Early Axonal Injury and Delayed Cytotoxic Cerebral Edema are Associated with Microglial Activation in a Mouse Model of Sepsis.

Sepsis-induced brain injury is associated with an acute deterioration of mental status resulting in cognitive impairment and acquisition of new functional limitations in sepsis survivors. However, the exact nature of brain injury in this setting is often subtle and remains to be fully characterized both in preclinical studies and at the bedside. Given the translation potential for the use of magnetic resonance imaging (MRI) to define sepsis-induced brain injury, we sought to determine and correlate the cellular changes with neuroradiographic presentations in a classic murine model of sepsis induced by cecal ligation and puncture (CLP). Sepsis was induced in 6-10-week-old male C57/BL6 mice by CLP. We used immunohistochemistry (IHC) to define neuropathology in a mouse model of sepsis along with parallel studies using MRI, focusing on cerebral edema, blood-brain barrier (BBB) disruption, and microglial activation on days 1 and 4 days after CLP. We demonstrate that septic mice had evidence of early axonal injury, inflammation, and robust microglial activation on day 1 followed by cytotoxic edema on day 4 in the cortex, thalamus, and hippocampus in the absence of BBB disruption. We note the superiority of the MRI to detect subtle brain injury and cytotoxic cerebral edema in comparison with the traditional gold standard assessment, i.e., percent brain water (wet-dry weight method). We conclude that inflammatory changes in the septic brain can be detected in real time, and further studies are needed to understand axonal injury and the impact of inhibition of microglial activation on the development of cerebral edema.

Early hyperbaric oxygen therapy improves survival in a model of severe sepsis.

Sepsis is a major clinical challenge, with therapy limited to supportive interventions. Therefore, the search for novel remedial approaches is of great importance. We addressed whether hyperbaric oxygen therapy (HBOT) could improve the outcome of sepsis using an acute experimental mouse model. Sepsis was induced in male CD-1 mice by cecal ligation and puncture (CLP) tailored to result in 80-90% mortality within 72 h of the insult. After CLP, mice were randomized into two groups receiving HBOT or not at different times after the initial insult or subjected to multiple HBOT treatments. HBOT conditions were 98% oxygen pressurized to 2.4 atmospheres for 1 h. HBOT within 1 h after CLP resulted in 52% survival in comparison with mice that did not receive the treatment (13% survival). Multiple HBOT at 1 and 6 h or 1, 6, and 21 h displayed an increase in survival of >50%, but they were not significantly different from a single treatment after 1 h of CLP. Treatments at 6 or 21 h after CLP, excluding the 1 h of treatment, did not show any protective effect. Early HBO treatment did not modify bacterial counts after CLP, but it was associated with decreased expression of TNF-α, IL-6, and IL-10 expression in the liver within 3 h after CLP. The decrease of cytokine expression was reproduced in cultured macrophages after exposure to HBOT. Early HBOT could be of benefit in the treatment of sepsis, and the protective mechanism may be related to a reduction in the systemic inflammatory response.

Tephrosin attenuates sepsis induced acute lung injury in rats by impeding expression of ICAM-1 and MIP-2.

Acute lung injury (ALI), a devastating form of respiratory infections, is characterized by increased edema, release of cytokines, weakened arterial oxygenation and infiltration of neutrophils and lymphocytes. The objective of the research envisaged was to reveal protective effects of tephrosin (TP) in ALI. In the present investigation, sepsis was triggered in rats by cecal ligation and puncture (CLP) method, and TP was administered intraperitonially. Five groups - Group A (control), Group B (Sham group) Group C (infected and untreated), and Group D and E (infected and treated with 25 and 50 mg/kg TP respectively) - of ten rats each, were used for the investigation. Evaluation parameters included measurement of arterial oxygenation, lung water content, protein determination, cytokine determination, neutrophil and lymphocyte count in the bronchoalveolar lavage fluid (BALF). As indicated by histopathological examination, the lung injury score was maximum in group C, but indicated reduction in group D and E. Intracellular adhesion molecule (ICAM)-1 and macrophage inflammatory protein-2 (MIP-2) are known to be important mediators responsible for ALI. Reduction in the ICAM-1 and MIP-2 expression was found to reduce after treatment with TP. In comparison to group D, group E reflected higher magnitude of ICAM-1 and MIP-2 suppression due to administration of higher TP dose. Compared to Group A and B, Group E indicated slightly higher expression of ICAM-1 and MIP-2. The research envisaged thus supports that TP attenuates ICAM-1 and MIP-2 expression in sepsis induced ALI rat model.

Development of Noninvasive in Vivo Approach to Assess Vascular Permeability in Inflammation Using Fluorescence Imaging.

INTRODUCTION: In vivo fluorescence imaging can quantify vascular permeability without requiring sacrifice of animals. However, use of this noninvasive approach for vascular permeability assessment in remote organ injury caused by systemic inflammatory disease has not been reported. METHODS: Evans blue (EB) and Genhance 750 fluorescent dye were mixed and injected into mice. The lung as a remote organ and the footpad as a noninvasive observational site were assessed in a cecal ligation and puncture (CLP)-induced systemic inflammation mouse model and compared with sham and hydrocortisone pretreated (CLP + HC) mouse models. Extraction of EB in harvested tissues was assessed as a conventional indicator of vascular permeability. Fluorescent intensities in the footpad or harvested lung were assessed and their correlation was analyzed to investigate this novel, noninvasive approach for estimation of lung vascular permeability. RESULTS: Fluorescent intensity in the footpad and harvested lung in the CLP group was significantly higher than in the other groups (footpad, sham vs. CLP, P < 0.0001; CLP vs. CLP + HC, P = 0.0004; sham vs. CLP + HC, P = 0.058; lung, sham vs. CLP, P < 0.0001; CLP vs. CLP + HC, P < 0.0001; sham vs. CLP + HC, P = 0.060). The fluorescent intensity in the footpad was strongly correlated with that in the lung (r = 0.95). CONCLUSIONS: This fluorescent technique may be useful for vascular permeability assessment based on EB quantification. Footpad fluorescent intensity was strongly correlated with that in the lung, and may be a suitable indicator in noninvasive estimation of lung vascular permeability.

Tussilagone Inhibits the Inflammatory Response and Improves Survival in CLP-Induced Septic Mice.

Tussilagone, extracted from Tussilago farfara is an oriental medicine used for asthma and bronchitis. We investigated its mechanism of action, its inhibitory effects on lipopolysaccharide-induced inflammation in macrophages, and its impact on viability in a cecal ligation and puncture (CLP)-induced mouse model of sepsis. Tussilagone suppressed the expression of the inflammatory mediators, nitric oxide and prostaglandin E2, and the inflammatory cytokines, tumor necrosis factor-alpha (TNF-α) and high-mobility group box 1 (HMGB1), in lipopolysaccharide-stimulated RAW 264.7 cells and peritoneal macrophages. Tussilagone also reduced the activation of the mitogen-activated protein kinases and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) involved in the activation of various inflammatory mediators in activated macrophages. Moreover, tussilagone administration (1 mg/kg and 10 mg/kg) produced decreased mortality and lung injury in CLP-activated septic mice. Augmented expression of cyclooxygenase (COX)-2 and TNF-α in pulmonary alveolar macrophages of septic mice were attenuated by tussilagone administration. Tussilagone also suppressed the induction of nitric oxide, prostaglandin E2, TNF-α and HMGB1 in the serum of the septic mice. Overall, tussilagone exhibited protective effects against inflammation and polymicrobial sepsis by suppressing inflammatory mediators possibly via the inhibition of NF-κB activation and the MAP kinase pathway. These results suggest the possible use of tussilagone for developing novel therapeutic modalities for sepsis and other inflammatory diseases.

The effect of intraperitoneal n-acetylcysteine on postoperative adhesions in rat models.

AIM: In this study, we researched the effect of local administration of N-acetylcysteine (NAC) on postoperative intraabdominal adhesion formation in the rat models. METHODS: 20 female Wistar Albino rats which were 5-7 months old are used for the study. The rats were divided into two equal groups. Group one was administered saline solution (n=10) while group two was administered NAC (n=10) after caecal abrasion. They were dissected on postoperative tenth day and were examined macroscopically and microscopically for the adhesion formation. Intraperitoneal adhesion formation was scored blinded with Evans model. The most adherent bowel section was excised for histopathologic examination. Mann Whitney U test were used for statistical analysis. RESULTS: In Group one, all rats have had adhesions. None of the rats in Group two had either severe inflammatory cell reaction or dense interstitial fibrosis. Macroscopic adhesion formation and microscopic inflammatory cell reaction and interstitial fibrosis formation after surgery were less at the group two (NAC applied) (p<0.05, p<0.05, p<0.05). CONCLUSION: We believe that the intraperitoneal single dose usage of NAC may be promising for decreasing the postoperative intraabdominal adhesions. KEY WORDS: N-acetylcysteine, Postoperative adhesion, Rat, Fibrosis.

Electroacupuncture pretreatment with different waveforms prevents brain injury in rats subjected to cecal ligation and puncture via inhibiting microglial activation, and attenuating inflammation, oxidative stress and apoptosis.

Sepsis is associated with high morbidity and mortality. This study was to investigate the protective effects of electroacupuncture (EA) pretreatment with different waveforms on septic brain injury in rats and its mechanism. Male Sprague-Dawley rats were pretreated by EA with different waveforms (continuous wave, dilatational wave, or intermittent wave) at Baihui (GV20) and Tsusanli (ST36) acupoints for 30min, and underwent cecal ligation and puncture (CLP) or sham operation. The results showed that EA pretreatment with different waveforms improved survival rate, attenuated encephaledema, brain injury, neuronal apoptosis and cognitive dysfunction, and preserved blood-brain barrier (BBB). EA pretreatment decreased the production of tumor necrosis factor(TNF)-α, interleukin(IL)-6, malondialdehyde (MDA), and increased the activity of superoxide dismutase (SOD) and catalase (CAT) in serum and hippocampus at 48h after sham or CLP operation. Additionally, EA pretreatment downregulated the expressions of toll-like receptor-4 (TLR-4), nuclear factor-kappa B (NF-κB) and ionized calcium binding adaptor molecule 1(Iba 1). The effect of dilatational wave was the most significant, followed by intermittent wave, and continuous wave was relatively poor. In conclusion, our results demonstrate that EA pretreatment with three waveforms alleviates sepsis-induced brain injury by inhibition of microglial activation and attenuation of inflammation, oxidative stress and apoptosis. These findings suggest that EA pretreatment with dilatational wave at Baihui and Tsusanli acupoints might be a promising therapeutic strategy for relieving septic brain injury.

Cationic liposomes containing antioxidants reduces pulmonary injury in experimental model of sepsis: Liposomes antioxidants reduces pulmonary damage.

The intracellular redox state of alveolar cells is a determining factor for tolerance to oxidative and pro-inflammatory stresses. This study investigated the effects of intratracheal co-administration of antioxidants encapsulated in liposomes on the lungs of rats subjected to sepsis. For this, male rats subjected to sepsis induced by lipopolysaccharide from Escherichia coli or placebo operation were treated (intratracheally) with antibiotic, 0.9% saline and antioxidants encapsulated or non-encapsulated in liposomes. Experimental model of sepsis by cecal ligation and puncture (CLP) was performed in order to expose the cecum. The cecum was then gently squeezed to extrude a small amount of feces from the perforation site. As an index of oxidative damage, superoxide anions, lipid peroxidation, protein carbonyls, catalase activity, nitrates/nitrites, cell viability and mortality rate were measured. Infected animals treated with antibiotic plus antioxidants encapsulated in liposomes showed reduced levels of superoxide anion (54% or 7.650±1.263 nmol/min/mg protein), lipid peroxidation (33% or 0.117±0.041 nmol/mg protein), protein carbonyl (57% or 0.039 ± 0.022 nmol/mg protein) and mortality rate (3.3%), p value <0.001. This treatment also reduced the level of nitrite/nitrate and increased cell viability (90.7%) of alveolar macrophages. Taken togheter, theses results support that cationic liposomes containing antioxidants should be explored as coadjuvants in the treatment of pulmonary oxidative damage.

Effect of Carapa guianensis Aublet (Andiroba) and Orbignya phalerata (Babassu) in colonic healing in rats / Efeito da Carapa guianensis Aublet (Andiroba) eOrbignya phalerata (Babaçu) na cicatrização de colorrafias em ratos

Objective: to evaluate the healing effect of the babassu aqueous extract and andiroba oil on open wounds in the cecum of rats. Methods: fifty-four Wistar rats were divided into three groups of 18: 1) babassu group with application of aqueous extract of babassu; 2) andiroba group with application of the oil; and 3) control group, with application of saline solution. All procedures were done by gavage. Each group was divided into three subgroups of six animals according to the observation period of 7, 14 or 21 days. From each animal was removed caecum fragment of 1.5cm² diameter. The areas of the lesions were analyzed macroscopically and resected specimens by light microscopy using hematoxylin-eosin and Masson's trichrome. Results: abscess and infection were observed in two aroeira group animals, and in one only hematoma. In relationship to adhesions degree, babassu group had higher incidence of grade II while in the control and aroeira groups predominated adhesions grade I. On microscopic examination on day 7 fibroblast proliferation was greater in aroeira and lower in babassu group (p=0.028). On the 14th day polymorphonuclear were less pronounced in babassu (p=0.007). As for the resistance test of air insufflation, it was observed that in all andiroba group in all tested days showed be higher. As for collagen, on the 7th day it was present in 100% of animals of aroeira group. On the 14th day was more pronounced in the control group and at day 21 similar results were found in the control and aroeira groups. Conclusion: animals in babassu and andiroba groups showed better cecum healing compared to the control group.

Objetivo: avaliar o efeito cicatrizante do extrato aquoso do babaçu e do óleo de andiroba em feridas abertas no ceco de ratos. Métodos: cinquenta e quatro ratos Wistar foram divididos em três grupos de 18: 1) grupo babaçu, com aplicação do extrato aquoso de babaçu; 2) grupo andiroba, com aplicação do óleo; e 3) grupo controle, com aplicação de solução salina. Todos os procedimentos foram feitos por gavagem. Cada grupo foi dividido em três subgrupos de seis animais conforme o período de observação, aos 7, 14 ou 21 dias. De cada animal foi retirado fragmento do ceco com 1,5cm2 de diâmetro. As áreas das lesões foram analisadas por macroscopia e os segmentos ressecados das feridas por microscopia ótica em colorações de hematoxilina-eosina e tricrômico de Masson. Resultados: foram verificados abscesso e infecção em dois animais do grupo andiroba, e um com hematoma. Quanto ao grau de aderências, o grupo babaçu teve maior incidência de aderências grau II enquanto que no grupo controle e andiroba predominaram aderências grau I. Na análise microscópica no sétimo dia a proliferação fibroblástica foi maior no grupo andiroba e menor no grupo babaçu (p=0,028). No 14º dia os polimorfonucleares foram menos acentuados no grupo babaçu (p=0,007). Quanto ao teste de resistência à insuflação de ar atmosférico observou-se que o grupo andiroba em qualquer dos dias avaliados apresentou maior tensão. Quanto à colagenização, no sétimo dia, ela esteve presente em 100% dos animais do grupo andiroba. No 14º dia foi mais acentuada no grupo controle e no 21º dia resultados semelhantes para o grupo controle e andiroba. Conclusão: os animais dos grupos babaçu e andiroba apresentaram melhor cicatrização do ceco em comparação ao grupo controle.

Fish oil-supplemented parenteral nutrition could alleviate acute lung injury, modulate immunity, and reduce inflammation in rats with abdominal sepsis.

The objectives were to confirm that intravenous fish oil (FO) emulsions could alleviate acute lung injury, modulate immunity, and reduce inflammation in rats with abdominal sepsis and to explore the mechanisms of these effects. Thirty-six adult male Sprague-Dawley rats were divided into 4 groups randomly. Two days after central venous catheterization, rats were subjected to cecal ligation and puncture to produce abdominal sepsis. Rats were assigned to receive normal saline or total parenteral nutrition (TPN) containing standard soybean oil emulsions or FO-supplemented TPN at the onset of sepsis for 5 days. A sham operation and control treatment were performed in control group rats. Acute lung injury scores, peripheral blood lymphocyte subsets, plasma cytokines, and Foxp3 expression in the spleen were determined. Compared with the normal saline and TPN without FO, FO-supplemented TPN beneficially altered the distributions of the T-lymphocyte subsets and downregulated the acute lung injury scores, plasma cytokines, and expression of Foxp3 due to sepsis. Fish oil-supplemented TPN can decrease acute lung injury scores, alleviate histopathology, reduce the bacterial load in the peritoneal lavage fluid, modulate the lymphocyte subpopulation in the peripheral blood, downregulate Foxp3 expression in the spleen, and reduce plasma cytokines, which means that FO-supplemented TPN can alleviate acute lung injury, modulate immunity, and reduce inflammation in rats with abdominal sepsis.

Effect of Carapa guianensis Aublet (Andiroba) and Orbignya phalerata (Babassu) in colonic healing in rats.

OBJECTIVE: to evaluate the healing effect of the babassu aqueous extract and andiroba oil on open wounds in the cecum of rats. METHODS: fifty-four Wistar rats were divided into three groups of 18: 1) babassu group with application of aqueous extract of babassu; 2) andiroba group with application of the oil; and 3) control group, with application of saline solution. All procedures were done by gavage. Each group was divided into three subgroups of six animals according to the observation period of 7, 14 or 21 days. From each animal was removed caecum fragment of 1.5cm² diameter. The areas of the lesions were analyzed macroscopically and resected specimens by light microscopy using hematoxylin-eosin and Masson's trichrome. RESULTS: abscess and infection were observed in two aroeira group animals, and in one only hematoma. In relationship to adhesions degree, babassu group had higher incidence of grade II while in the control and aroeira groups predominated adhesions grade I. On microscopic examination on day 7 fibroblast proliferation was greater in aroeira and lower in babassu group (p=0.028). On the 14th day polymorphonuclear were less pronounced in babassu (p=0.007). As for the resistance test of air insufflation, it was observed that in all andiroba group in all tested days showed be higher. As for collagen, on the 7th day it was present in 100% of animals of aroeira group. On the 14th day was more pronounced in the control group and at day 21 similar results were found in the control and aroeira groups. CONCLUSION: animals in babassu and andiroba groups showed better cecum healing compared to the control group.

Dehydrocostuslactone inhibits LPS-induced inflammation by p38MAPK-dependent induction of hemeoxygenase-1 in vitro and improves survival of mice in CLP-induced sepsis in vivo.

We investigated the hypothesis that the administration of dehydrocostuslactone (DL), a sesquiterpene lactone found in Saussurea lappa Clarke (Compositae), might reduce organ failure and increase survival in a cecal ligation and puncture (CLP)-induced mouse model of sepsis due to HO-1 induction. Treatment of RAW264.7 cells with DL increased HO-1 expression in a time- and concentration-dependent manner, and this up-regulation of HO-1 by DL was significantly inhibited by silencing either Nrf2 and p38 or treating cells with SB203580 (a p38MAPK inhibitor), but it was not inhibited in the presence of SP600125 (an ERK inhibitor), PD98059 (a JNK inhibitor), or LY294002 (PI3K inhibitor). As expected, DL concentration dependently inhibited the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2), and the productions of NO and PGE2 in LPS-activated cells, and these inhibitions were reversed by silencing HO-1. Most importantly, administration of DL significantly reduced mortality and reduced serum IL-1ß and TNF-α and the infiltration of macrophages into liver tissues of CLP-mice. Inducible NOS expression in lung and liver tissues of CLP-mice was reduced by DL, which was reversed by the co-administration of zinc-protoporphyrin IX (ZnPPIX; a competitive inhibitor of HO-1). Our findings indicate that DL might be useful for the treatment of sepsis.

Treatment with the sphingosine-1-phosphate analogue FTY 720 reduces loss of plasma volume during experimental sepsis in the rat.

BACKGROUND: Increased vascular leakage leading to hypovolaemia and tissue oedema is common in severe sepsis. Hypovolaemia together with oedema formation may contribute to hypoxia and result in multiorgan failure and death. To improve treatment during sepsis, a potential therapeutic target may be to reduce the vascular leakage. Substances affecting the endothelial barrier are interesting in this respect, as it is suggested that increase in vascular leakage depends on reorganisation of the endothelial cells and breakdown of the endothelial barrier. The agonist of the bioactive lipid sphingosine-1-phosphate, FTY720, has been shown to modulate the integrity of the endothelium and reduce permeability both in vitro and in vivo. The aim of the present study was to determine if FTY720 could reduce the loss of plasma volume during experimental sepsis in rats. METHODS: Sepsis was induced by ligation and incision of the caecum in the rat. Plasma volume was determined before and 4.5 h after induction of sepsis by a dilution technique using (125) I-labelled albumin. RESULTS: FTY720 in a dose of 0.2 mg/kg reduced the loss of plasma during sepsis by approximately 30% compared with vehicle, without any adverse effects on haemodynamic and physiological parameters. The increase in hematocrit and haemoglobin concentration was also found to be higher in the vehicle group. CONCLUSION: FTY720 in a dose without haemodynamic side effects reduces loss of plasma volume during experimental sepsis most likely because of reduction in permeability and may therefore be beneficial in sepsis.

Ginsenoside Rg1 improves survival in a murine model of polymicrobial sepsis by suppressing the inflammatory response and apoptosis of lymphocytes.

BACKGROUND: Unbalanced inflammatory response and lymphocyte apoptosis are the main reasons for high mortality in patients with sepsis. Ginsenoside Rg1 (Rg1), the most important component isolated from Panax ginseng, has long been used to treat inflammatory and immune-related diseases. We designed this study to investigate the therapeutic effect of this agent on cecal ligation and puncture (CLP)-induced sepsis in mice. MATERIALS AND METHODS: We randomly divided C57BL/6 mice into four experimental groups: sham, sham plus Rg1, CLP, and CLP plus Rg1. We intravenously injected Rg1 (20 mg/kg) 1 h after CLP and evaluated survival, bacterial clearance, cytokine production, histology, neutrophil emigration, and lymphocyte apoptosis. RESULTS: Our study showed that treatment with Rg1 significantly improved survival in septic mice (P < 0.01). Rg1 administration suppressed the inflammatory response and enhanced bacterial clearance. Histologic examination of lung and liver showed only minor abnormalities in mice that received Rg1. In addition, Rg1 increased neutrophil counts in peritoneal cavity and inhibited lymphocyte apoptosis in thymus and spleen. CONCLUSIONS: Ginsenoside Rg1 has a protective role against CLP-induced polymicrobial sepsis by attenuating the proinflammatory response, enhancing innate immunity and preserving adaptive immunity. Rg1 could be a promising new agent for treatment of sepsis.

Ruptured uterus and bowel injury from manual removal of placenta: a case report.

BACKGROUND: Retained placenta is a significant cause of maternal mortality and morbidity throughout the developing world. 'Though, intestinal injury may arise as a complication of induced abortion following instrumentation through the genital tract, the involvement of the large bowel in complicated manual removal of placenta is a very rare occurrence CASE REPORT: We present the case of a 28 year-old Para 3+0, 3 alive woman who had attempted manual removal of placenta in a basic emergency obstetric care facility that resulted in lower uterine segment rupture with evisceration of bowels through the laceration outside the introitus. She subsequently had right hemi- colectomy with ileo-transverse anastomosis and repair of uterine rupture with bilateral tubal ligation. CONCLUSION: This case highlights the risk of exposing parturients to inexperienced attendants at delivery and emphasises the need for intensification of manpower training to attain the 5th MDG enunciated by the United Nations.

Effects of different pulmonary surfactants in the prevention of postoperative intraabdominal adhesion formation.

BACKGROUND: After abdominal surgery, the formation of postoperative adhesion is a serious problem. The aim of the study is to evaluate the efficacy of 2 different pulmonary surfactants, poractant and beractant, on adhesion prevention in an experimental model. MATERIALS AND METHODS: An experimental intraabdominal adhesion model was created in 18 adult female rats by cecal abrasion. The rats were randomly assigned to 3 groups. Group I received no further treatment, whereas groups II and III received intraperitoneal poractant and beractant, respectively, before closing the incision. On the 15th postoperative day, all rats underwent relaparotomy, intraabdominal adhesions were scored macroscopically according to Canbaz scoring system, and the cecum in each animal was evaluated microscopically. RESULTS: The median adhesion scores of group II and III rats were significantly lower when compared with group I (P = .02). Group III had a lower median adhesion score than did group II, but this did not reach significance (P > .05). CONCLUSION: These observations suggest that intraperitoneal instillation of both pulmonary surfactants is associated with lower adhesion scores, higher adhesion-free cases, and improved histologic findings.

Ethanol extract of Prunella vulgaris var. lilacina inhibits HMGB1 release by induction of heme oxygenase-1 in LPS-activated RAW 264.7 cells and CLP-induced septic mice.

The ethanol extract of the flower of P. vulgaris var. lilacina (EEPV) has been used traditionally as an antiinflammatory agent in many countries. Inducers of heme oxygenase-1 (HO-1) reduce high mobility group box 1 (HMGB1), a late phase cytokine, in sepsis. Although EEPV has been used as an antiinflammatory agent, no report is available as to whether it modifies HMGB1 in sepsis due to HO-1 induction. It was found that EEPV increased HO-1 protein expression in RAW 264.7 cells, which was significantly inhibited by LY294002, but not PD98059, SB203580 or SP600125. In addition, EEPV activated NF-E2-related factor (Nrf2) to move from the cytosol to the nucleus, and EEPV-induced HO-1 and activation of ARE-luciferase activity were significantly reduced by siNrf2 transfection and LY294002 but not SB203508. EEPV also significantly inhibited NF-κB luciferase activity, and decreased both iNOS/NO and COX-2/PGE(2) production in lipopolysaccharide (LPS)-stimulated macrophages which was reversed by siHO-1 RNA transfection. Importantly, EEPV inhibited HMGB1 release in LPS-activated macrophages in a PI3K-sensitive manner and reduced serum HMGB1 level and lung HMGB1 expression in cecal ligation and puncture (CLP)-induced septic mice. It is concluded that EEPV induces HO-1 expression through PI3K/Nrf2 signal pathways, which may be beneficial for the treatment of sepsis due to a reduction of HMGB1 release.

Saponin fraction from Astragalus membranaceus roots protects mice against polymicrobial sepsis induced by cecal ligation and puncture by inhibiting inflammation and upregulating protein C pathway.

Sepsis remains the leading cause of death in intensive care units. Uncontrolled systemic inflammation and an impaired protein C pathway are two important contributors to sepsis pathophysiology. Based on the beneficial effects of the saponin fraction from Astragalus membranaceus roots (SAM) against inflammation, liver dysfunction, and endothelium injury, we investigated the potential protective roles and underlying mechanisms of SAM on polymicrobial sepsis induced by cecal ligation and puncture (CLP) in mice. SAM, orally administered 1 h before and after CLP, significantly elevated the survival rate of mice. At 96 h after CLP operation, all mice in the model group died, whereas 33.3% of mice in the SAM (400 mg/kg)-treated group survived. SAM attenuated both inflammatory factors and their abilities to induce tissue dysfunction, which was mainly evidenced by decreased infiltration of polymorphonuclear leukocytes, tissue edema, and lung wet-to-dry weight ratio, lowered levels of myeloperoxidase (MPO), nitric oxide (NO), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in serum, as well as downregulated expressions of iNOS and IL-1beta mRNA in livers. Furthermore, we addressed the effects of SAM on the protein C (PC) pathway, closely linked with sepsis. In CLP-induced septic mice, SAM elevated the impaired expression of PC mRNA in livers. In vitro, SAM reversed the decreased expressions of thrombomodulin (TM) and endothelial PC receptor (EPCR) mRNA induced by lipopolysaccharide (LPS) in endothelial cells. These findings suggest that SAM is able to restore the impaired protein C pathway. Taken together, the current study demonstrates that SAM has protective effects on polymicrobial sepsis in mice. The mechanisms of action involve anti-inflammation and upregulation of the PC pathway.

Alpinia katsumadai Hayata prevents mouse sepsis induced by cecal ligation and puncture through promoting bacterial clearance and downregulating systemic inflammation.

Sepsis continues to be a challenge in clinic. Therapeutic strategies focus on local host defenses and the inhibition of overwhelming inflammation response. The present study aimed to investigate the protective effects and the underlying mechanisms of the ethanol extract of Alpinia katsumadai Hayata seeds (EAKH) on polymicrobial sepsis induced by cecal ligation and puncture (CLP) in mice. It was shown that oral administration of EAKH at 1 h before and 2 h after CLP significantly elevated the survival rate and the mean arterial pressure of mice. Histological examination and serum ALT/AST assessment demonstrated that EAKH protected the animals from lung and liver tissue injury and dysfunction. Although EAKH was devoid of direct bacteriostatic or bacteriocidal activities, it facilitated peritoneal bacteria clearance and increased leukocyte migration into peritoneal cavity of septic mice. Furthermore, EAKH remarkably decreased serum pro-inflammatory cytokine (TNF-alpha, IL-1beta and NO) levels in septic mice. These findings demonstrated that EAKH has preventive effects on mouse sepsis induced by CLP, which may be attributed to elevating local defense via promoting leukocyte migration to infection focus and attenuating systemic inflammation.

Sesame oil does not show accumulatively enhanced protection against oxidative stress-associated hepatic injury in septic rats.

BACKGROUND: Sepsis is one of the major causes of death reported in intensive care units. A daily supplement of sesame oil for 1 week significantly attenuates oxidative stress-associated hepatic injury in septic rats. However, the excess intake of sesame oil may be associated with a health risk. This study investigates the effect of accumulative sesame oil on oxidative stress-associated hepatic injury after cecal ligation and puncture in rats. METHODS: Sesame oil was administered daily (4 mL/kg/d, orally) to rats, and the total intake of sesame oil ranged from 0 (control) to 140 mL/kg before cecal ligation and puncture in 9 groups of rats. Oxidative stress was examined by determining the levels of lipid peroxidation and glutathione. Hepatic injury was evaluated by measuring serum levels of aspartate aminotransferase and alkaline phosphatase. RESULTS: Rats that received sesame oil for 4 and 5 weeks had a lower body weight gain compared with those that received saline. Lipid peroxidation was decreased in the 20-mL/kg and 28-mL/kg groups, but it was increased in the 140-mL/kg group compared with the control group. Glutathione levels were increased in the < or =28-mL/kg groups compared with the control group. Serum levels of aspartate aminotransferase and alkaline phosphatase were reduced in the < or =28-mL/kg groups compared with the control group. CONCLUSION: Sesame oil does not demonstrate accumulatively enhanced protection against oxidative stress-associated hepatic injury after cecal ligation and puncture in rats.