The mechanisms responsible for ischemia/reperfusion (I/R) injury have direct or indirect relevance to clinical lung injury after severe shock, cardiopulmonary bypass, and transplantation. This study investigated the effects of aspirin on intestinal I/R-induced acute lung injury (ALI) in rats. Lipopolysaccharide (LPS) induced cyclooxygenase-2 (COX-2) expression in A549 and RAW264.7 cells. RAW264.7 macrophages had shown greater expression of COX-2 than A549 cells. In addition, the NADPH oxidase inhibitor apocynin and p38 MAPK inhibitor SB203580 attenuated LPS-stimulated COX-2 expression. To induce ALI, intestinal ischemia was performed for 60 min prior to the 4 hr reperfusion by clamping the superior mesenteric artery in Sprague-Dawley rats. In order to test and compare the effect of non-specific COX inhibitor aspirin with the effect of mepacrine, a well known phospholipase$A_{2}$ inhibitor, rats were divided into 4 groups: Sham, I/R, Mepa+I/R (mepacrine, 60 mg/kg, i.p.), ASA+I/R (aspirin, 10 mg/kg, i.p.). In the present investigation, myeloperoxidase activities in the lung and intestinal tissues were increased by I/R. These changes were reduced by single pretreatment of mepacrine (60 mg/kg, i.p.) or aspirin (10 mg/kg, i.p.) 30 min before I/R. Structural studies demonstrated that the tissue injuries in the lung and intestine after I/R were also attenuated by the pretreatment of mepacrine or aspirin. These results suggest that I/R-induced ALI is mediated, in part, by the activation of COX. In addition, pretreatment of aspirin might be helpful for the prevention of ALI in ARDS-prone patients. In addition, the p38 MAPK inhibitor and apocynin also might be helpful to ALI through the inhibition of COX-2 expression.
Nikfarjam, Bahareh Abd;Adineh, Mohtaram;Hajiali, Farid;Nassiri-Asl, Marjan
Journal of Pharmacopuncture
/
v.20
no.1
/
pp.52-56
/
2017
Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.
Kim, Sung-Sam;Oh, Dae-Han;Choi, Se-Min;Kim, Kang-Woong;Kim, Kyoung-Duck;Lee, Bong-Joo;Han, Hyon-Sob;Lee, Kyeong-Jun
Korean Journal of Fisheries and Aquatic Sciences
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v.48
no.3
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pp.308-313
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2015
A $3{\times}3$ factorial study was conducted to investigate the effects of dietary protein and lipid levels on the growth, feed utilization and innate immunity of red seabream Pagrus major. Nine diets consisting of three protein levels (42%, 46% and 50% crude protein) and three lipid levels (10%, 14% and 18% crude lipid) were formulated. Triplicate groups of red seabream were fed the experimental diets to apparent satiation (5-6 times a day, from 08:00 to 18:00 h at 2-h intervals) for 10 weeks. At the end of the feeding trial, the weight gain and specific growth rate of fish fed P46L14 (46% protein and 14% lipid), P50L10 (50% protein and 10% lipid) and P50L14 (50% protein and 14% lipid) were significantly (P<0.05) higher than those of fish fed P42L18 (42% protein and 18% lipid). The feed conversion ratios (FCR) of the fish were affected by dietary lipid levels (P<0.039), but not dietary protein levels. The FCR tended to increase with increasing dietary lipid levels from 10% to 18% with the 46% and 50% protein levels. The weight gain, protein efficiency ratio, specific growth rate, feed intake and survival of fish were not affected by either dietary protein or lipid levels. Myeloperoxidase activity in the group fed P50L14 (50% protein and 14% lipid) was significantly higher than that in the group fed P42L10 (42% protein and 10% lipid) or P50L18 (50% protein and 18% lipid). However, the myeloperoxidase activity of fish was not affected by either dietary protein or lipid level. The fish fed P46L14 (46% protein and 14% lipid) and P46L18 (46% protein and 18% lipid) showed significantly higher superoxide dismutase activity than did the fish fed P46L10 (46% protein and 10% lipid), P50L10 (50% protein and 10% lipid) of P50L18 (50% protein and 18% lipid). In conclusion, the optimum protein and lipid levels for the growth and feed utilization of juvenile red seabream were 46% and 14%, respectively, and the optimum dietary protein to energy ratio was 27.4 g/MJ.
The efficacy of DA-6034, a new flavonoid derivative, was investigated in comparison with sulfasalazine in a trinitrobenzene sulfonic acid (TNBS)-induced rat colitis. Under light anaesthesia with ether, rats were subjected to intracolonic administration of 30mg TNBS in 50% ethanol (0.5ml) and were then sacrificed at 7 or 21 days after colitis induction. The TNBS control group (the saline treated colitic rat) exhibited ulceration and inflammation of the distal colon with formation of granuloma and pathologic connections. Moreover, an increase in colonic myeloperoxidase (MPO) activity (investigated as an index of leukocyte adhesion and accumulation) and an elevated colonic leukotriene $B_4$ ($LTB_4$) level were observed. The colitic rats received DA6034 (0.3-30mg/kg) or sulfasalazine (50-100mg/kg), prednisolone (0.3-3mg/kg) after the induction of colitis until they were sacrificed. Oral treatment with DA-6034 resulted in significant reductions of macroscopic colonic damage, colonic inflammation. DA6034 had a more potent effect than sulfasalazine and prednisolone on macroscopic colonic damage, while it has similar effect with prednisolone on the reduction of colonic $LTB_4$ synthesis and MPO activity. This study show, therefore, that DA-6034 is effective m attenuating the colonic lesion in an TNBS-induced colitis model. Furthermore, the results suggest that the effect of DA-6034 is partially related to its action on $LTB_4$ synthesis and MPO inhibition.
Joo, Minjae;Kim, Han Sang;Kwon, Tae Hoon;Palikhe, Alisha;Zaw, Tin Sandar;Jeong, Ji Hoon;Sohn, Uy Dong
The Korean Journal of Physiology and Pharmacology
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v.19
no.1
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pp.43-50
/
2015
It has been shown that the extracts including eupatilin and quercetin-3-${\beta}$-D-glucuronopyranoside had mucoprotective effects on the esophagus and stomach through their antioxidant activities. This study was designed to investigate the anti-inflammatory effect of these flavonoid compounds in an animal model of inflammatory bowel disease induced by 2,4,6-trinitrobenzene sulfonic acid. Experimental colitis was induced by intracolonic administration of 2,4,6-trinitrobenzene sulfonic acid. Extracts including eupatilin or quercetin-3-${\beta}$-D-glucuronopyranoside were orally administered to animals 48, 24, and 1 h prior to the induction of colitis and then again 24 h later. The animals were sacrificed 48 h after by 2,4,6-trinitrobenzene sulfonic acid treatment and the macroscopic appearance of the colonic lesions was scored in a blinded manner on a scale of 1 to 10. The inflammatory response to colitis induction was assessed by measuring myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, total glutathione levels, and malondialdehyde concentrations in the colon. The results indicated that extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside dose-dependently improved the morphology of the lesions induced by 2,4,6-trinitrobenzene sulfonic acid and reduced the ulcer index accordingly. In addition, rats receiving extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside showed significantly decreased levels of mucosal myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, and malondialdehyde levels, and increased total glutathione levels. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside ameliorated the inflammatory response and colonic injury in acute colitis by decreasing oxidative stress and neutrophil activation. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside may inhibit acute colitis.
Effects of wheat arabinoxylan on mouse spleen lymphocytes and peritoneal macrophages were examined in vitro. Among three wheat arabinoxylans (A1: low MW, A2: medium MW, A3: high MW), A3$(50{\sim}100\;{\mu}g/mL)$ increased the viability of spleen lymphocytes up to $114{\sim}125%$ of the control. A1 and A3 $(20\;{\mu}g/mL)$ increased the viability of lipopolysaccharide-treated lymphocytes synergistically. Viability of murine peritoneal macrophages treated with wheat arabinoxylans $(10{\sim}100{\mu}g/mL)$ was increased up to $135{\sim}175%$ of the control. The cytotoxic activity of macrophages against murine lymphocytic leukemic cell increased in the presence of wheat arabinoxylan. Phagocytic index of macrophages treated with wheat arabinozylans $(20\;{\mu}g/mL)$ significantly increased $197{\sim}232%$ compared with the control, and lysosomal phosphatase and myeloperoxidase activities also increased significantly (p<0.05). Treatment of wheat arabinoxylans tended to decrease nitrite production, but significantly stimulated $H_2O_2\;and\;O_2$ productions of macrophages (p<0.05). These results indicate that the immunostimulating effect of wheat arabinoxylan may be closely related with lysosomal enzyme activity and reactive oxygen intermediate production of macrophages.
Yangkyuksanhwa-Tang (YKSH), consisting of nine different herbs, is commonly used in Soyangin-type individuals with stroke, based on the Sasang Constitution Theory in Korea. However, no evidence has yet confirmed a beneficial effect of YKSH in ischemic stroke treatment. In this study, we investigated the effects of YKSH on ischemic brain injury in a mouse model of cerebral ischemia. Focal cerebral ischemia in mice was induced by photothrombosis, and behavioral recovery was evaluated. Infarct volume, inflammation, and newly generated cells were evaluated by histology and immunochemistry. YKSH treatment resulted in a significant recovery from the motor impairments induced by focal cerebral ischemia, as determined with wire grip and rotarod tests. YKSH treatment also decreased the infarct volume and the number of cells positive for tumor necrosis factor-${\alpha}$ and myeloperoxidase when compared with a vehicle-treated control group. By contrast, YKSH treatment considerably increased the number of cells positive for glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1, as well as the number of cells doubly positive for Ki67/doublecortin when compared with the vehicle-treated group. These results suggest that YKSH treatment attenuated the infarct size by anti-inflammatory action, astrocyte and microglia activation, and neuronal proliferation, thereby facilitating neurofunctional recovery from a cerebral ischemic assault. YKSH could therefore be a potential treatment for neurofunctional restoration of the injured brains of patients with stroke.
Objectives : This study was designed to evaluate the effects of Tokoro Rhizoma pharmat $KI_{10}$ in nephritis induced by lipopolysaccharide(LPS) in rat. Methods : Rats were divided into 4 groups and 3 groups(LPS, saline, Tokoro Rhizoma pharmacopuncture(TR-P) group) were injected LPS to induce nephritis. TR-P group was treated with TR at $KI_{10}$ three times for a week, saline group with normal saline. To evaluate the effects of TR at $KI_{10}$ on nephritis in rats, white blood cell(WBC), neutrophil in blood, creatinine, cytokine-induced neutrophil chemoattractant-1(CINC-1) in serum and urinary volume, creatinine in urine, renal myeloperoxidase(MPO) were measured and renal tissue was analyzed. Results : TR-P group significantly reduced WBC, neutrophil in blood, creatinine, CINC-1 in serum, creatinine in urine and renal MPO than LPS group. TR-P group increased urinary volume but, not significant. Conclusion : TR at $KI_{10}$ has a therapeutic effect on nephritis in LPS stimulated rat. Therefore, it is suggested that TR at $KI_{10}$ may be an useful therapeutics for nephritis in clinical field after further researches.
A splanchic artery occlusion for 90 min followed by reperfusion of the mesenteric circulation resulted in a severe form of circulatory shock characterized by endothelial dysfunction, severe hypotension, marked intestinal tissue injury, and a high mortality rate. The effect of rutin, a flavonoid having antiprostanoid, anti-inflammatory, antithrombotic, antioxidant effect, were investigated in a model of splanchnic artery occlusion (SAO) shock in urethane anesthetized rats. Occlusion of the superior mesenteric artery for 90 min produced a severe shock state resulted in a fatal outcome within 120 min of reperfusion in many rats. Rutin was given as a bolus (1.28 mg/kg) 10 min prior to reperfusion. Administration of rutin significantly improved mean arterial blood pressure in comparison to vehicle treated rats (p<0.05). Rutin treatment also resulted in a significant attenuation in the increase in plasma amino nitrogen concentration, intestinal myeloperoxidase activity, intestinal lipid peroxidation, infiltration of neutrophils in intestine and thrombin induced adherence of neutrophils to superior mesentric artery segments. These results suggest that rutin provides beneficial effects in part by preserving endothelial function and attenuating neutrophil accumulation in the ischemic reperfused splanchnic circulation.
This study was conducted to investigate an effect of Mori Cortex in the cardiac injury following dermal scald burn in rats. Sprague-Dawley rats were induced scald bum (15% of total body surface area). Heart was removed at 5 h postburn and examined with biochemical assay, ultrastructural observations and stereological analysis. The activity of serum aspartate aminotransferase and creatinine was increased at 5 h postburn compared with them of control. Administration of heat extracts of Mori Cortex after scald burn inhibited the production of KC (neutrophil chemoattractant cytokine) and increased the activity of protein kinase C (PKC) in heart tissue. The activity of myeloperoxidase (MPO) in heart tissue was decreased both at 5 h postburn and in case of Mori Cortex administration after scald burn. Ultrastructurally, many contraction bands and separation of intercalated disk induced by scald burn were decreased by administration of heat extracts of Mori Cortex. In stereological analysis, administration of Mori Cortex after scald burn resulted the volume densities of myofibril and mitochondria were increased compared with them of burn control. These data suggest that Mori Cortex may be a useful stuff to the range of available treatments for cardiac injury induced by skin burn.
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