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OBJECTIVE： To study the improvement and anti-inflammation mechanism of Shenfu injection on lung tissue of endotoxin shock model rats. METHODS： Totally 48 rats were randomized into control group，model group，dexamethasone group （positive control，1 mg/kg） and Shenfu injection low-dose，medium-dose and high-dose groups （5，10，15 mL/kg），with 8 rats in each group. Except for normal group， other groups were given intraperitoneal injection of lipopolysaccharide （LPS） to induce endotoxin shock model. After modeling， each group was given relevant medicine once intraperitoneally. 24 h after medication， HE staining was used to observe pathological changes of lung tissue in rats and pathological scoring was conducted. RT-PCR was used to determine mRNA levels of P65 and P50 proteins related to NF-κB signaling pathway. Western blot assay was used to determine the expression levels of P65 and P50 proteins in lung tissue， and the expression levels of P65 protein in nucleus and cytoplasm of lung tissue were also determined. The level of TNF-α in plasma in rats were determined by ELISA. RESULTS： Compared with control group， alveolar septum became thicker， obvious vascular engorgement was found， and a large number of neutrophils infiltrated the interstitium in model group. Histopathological score， mRNA and protein expression levels of P65 and P50 in lung tissues were increased significantly （P＜0.01 or P＜0.001）； the protein expression of levels P65 in nucleus and cytoplasm and level of TNF-α in plasma were increased significantly （P＜0.001）. Compared with model group， alveolar structure of rats in dexamethasone group and Shenfu injection medium-dose and high-dose groups was complete， no obvious bleeding was observed， and the degree of inflammatory cell infiltration was improved significantly. Histopathological score， mRNA and protein expression levels of P65 and P50 in lung tissue and level of TNF-α in plasma were decreased significantly （P＜0.05 or P＜0.01 or P＜0.001）. The protein expression level of P65 in nucleus and cytoplasm of lung tissue were decreased significantly in dexamethasone group and Shenfu injection low-dose and medium-dose groups were decreased significantly （P＜0.05 or P＜0.01 or P＜0.001）. CONCLUSIONS： Shenfu injection can decrease mRNA and protein expression levels of P65 and P50 in lung tissue， level of TNF-α in plasma， and protect lung tissue of endotoxin shock rats.
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Previously, we have reported that high mobility group box 1 (HMGB1), a proinflammatory mediator in sepsis, is released via the IFN-beta-mediated JAK/STAT pathway. However, detailed mechanisms are still unclear. In this study, we dissected upstream signaling pathways of HMGB1 release using various molecular biology methods. Here, we found that calcium/calmodulin-dependent protein kinase (CaM kinase, CaMK) is involved in HMGB1 release by regulating IFN-beta production. CaMK inhibitor, STO609, treatment inhibits LPS-induced IFN-beta production, which is correlated with the phosphorylation of interferon regulatory factor 3 (IRF3). Additionally, we show that CaMK-I plays a major role in IFN-beta production although other CaMK members also seem to contribute to this event. Furthermore, the CaMK inhibitor treatment reduced IFN-beta production in a murine endotoxemia. Our results suggest CaMKs contribute to HMGB1 release by enhancing IFN-beta production in sepsis.
Subject(s)
Benzimidazoles , Cytokines , Endotoxemia , HMGB1 Protein , Inflammation , Interferon Regulatory Factor-3 , Interferon-beta , Molecular Biology , Naphthalimides , Phosphorylation , Phosphotransferases , Protein Kinases , Sepsis , Signal TransductionABSTRACT
@#Objective To explore the effect of the dexamethasone on the levels of cytokine in the brain of endotoxin shock rats. Methods 54 Sprague-Dawley rats were randomized to the sham group, LPS group (lipopolysaccharide 8 mg/kg, i.v.) and DEX group (dexamethasone 5 mg/kg, i.v., in addition). The blood pressure was measured dynamically. The contents of interleukin (IL)-4, IL-10 and tumor necrosis factor (TNF)-α were detected with ELISA 1 h, 3 h, 6 h after shock. Results As the occurrence and development of shock, blood pressure went down gradually, IL-4 and IL-10 decreased (P<0.01), and TNF-α increased (P<0.01) in LPS group, while the TNF-α decreased (P<0.01), the IL-4 and IL-10 increased (P<0.01) in the DEX group compared with the LPS group. Conclusion Dexamethasone may protect the brain from the endotoxin shock injury in rats through regulating the inflammatory factor.
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Objective To test the role of a bioartificial kidney which consisted of a continuous vein-vein hemofiltration (CVVH) system and renal proximal tubule cells (PTCs)device, also called the renal tubule assist device (RAD),in endotoxin shock.Methods Female crossbred pigs 25 to 30 kg received with 3×1011 bacteria / kg body weight of E.coli intraperitoneally followed by treatment with a sham RAD without cells (n=7) or a RAD with cells (n=7). Data on blood pressure, cardiac output, renal blood flow and systemic marker of inflammation (IL-6) were measured.Results Cardiac outputs and renal blood flows were higher in the RAD group with cells than in the sham RAD group after E.coli infusion (P＜0.05). Plasma IL-6 levels were lower in the cell RAD group than in the sham RAD group after bacterial administration (P＜0.05). A significant difference on survival time was also observed between the two groups,with(9.07 ± 0.88) h in the cell RAD group vs. (5.10 ± 0.46)h in the sham RAD group, P＜ 0.01.Conclusion The bioartificial kidney containing PTCs can improve cardiovascular function,reduce cytokine levels,and prolong survival time.
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Objective To study the effect of FK506 pretreatment on endotoxin shock induced by LPS. Methods 155 mice of the Kunming strain were randomly divided into LPS group (n=80) and LPS|FK group (n=75). An endotoxin shock model of mice was reproduced by an intraperitoneal injection of LPS (25ml/kg for each mouse). Beginning from 2 days before the reproduction of the model, each mouse of the LPS+FK group were given 20ml/kg of FK506 q 12 h, by gastric gavage while animals of the LPS group were given an equal amount of 0.9% sodium chloride solution in the same way. The survival time of 20 mice of the LPS group and 15 mice of LPS+FK group was observed. 10 mice of each group were sacrificed at 0,0.5,2,4,6 and 8 hours after the injection of LPS, respectively. Blood samples were collected and serum TNF-? and IL-1?, contents, and activiry of ALT and AST were determined. The ratio of the wet weight and dry weight (W/D) of the lung was calculated, and the histopathology of the lung and the liver was studied. Results The mean survival time was longer and the 72h survival rate in mice of LPS+FK group was higher, but the serum ALT and AST activity and serum TNF-? and IL-1? contents were lower than those of LPS group at each time point. The W/D ratio of the lung in animals of both groups at 4,6 and 8 hours after the injection of LPS was greater than that at 0 hour (P
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To estimate the protective effect of removing circulatory TNF-a immunoadsorpnon or Amberlite XAD-7 absorption respectively on endotoxin shock. Methods New Zealand white rabbits injected with lethal dose of endotoxin underwent hemoper-fusion through immunoadsorbent or Amberh'te XAD-7 respectively. Plasma TNF-a levels were detected, and the sorbent efficiency and survival rate were observed. Results After 2 hours hemoperfusion through immunoadsorbent or Amberlite XAD-7 respectively, plasma TNF levels were significandy lower than those in die control group, especially, die sorbent efficiency of immunoadsorp-tion was higher dian dial of Amberlite XAD-7 adsorption. Conclusion Compared widi die nonspecific Amberlite XAD-7 adsorption, immunoadsorption might be a specific and more effective therapy of endotoxin shock.
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The effects of fructose 1, 6-diphosphate (FDP)on the hemodynamic variations in twelve dogs with endotoxin shock were observed. Following anesthesia,arterial and Swan-Gang catheters were inserted correspondingly into femoral and pulmonary arteries At 30mins and 90 mins following intravenous injection of endotoxin(IIE),these subjects were randomly assigned to receiving intravenous infusion of FDP at a dosage of 375mg/kg(group I,n= 6) or the equivalent volume of saline as control(group ,n= 6) The hemodynamic parameters of both groups were measured before IIE and every hour during 8 hours after IIE respectively. The results demonstrated that before IIE,there were not significant differences in the values of all parameters between both groups;im- mediately followillg IIE, the values of CI, MAP, PAMP, PCWP, SVI, LSWI, RSWI markedly decreased in both groups (P
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The plasma histamine concentration was elevated from the control level of 1.16? 0.28ng/ml to 11.42?5.2ng/ml 2-5 min after intravenous injection of Serratia mar-cescens endotoxin,and then declined gradually.The level was still slightly higher than the control 4 h later. The plasma noradrenaline(NA)and adrenaline (Ad) concentrations were increased from the control value of 1.13?0.28ng/ml and 1.05 ?0.32ng/ml to 7.33?1.86ng/ml and 10.04? 3.94ng/ml respectively. Six h later they were still significantly elevated.Arterial blood pressure was reduced to 24.16%of the control level 2-5 min after injection and to 69.2% 6 h later.Both cardiac output and cardiac index were decreased, but the peripheral resistance was increased during endotoxin shock. The significant dilatation of arterioles, venules and capillaries of bulbar conjunctiva microcirculation occurred 1-2 min after endotoxin injection, and the blood flow slowed down at the same time.Five min - six h after endotoxin injection arterioles constricted markedly, but capillaries and venules were still congested and distended.From this study, the interrelationship of some changes in the development of endotoxin shock were discussed.Endotoxin caused the congestion in microcirculatiolelen, reduction of blood returning to the heart and reduced peripheral resistance owing to the histamine released in large amounts. Drastic drop in blood pressure resulted in the increase of NA and Ad release reflexly, which led to the constriction of arterioles, the increase of the peripheral resistance and the restoration of the blood pressure to some extent.Since venules and capillaries were distended and congested,blood pressure and cardiac output still remained at lower level all the time during endotoxin shock.