (g/L): 6.21±1.35 vs. 0.35±0.12, plasma TNF-α (ng/L): 36.31±1.53 vs. 8.53±0.82, lung TNF-α (ng/L): 65.35±2.32 vs. 12.36±1.47, plasma IL-1β (ng/L): 64.26±7.53 vs. 28.83±5.74, lung IL-1β (ng/L): 542.36±14.47 vs. 58.46±7.24, lung HMGB1 (gray value): 316.43±4.26 vs. 100.21±0.06, lung NF-κB (gray value): 465.84±6.38 vs. 100.15±0.08, all P < 0.05], which indicated that LPS induced mice sepsis model was successful, and lung injury occurred. However, compared with the mice in LPS group, KB could significantly provide the protective effects for lung injury, and alleviate the changes in above parameters [lung W/D ratio: 4.56±0.43 vs. 6.84±0.42, lung MPO activity (U/g): 14.83±1.47 vs. 36.46±2.72, protein content in BALF(g/L): 2.33±1.24 vs. 6.21±1.35, plasma TNF-α (ng/L): 12.53±0.42 vs. 36.31±1.53, lung TNF-α (ng/L): 32.27±1.41 vs. 65.35±2.32, plasma IL-1β (ng/L): 42.37±6.24 vs. 64.26±7.53, lung IL-1β (ng/L): 314.42±17.38 vs. 542.36±14.47, lung HMGB1 (gray value): 224.57±3.42 vs. 316.43±4.26, lung NF-κB (gray value): 258.73±5.42 vs. 465.84±6.38, all P < 0.05], which indicated that KB had obvious protective effect on the lung of septic mice. Conclusion KB could provide protective effect on lung injury in LPS-induced septic mice through anti-inflammation, which is related to HMGB1/NF-κB signaling pathway. Key words: Sepsis; Kukoamine B; High mobility group protein B1; Nuclear factor-κB; Inflammatory reaction; Lung

Here, we investigated the protective efficacy of protocatechuic acid (PCA) against lipopolysaccharide (LPS)-induced septic lung injury. Eighty-two male Balb/c mice were divided into six groups: control, PCA30 (30mg/kg), LPS (10mg/kg), PCA10-LPS, PCA20-LPS, and PCA30-LPS treated with 10, 20 and 30mg/kg PCA, respectively, for seven days before intraperitoneal LPS injection. PCA pre-treatment, especially at higher dose, significantly reduced LPS-induced lung tissue injury as indicated by increased heat shock protein 70 and antioxidant molecules (reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) accompanied by lower oxidative stress indices (malondialdehyde and nitric oxide). PCA administration decreased inflammatory mediators including myeloperoxidase, nuclear factor kappa B (NF-κB p65), and pro-inflammatory cytokines, and prevented the development of apoptotic events in the lung tissue. At the molecular level, PCA downregulated mRNA expression of nitric oxide synthase 2, C/EBP homologous protein, and high mobility group box1 in the lungs of all PCA-LPS treated mice. Thus, PCA-pre-treatment effectively counteracted sepsis-induced acute lung injury in vivo by promoting and antioxidant status, while inhibiting inflammation and apoptosis. PRACTICAL IMPLICATIONS: Sepsis-mediated organ dysfunction and high mortality is aggravated by acute lung injury (ALI). Therefore, new therapeutic approaches are needed to encounter sepsis-mediated ALI. Protocatechuic acid (PCA) is a naturally occurring phenolic acid with various biological and pharmacological activities. PCA is abundant in edible plants including Allium cepa L., Oryza sativa L., Hibiscus sabdariffa, Prunus domesticaL., and Eucommia ulmoides. In this investigation we studied the potential protective role of pure PCA (10, 20 and 30mg/kg) on LPS-mediated septic lung injury in mice through examining oxidative challenge, inflammatory response, apoptotic events and histopathological changes in addition to evaluating the levels and mRNA expression of heat shock protein 70, C/EBP homologous protein and high mobility group box1 in the lung tissue. The recorded results showed that PCA pre-administration was able to significantly abrogate the damages in the lung tissue associated septic response. This protective effect comes from its strong antioxidant, anti-inflammatory, and anti-apoptotic activities, suggesting that PCA may be applied to alleviate ALI associated with the development of sepsis.

Objective To investigate the effects of interlukin-10 (IL-10) on lipopolysaccharide (LPS)-induced liver injury in rats.Methods One hundred male Wistar rats,aged 10-14 weeks,weighing 250-300 g,were randomly divided into 5 groups (n =20 each):control group (C group),LPS group,IL-10 group,HO-1 inducer cobalt protoporphyrin-Ⅸ group (Co group) and HO-1 inhibitor zinc protoporphyrin-Ⅸ group (Zn group).The animals in LPS,IL-10,Co and Zn groups received intraperitoneal LPS 20 mg/kg.IL-10,Co and Zn groups received recombinant human IL-10 1 μg at 3 h before LPS injection.Co and Zn groups received cobalt protoporphyrin-Ⅸ and zinc protoporphyrin-Ⅸ 25 mg/kg at 2 h before administration of recombinant human IL-10.Ten rats in each group were chosen at 24 h after LPS injection and blood samples were collected from the heart for determination of the levels of serum alanine aminotransferase (ALT),aspartate transaminase (AST),tumor necrosis factor-α (TNF-α) and interlukin-1β (IL-1 β).The animals were then sacrificed and lungs removed for determination of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities,malonodialdehyde (MDA) content and expression of HO-1 mRNA in lung tissues.The left 10 rats in each group were chosen and the survival rates within 72 h after LPS administration were recorded.Results Compared with C group,the levels of serum ALT,AST,TNF-αand IL-1 β and MDA content in lung tissues were significantly increased,and GSH-Px and SOD activities in lung tissues and survival rates were decreased in LPS,IL-10,Co and Zn groups (P ＜ 0.05).Compared with LPS group,the levels of serum ALT,AST,TNF-α,IL-1β and MDA content in lung tissues were significantly decreased,GSH-Px and SOD activities in lung tissues and survival rates were increased,and the expression of HO-1 mRNA was up-regulated (P ＜ 0.05).Compared with IL-10 group,the levels of serum ALT,AST,TNF-α and IL-1β and MDA content in lung tissues were significantly increased,and GSH-Px and SOD activities in lung tissues and survival rates were decreased in group Zn (P ＜ 0.05),and no significant change in the parameters mentioned above was found in Co group (P ＞ 0.05).Conclusion IL-10 can attenuate LPS-induced liver injury in rats by inducing the expression of HO-1. Key words: Interlukin-10 ; Endotoxemia; Liver

Objective: To investigate whether glabridin has a beneficial effect on lipopolysaccharide (LPS) induced acute respiratory distress syndrome (ARDS) in rats, and to explore the possible underlying mechanisms. Methods: Thirty-two Wistar rats were randomly assigned into control group, model group (LPS group), glabridin group (GLA group), and ulinastatin group (UTI group), with 8 rats in each group. ARDS rat model was reproduced by intraperitoneal injection of LPS (10 mg/kg). The rats in the control group received an equal volume of normal saline at the same times. The rats in GLA group were gavaged by glabridin (30 mg/kg). The rats in UTI group were injected ulinastatin (20 000 U/kg). Animals were sacrificed 12 hours after LPS challenge. Plasma and lung tissue samples were collected. Histopathological evaluation, lung wet/dry (W/D)ratio, tumor necrosis factor-α (TNF-α), interleukin-18 (IL-18), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase(SOD)were analyzed. Immunohistochemical method was used to detect the protein expression of p38MAPK and ERK. Western blot method was used to detect the changes of p38 mitogen activated protein kinase (p-p38MAPK) and phosphorylated extracellular regulated protein kinases (pERK) protein expression in lung tissues. Result: In the control groups, lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group, ARDS characters such as extensive thickening of the alveolar wall, significant infiltration of inflammatory cells, demolished structure of pulmonary alveoli, and hemorrhage were found. In the GLA and UTI treatment group, these pathological changes in lung were markedly alleviated compare with LPS-induced ARDS group. Compared with control groups, lung W/D ratio, TNF-α and IL-18 in plasma, and lung MDA, NO levels in lung homogenates of the LPS group were increased significantly, while the lung SOD levels of the LPS group were decreased. Compared with the LPS group, lung W/D ratio, TNF-α and IL-18 in plasma , and lung MDA, NO levels in lung homogenates of the GLA group and UTI group were decreased significantly, while the lung SOD levels of the GLA and ulinastatin groups were increased [TNF-α(μg/L): 51.7±10.3 vs 105.7±30.5, IL-18(μg/L): 37.9±13.9 vs 49.2±14.5, MDA (nmol/mgprot): 2.87±0.62 vs 3.81±0.42, NO(μmol/L): 18.96±0.79 vs 28.58±2.51, SOD(U/mgprot): 115.5±15.2 vs 75.9±14.0, all P<0.05]. Immunohistochemistry showed that the positive expressions of p38MAPK and ERK in cytoplasm and nucleus of the glabridin and ulinastatin treatment group were significantly lower than those of the model group. Western blot showed that compared with the control group, the p-p38MAPK and pERK protein expression in LPS group were significantly increased. And the glabridin and ulinastatin inhibited the protein expressions compared with model group. Conclusion: Traditional Chinese medicine glabridin significantly ameliorated the lung injury induced by LPS in rats via reducing inflammation which caused by the inhibition of p38MAPK and ERK signaling pathway and antioxidant effect.

Tempol, a membrane-permeable radical scavenger, ameliorates lipopolysaccharide-induced acute lung injury in mice: A key role for superoxide anion

bjective To investigate the role of extracellular signal-regulated kinases（ERK） signal pathway in the lipopolysaccharide（LPS）-induced acute lung injury（ALI） in rats. Methods One hundred and twenty male SD rats were randomly divided into four groups（n=30）： Saline group, LPS group, LSP＋U group and Saline＋U group. Each group was divided into five subgroups at 1, 3, 6, 12 h and 24 h time point respectively. Western blot was used to detect the expression of phosphorylation of ERK1/2 and high-mobility group box 1（HMGB-1） in lung tissues, nuclear factor-κB（NF-κB）p65 in the nuclear extracts, which reflected the extent of lung injury. Additionally we examined the concentration of tumor necrosis factor（TNF）-α, interleukin（IL）-10 and inducible nitric oxide synthase（iNOS） in bronchoalveolar lavage fluid（BALF）, the lung water content and the histopathologic changes of lung. In addition, survival rate was investigated between LPS group and LSP＋U group,and each group with other 10 rats. Results With the administration of LPS, the phospho？蛳ERK1/2 substantially increased immediately and subsequently the expression of NF？蛳κB in the nuclear extracts was increased significantly and reached its peak level at 3 h and 12 h. Moreover, HMGB-1, one of the key mediators in the development of sepsis increased significantly after LPS administration. The concentrations of TNF？蛳α,iNOS and IL-10 were also increased. Pathological examination showed that the normal structure of lung was destroyed badly after LPS injection. U0126 effectively inhibited the activation of ERK1/2, blocked LPS-induced NF-κB activation and HMGB-1 expression in lung tissue, reduced the lung water content[（4.59±0.51） vs （5.19±0.10）, P〈0.05], the concentration of TNF-α, iNOS and IL-10[TNF-α, reached a peak at 3 h,（28±3） vs （70±10）（P〈0.05）. iNOS, reached a peak at 12 h, （7 771±957） vs （10 679±1 641）,P〈0.05. IL-10,reached a peak at 6 h,（59±10） vs（91±11）, P〈0.05], and prevented LPS-induced lung injury. Conclusions ERK1/2 plays an important role in the development of LPS-induced ALI, and the activation of ERK1/2 may be one of the mechanisms of LPS-induced ALI. Key words: Extracellular signal-regulated kinases; Nuclear factor-κB; Acute lung injury; Tumor necrosis factor-α; Interleukin-10; Inducible nitric oxide synthase; Lipopolysaccharide

To investigate the key role of nuclear factor E2-related factor 2 (Nrf2) in the treatment of lung injury in sepsis mice by regulating Nrf2/heme oxygenase-1 (HO-1)/high mobility group protein B1 (HMGB1) pathway. 120 male wild type (WT) and 120 Nrf2 knockout (Nrf2-KO) ICR mice were randomly divided into Sham group, H2 control group (Sham+H2 group), cecal ligation and puncture (CLP) induced sepsis model group (CLP group) and H2 intervention group (CLP+H2 group), with 30 mice in each group. The sepsis model was reproduced by CLP. The same operation was done in Sham group and Sham+H2 group except CLP. The mice in Sham+H2 group and CLP+H2 group were challenged by 2% H2 for 1 hour at 1 hour and 6 hours after operation respectively, while the mice in Sham group and CLP group only inhaled air. Twenty mice in each group were collected to observe the 7-day survival. The other mice were sacrificed at 24 hours after the reproduction of model, and the lung tissues were harvested. The activities of superoxide dismutase (SOD) and catalase (CAT) and malondialdehyde (MDA) contents were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of HO-1 and HMGB1 were determined by Western Blot, and the positive expression of HO-1 was also detected by immunofluorescence. Compared with Sham groups, the 7-day survival rates of WT and Nrf2-KO mice in CLP groups were significantly lowered [WT: 0% (0/20) vs. 100% (20/20), Nrf2-KO: 0% (0/20) vs. 100% (0/20), both P < 0.05]; the 7-day survival rates of CLP+H2 group in WT mice were significantly higher than those of CLP group [40% (8/20) vs. 0% (0/20), P < 0.05], but there was no significant difference between CLP+H2 group and CLP group in Nrf2-KO mice [0% (0/20) vs. 0% (0/200), P > 0.05]. In WT mice, compared with Sham group, the activities of SOD and CAT in lung tissue of CLP group were decreased significantly [SOD (kU/g): 131.30±28.21 vs. 251.00±22.84, CAT (kU/g): 13.43±1.52 vs. 20.76±1.63, both P < 0.01], the MDA content, the expressions of HO-1 and HMGB1 were increased significantly [MDA (μmol/g): 6.26±1.18 vs. 4.16±0.58, HO-1/β-actin: 0.160±0.045 vs. 0.023±0.005, HMGB1/β-actin: 0.656±0.055 vs. 0.005±0.001, all P < 0.05]. Compared with CLP group, the activities of SOD, CAT and HO-1 expression in lung tissue of CLP+H2 group were significantly increased [SOD (kU/g): 220.32±35.06 vs. 131.30±28.21, CAT (kU/g): 18.95±2.49 vs. 13.43±1.52, HO-1/β-actin: 0.376±0.025 vs. 0.160±0.045, all P < 0.01], while the MDA contents and HMGB1 expressions were significantly decreased [MDA (μmol/g): 4.26±0.75 vs. 6.26±1.18, HMGB1/β-actin: 0.343±0.040 vs. 0.656±0.055, both P < 0.05]. In Nrf2-KO mice, compared with Sham group, the activity of CAT in CLP group was significantly lowered (kU/g: 12.28±1.49 vs. 19.11±1.53, P < 0.01), MDA contents and the expressions of HO-1 and HMGB1 were significantly increased [MDA (μmol/g): 6.85±0.54 vs. 4.59±0.50, HO-1/β-actin: 0.063±0.005 vs. 0.021±0.003, HMGB1/β-actin: 0.713±0.035 vs. 0.005±0.001, all P < 0.01], while there was no significant difference in SOD activity (kU/g: 114.19±9.94 vs. 135.75±28.10, P > 0.05). There was no significant difference in above parameters between CLP+H2 group and CLP group. H2 inhibits lung injury in septic mice through Nrf2/HO-1/HMGB1 pathway. Nrf2 plays a major role in the treatment of septic lung injury by H2.

Hydrogen gas reduces HMGB1 release in lung tissues of septic mice in an Nrf2/HO-1-dependent pathway

Propolis is a natural bee product, and it has many effects, including antioxidant, anti-inflammatory, antihepatotoxic, and anticancer activity. In this study, we aimed to explore the potential in vivo anti-inflammatory, antioxidant, and antiapoptotic properties of propolis extract on lipopolysaccharide (LPS)-induced inflammation in rats. Forty-two, 3- to 4-month-old male Sprague Dawley rats were used in six groups. LPS (1 mg/kg) was administered intraperitoneally to rats in inflammation, inflammation + propolis30, and inflammation+propolis90 groups. Thirty milligram/kilogram and 90 mg/kg of propolis were given orally 24 h after LPS injection. After the determination of the inflammation in lung and liver tissues by 18F-fluoro-deoxy-d-glucose-positron emission tomography (18FDG-PET), samples were collected. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), nitric oxide (NO), and DNA fragmentation were determined. The decrease of MDA levels in inflammation + propolis30 and inflammation + propolis90 groups was determined compared to the inflammation group in lung and liver tissues. The increase of SOD% inhibition in inflammation + propolis90 group was determined in liver, lung, and hemolysate compared to the inflammation group. Increased CAT activities in inflammation + propolis30 and inflammation + propolis90 groups were observed in liver tissue and hemolysate compared to inflammation group. In lung tissue, NO levels were lower in inflammation group compared to the control group, but DNA fragmentation levels were higher. 18F-FDG uptake of tissues in inflammation + propolis30 and inflammation + propolis90 groups was decreased compared to the inflammation group. In conclusion, the data of this study indicate that the propolis application may serve as a potential approach for treating inflammatory diseases through the effect of reducing inflammation and free oxygen radical production.

To investigate whether the G protein-coupled estrogen receptor (GPER) can attenuates acute lung injury in mice with exertional heat stroke (EHS) by inhibiting ferroptosis. Sixty SPF-grade male C57BL/6 mice were randomly divided into four groups: normal control group (control group), EHS model group (EHS group), dimethyl sulfoxide (DMSO) solvent group (EHS+DMSO group), and GPER-specific agonist G1 group (EHS+G1 group), with 15 mice in each group. All mice underwent 14 days of adaptive training at 24-26 centigrade before modeling, and the EHS model was established using a high-temperature treadmill device. After successful modeling, the mice were allowed to cool naturally at room temperature. In the EHS+G1 group, 40 μg/kg of the GPER-specific agonist G1 was slowly injected intraperitoneally immediately after modeling. In the EHS+DMSO group, 40 μg/kg of DMSO was slowly injected intraperitoneally immediately after modeling. The control group received no treatment. Five hours after modeling, abdominal aortic blood was collected, and lung tissues were harvested after euthanasia. The lung coefficient was calculated to evaluate lung injury. Lung histopathological changes were observed under a light microscope after hematoxylin-eosin (HE) staining, and a lung histopathological score was assigned. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), malondialdehyde (MDA), and Fe2+ in lung tissue. Immunofluorescence was used to detect the expression of glutathione peroxidase 4 (GPX4). Real-time polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of GPX4, ferroportin 1 (FPN1), and ferritin heavy chain 1 (FTH1). Western blotting was performed to detect the protein expression of GPX4, FPN1, and FTH1. Compared with the control group, the lung coefficient and lung histopathological score were significantly increased in the EHS group. HE staining showed significant thickening and unevenness of the alveolar septa and alveolar walls, partial alveolar collapse, and extensive erythrocyte, inflammatory cell, and plasma-like material extravasation in the alveolar spaces. Serum levels of TNF-α, IL-1β, MDA, and Fe2+ were significantly elevated. Immunofluorescence staining showed a significant decrease in GPX4-positive expression in lung tissue. Western blotting and RT-PCR showed significantly reduced protein and mRNA expression of GPX4, FPN1, and FTH1 in lung tissue. Compared with the EHS group, the EHS+G1 group showed a significant reduction in lung coefficient and lung histopathological score [lung coefficient (mg/g): 3.9±0.1 vs. 4.6±0.3, lung histopathological score: 4.2±0.2 vs. 6.9±0.2, both P < 0.05]. HE staining revealed reduced severity of lung tissue fluid extravasation, inflammatory infiltration, decreased hemorrhage, and less severe alveolar structural damage. Serum levels of TNF-α, IL-1β, MDA, and Fe2+ were significantly reduced [TNF-α (ng/L): 44.3±0.2 vs. 64.6±0.3, IL-1β (ng/L): 69.3±0.4 vs. 97.8±0.2, MDA (nmol/L): 2.8±0.3 vs. 3.6±0.5, Fe2+ (nmol/L): 0.021±0.004 vs. 0.028±0.004, all P < 0.05]. Immunofluorescence staining showed a significant decrease in GPX4-positive expression in lung tissue (fluorescence intensity: 35.53±2.41 vs. 16.45±0.31, P < 0.05). RT-PCR and Western blotting showed significantly increased mRNA and protein expression of GPX4, FPN1, and FTH1 in lung tissue [mRNA expression: GPX4 mRNA (2-ΔΔCt): 0.44±0.05 vs. 0.09±0.01, FPN1 mRNA (2-ΔΔCt): 0.77±0.17 vs. 0.42±0.14, FTH1 mRNA (2-ΔΔCt): 0.75±0.04 vs. 0.58±0.01; protein expression: GPX4/β-actin: 0.96±0.11 vs. 0.24±0.04, FPN1/β-actin: 1.26±0.21 vs. 0.44±0.14, FTH1/β-actin: 0.27±0.12 vs. 0.15±0.07; all P < 0.05]. However, there were no statistically significant differences in any of the above indicators between the EHS+DMSO group and the EHS group. Activation of GPER can attenuate EHS-related lung injury in mice, and its mechanism may be related to the activation of the GPX4 signaling pathway and inhibition of ferroptosis.

This study was undertaken to evaluate the therapeutic potential effect of pentoxifylline (PTX) against arsenic trioxide (ATO)-induced cardiac oxidative damage in mice. Thirty-six male albino mice were divided into six groups and treated intraperitoneally with normal saline (group 1), ATO (5 mg/kg; group 2), PTX (100 mg/kg; group 3), and different doses of PTX (25, 50, and 100 mg/kg; groups 4, 5, and 6, respectively) with ATO. After four weeks, the blood sample was collected for biochemical experiments. In addition, cardiac tissue was removed for assessment of oxidative stress markers and histopathological changes (such as hemorrhage, necrosis, infiltration of inflammatory cells, and myocardial degeneration). The findings showed that ATO caused a significant raise in serum biochemical markers such as lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and troponin-I (cTnI), glucose, total cholesterol (TC), and triglyceride (TG) levels. In addition to histopathological changes in cardiac tissue, ATO led to the significant increase in cardiac lipid peroxidation (LPO) and nitric oxide (NO); remarkable decrease in the activity of cardiac antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx); and the depletion of the total antioxidant capacity (TAC) and total thiol groups (TTGs). PTX was able to reduce the increased levels of serum cardiac markers (LDH, CPK, cTnI, TC, and TG), cardiac LPO, and improve antioxidant markers (TAC, TTGs, CAT, SOD, and GPx) alongside histopathologic changes. However, no significant changes were observed in elevated serum glucose and cardiac NO levels. In conclusion, the current study showed the potential therapeutic effect of PTX in the prevention of ATO-induced cardiotoxicity via reversing the oxidative stress.

The aim of the present work was to investigate possible protective effects of febuxostat, a highly potent xanthine oxidase inhibitor, against acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats. Male Sprague Dawley rats were randomly divided into six groups, as follows: (i) vehicle control group; (ii) and (iii) febuxostat 10 and febuxostat 15 groups, drug-treated controls; (iv) LPS group, receiving an intraperitoneal injection of LPS (7.5 mg/kg); (v) and (vi) febuxostat 10-LPS and febuxostat 15-LPS groups, receiving oral treatment of febuxostat (10 and 15 mg/kg/day, respectively) for 7 days before LPS. After 18 h administration of LPS, blood was collected for C-reactive protein (CRP) measurement. Bronchoalveolar lavage fluid (BALF) was examined for leukocyte infiltration, lactate dehydrogenase (LDH) activity, protein content, and total nitrate/nitrite. Lung weight gain was determined, and lung tissue homogenate was prepared and evaluated for oxidative stress. Tumor necrosis factor-α (TNF-α) was assessed in BALF and lung homogenate. Moreover, histological changes of lung tissues were evaluated. LPS elicited lung injury characterized by increased lung water content (by 1.2 fold), leukocyte infiltration (by 13 fold), inflammation and oxidative stress (indicated by increased malondialdehyde (MDA), by 3.4 fold), and reduced superoxide dismutase (SOD) activity (by 34 %). Febuxostat dose-dependently decreased LPS-induced lung edema and elevations in BALF protein content, infiltration of leukocytes, and LDH activity. Moreover, the elevated levels of TNF-α in BALF and lung tissue of LPS-treated rats were attenuated by febuxostat pretreatment. Febuxostat also displayed a potent antioxidant activity by decreasing lung tissue levels of MDA and enhancing SOD activity. Histological analysis of lung tissue further demonstrated that febuxostat dose-dependently reversed LPS-induced histopathological changes. These findings demonstrate a significant dose-dependent protection by febuxostat against LPS-induced lung inflammation in rats.

The study was aimed to evaluate the effects of hydro-ethanol extract Zataria multiflora on the brain tissue oxidative damage, and hippocampal interleukin-6 (IL-6) as well as learning and memory capacity in lipopolysaccharide (LPS) - challenged rats. The rats were randomized into five groups as follow: Control group: Rats were treated with saline, LPS group: Rats were treated with LPS 1.00 mg kg-1, ZM50, ZM100 and ZM200 groups in which the rats were treated with Z. multiflora extract (50.00, 100 or 200 mg kg-1 per day, respectively). The treatments including extract or vehicle were administered intraperitoneally ‎and given three days before the behavioral tests and were continued within a6-day behavioral experiment. Injection of LPS was daily done before the behavioral tests. Finally, the brains were collected for biochemical evaluations. Although LPS administration prolonged the latency in Morris water maze and shortened the latency to enter the dark chamber in passive avoidance test, ZM extract restored these changes to approach control group values. Also, LPS increased IL-6, malondialdehyde (MDA) and nitric oxide (NO) metabolites levels and lowered thiol, superoxide dismutase (SOD) and catalase (CAT) levels in the brain, however, Z. multiflora extract reduced IL-6, MDA and NO metabolites concentrations, but increased thiol content, SOD, and CAT levels. The results of this study showed that Z. multiflora ameliorated learning and memory dysfunction in LPS - challenged rats by alleviating of inflammatory responses and brain tissue oxidative damage.