Effects of serum of the rats ventilated with high tidal volume on endothelial cell permeability and therapeutic effects of ulinastatin

Background With the widespread use of ventilators in treating critically ill patients, the morbidity of ventilator-induced lung injury (VILI) is increasing accordingly. VILI is characterized by a considerable increase in microvascular leakiness and activation of inflammatory processes. In this study we investigated the effects of inflammatory mediators in VILI rat serum on endothelial cytoskeleton and monolayer cellular permeability, as well as the therapeutic effect of ulinastatin, to explore the pathogenesis and the relationship between biotrauma and lung oedema induced by VILI. Methods Thirty healthy male Sprague-Dawley rats were randomly divided into three groups: group A (normal tidal volume ventilation), group B (high tidal volume ventilation) and group C (high tidal volume ventilation plus ulinastatin). The serum of each rat after ventilation was added to endothelial cell line ECV-304 medium for two hours to observe the effects of serum and/or ulinastatin on endothelial fibrous actin and permeability. Results Compared to rats ventilated with normal tidal volume, serum of rats ventilated with high tidal volume caused a striking reorganization of actin cytoskeleton with a weakening of fluorescent intensity at the peripheral filament bands and formation of the long and thick stress fibres in the centre resulting in endothelial contraction and higher permeability. Prior treatment with ulinastatin lessened the above changes significantly. The changes of permeability coefficient of endothelial permeability after group A, B or C rats serum stimulation were (6.95±1.66)%, (27.50±7.77)% and (17.71±4.66)% respectively with statistically significant differences (P<0.05) among the three groups. Conclusions The proinflammatory mediators in the serum of the rats given high tidal volume ventilation increases endothelial permeability by reorganizing actin cytoskeleton, and pretreatment with ulinastatin lessens the permeability by inhibiting of proinflammatory mediators.     

Actions of NO and INOS on Endotoxin Induced Rat Acute Lung Injury and Effect of Rhubarb on Them

This study is to explore the actions of nitric oxide(NO)and inducible nitric oxide synthase(iNOS)onendotoxin(lipopolysaccharide,LPS)induced rat acute lung injury(ALI)and effect of Rhubarb onthem.LPS was injected into the sublingual vein of male Wistar rats to prepare ALI animal models.Therats were divided into 4 groups:LPS,control,Rhubarb,and dexamethasone.Macroscopic andhistopathological examinations of the lung specimens were performed and the biological indexes oflung,including wet weight/dry weight,the rate of neutrophils and protein content in the pulmonaryalveolar lavage fluid,pulmonary vascular permeability and pulmonary alveolar permeability wereobserved.In the mean time,the contents of serum NO and the activities of lung tissue homogenateiNOS were measured.The results showed that in the LPS group,the injury and celluar infiltration inthe pulmonary stroma and alveoli were more prominent than that in the control group.Lung wetweight/dry weight,the rate of neutrophils,protein content,pulmonary alveolar permeability,pulmonary vascular permeability were significantly increased(P<0.01);NO and iNOS were alsomarkedly elevated(P<0.01).In the groups of dexamethasone and Rhubarb,the histopathologicalchanges were significantly milder,and all the above biological indexes of lung injury and the contentsof NO and the activities of iNOS were correspondingly decreased(P<0.05).The above datademonstrate that NO and iNOS play an important role in the onset of ALI;dexamethasone andRhubarb interfering treatment can ameliorate lung injury and decrease the concentrations of NO andthe activities of iNOS,showing that through inhibiting the levels of NO and the activities of iNOS,these2 agents exert protective effect on ALI induced LPS.     

Change of Th1/Th2 Cytokine Equilibrium in Rats with Severe Sepsis and Therapeutic Effect of Recombinant Interleukin-12 and Shenmai Injection(参麦注射液)

Objective: To evaluate the dynamic change of Th1/Th2 cytokines in serum, peritoneal lavage fluid (PLF) and splenic macrophages (SM) in rats with severe peritonitis, and to observe the therapeutic preparation. Methods: Severe peritonitis (SP) model was induced by intraperitoneal injection of E. coli and B. frag, and mild peritonitis (MP) model was induced by cecal ligation and punching. Then the following experiments were done: (1) Survival rates of animals after every 6 hrs in the 72 hrs after modeling were recorded, serum and PLF levels of cytokines, including tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ),and interleukin-10 (IL-10), 6 hrs, 12 hrs, 24 hrs and 48 hrs after modeling were measured. (2) Model rats were treated with rlL-12 or SMI, the survival rate was recorded and serum levels of TNF-α, INF-γ, and IL-10before and after treatment were measured, and (3) amount of these cytokines produced by SM were determined 6 hrs, 12 hrs and 24 hrs after treatment. The survival rates and levels of cytokines were then compared between the groups (model group treated with rlL-12 or SMI, untreated model group, and blank group). Results: Serum and PLF levels of IFN-γ, TNF-α at all the time points in SP rats were significantly lower than those in MP rats while those of IL-10 6 hrs and 12 hrs after modeling were significantly higher in the former than that in the latter ( P＜0.05). IFN-γ secretion of SM in SP rats was significantly higher than that in MP rats 6 hrs after modeling (P＜0.05). Administration of rlL-12 or SMI given before modeling could improve the survival rate of the model rats (P＜0.05) and cause significant increase of the serum level and SM secretion of IFN-γ. Conclusion: Imbalance in promoting/antagonizing inflammatory cytokines and Th2 response dominance appear in SP rats early at the initiating stage, and SM secretion of inflammation promoting factor also reduces. Administration in time of rlL-12 and SMI, may increase the survival rate, and its mechanism may be related with their immuno-stimulating action.     

Efficacy of Xuebijing Injection(血必净注射液)on Cardiopulmonary Bypass-Associated Pulmonary Injury:A Prospective,Single-center,Double Blinded,Randomized Controlled Trial

Objective: To evaluate the efficacy of Xuebijing Injection(血必净注射液, XBJ) on the lung injury induced by cardiopulmonary bypass(CPB). Methods: Fifty patients undergoing CPB were randomized to either the saline group or XBJ group according to a random number table(25 cases in each group). The patients in the saline group received saline and patients in XBJ group received XBJ at 12 h prior to the operation, at the beginning of the operation, and at 12 h after the second injection. The PaO_2/Fi O2 at extubation 3 days post-operation, duration of ventilation in the intensive care unit(ICU), and lengths of stay in the ICU and hospital were recorded. The levels of inflammatory mediators including interleukin(IL)-1β, IL-8, IL-10, and C-reactive protein(CRP) in bronchoalveolar lavage fluid(BALF) and plasma were measured. The neutrophil count and elastase neutrophil elastase in BALF were also measured. In addition, adverse events were monitored. Results: The PaO-2/FiO_2 in the XBJ group was higher than that in the saline group from 12 to 72 h post-operation(all P0.05). The blood levels of IL-1β, IL-8, and CRP in the XBJ group from 12 to 72 h were all significantly lower than those in the saline group(all P0.05). In contrast, the level of the anti-inflammatory cytokine IL-10 was significantly higher in the XBJ group than in the saline group(P0.05). In addition, 4 patients presented with atelectasis in the saline group and none in the XBJ group. Ten patients experienced mild acute respiratory distress syndrome(ARDS) during hospitalization, and 5 patients with mild ARDS were in the XBJ group(P0.05). Conclusion: XBJ shows protective potential against lung injury in patients who undergo CPB surgery, possibly through the downregulation of inflammatory mediators, reduction in neutrophil infiltration, and upregulation of IL-10(Trial registry: Chi CTR-TRC-14004628).     

Attenuation of Collagen Induced Arthritis by Centella asiatica Methanol Fraction via Modulation of Cytokines and Oxidative Stress

Objective To investigate the anti-inflammatory, antioxidant and anti-arthritic effects of Centella asiatica methanolfraction(CaM E) on collagen-induced arthritis(CIA), an animal model of rheumatoid arthritis. Methods Arthritis was induced in female wistar rats by immunization with porcine type II collagen. The CIA rats were treated orally with CaM E(50, 150, and 250 mg/kg/day) for 15 d(beginning on day 21 of the experimental period). The clinical, histological, biochemical, and immunological parameters were assessed. Results CaM E treatment(150 and 250 mg/kg) significantly attenuated the severity of CIA and reduced the synovial inflammation, cartilage erosion, and bone erosion as evident from both histological and radiographic data. The escalated plasma levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-12 alongwith nitric oxide in CIA rats decreased significantly on CaM E treatment. The serum levels of type-II collagen antibody were significantly lower in rats of CaM E(150 and 250 mg/kg) treated group than those in the arthritic group. Furthermore, by inhibiting the above mediators, CaM E also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage. Conclusion Our results clearly indicate that oral administration of CaM E suppresses joint inflammation, cytokine expression as well as antioxidant imbalance, thereby contributing to an amelioration of arthritis severity in CIA rats.     

Multiple Pharmacological Actions of Yiqi Huatan Decoction (益气化痰方) in A Model of Depression in Rats

Objective: To investigate the influence of Yiqi Huatan Decoction (益气化痰方, YHD) on a model of depression in rats under different pathological conditions. Methods: Thirty-two male SD rats were randomly divided into 4 groups of 8: normal, model, YHD, and maprotiline. The model group, YHD group and maprotiline group used separate feeding and rats were exposed to chronic and unpredictable stress to build the depression model. From day 2, the YHD group and maprotiline group were respectively given YHD (7 g/kg) and maprotiline (10 mg/kg) by gastrogavage once daily. The normal and model groups were given the same volume of drinking water. The medication duration were 21 days. At the end of the experiment, the serum levels of copper and zinc were determined by atomic absorption spectroscopy, plasma concentrations of adrenocorticotropic hormone (ACTH) and cortisol (COR) were detected by radioimmunoassay, and levels of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the hypothalamus were analysed by high performance liquid chromatography-eletricochemistry. Results: Compared with the content of copper and zinc in the serum of rats in the normal group, serum copper levels in model rats were significantly increased and zinc content was significantly reduced (both P<0.05). Plasma concentrations of ACTH and COR in the model group were significantly increased compared with those in the normal group (P<0.05, P<0.01). The contents of NE, DA, and 5-HT in the hypothalamus of rats in the model group were significantly reduced compared with those of the normal group (P<0.05 or P<0.01). Compared with those in the model group, the serum copper content and plasma concentrations of ACTH and COR were significantly decreased (all P<0.05); meanwhile, serum zinc content and hypothalamic contents of NE, DA, and 5-HT were significantly increased in rats of the YHD group (all P<0.05). The same effects were also shown in the maprotiline group except for 5-HT (all P<0.05). Conclusion: The pharmacological actions of YHD for depression might be related to improving trace-element anomalies, reversing endocrine dysfunction, and modulating the disorders of monoaminergic neurotransmitters.     

Efficacy of Xuebijing Injection (血必净注射液) on Cardiopulmonary Bypass-Associated Pulmonary Injury: A Prospective, Single-center, Double Blinded, Randomized Controlled Trial

Objective: To evaluate the efficacy of Xuebijing Injection (血必净注射液, XBJ) on the lung injury induced by cardiopulmonary bypass (CPB). Methods: Fifty patients undergoing CPB were randomized to either the saline group or XBJ group according to a random number table (25 cases in each group). The patients in the saline group received saline and patients in XBJ group received XBJ at 12 h prior to the operation, at the beginning of the operation, and at 12 h after the second injection. The PaO2/FiO2 at extubation 3 days post-operation, duration of ventilation in the intensive care unit (ICU), and lengths of stay in the ICU and hospital were recorded. The levels of inflammatory mediators including interleukin (IL)-1β, IL-8, IL-10, and C-reactive protein (CRP) in bronchoalveolar lavage fluid (BALF) and plasma were measured. The neutrophil count and elastase neutrophil elastase in BALF were also measured. In addition, adverse events were monitored. Results: The PaO2/FiO2 in the XBJ group was higher than that in the saline group from 12 to 72 h post-operation (all P<0.05). The blood levels of IL-1β, IL-8, and CRP in the XBJ group from 12 to 72 h were all significantly lower than those in the saline group (all P<0.05). In contrast, the level of the anti-inflammatory cytokine IL-10 was significantly higher in the XBJ group than in the saline group (P<0.05). In addition, 4 patients presented with atelectasis in the saline group and none in the XBJ group. Ten patients experienced mild acute respiratory distress syndrome (ARDS) during hospitalization, and 5 patients with mild ARDS were in the XBJ group (P<0.05). Conclusion: XBJ shows protective potential against lung injury in patients who undergo CPB surgery, possibly through the downregulation of inflammatory mediators, reduction in neutrophil infiltration, and upregulation of IL-10 (Trial registry: ChiCTR-TRC-14004628).     

Protective Effect of Rhubarb on Endotoxin-Induced Acute Lung Injury

To approac the mechanism of lipopolysaccharide(LPS)in causing acute lung injury(ALI)and theprotective effect of rhubarb and dexamethasone,lung specimens were examined with macroscopy,microscopy,electron microscopy and the biological markers of ALI including lung wet/dry weight,therate of neutrophils and protein content in the pulmonary alveolar lavage fluid,pulmonary capillarypermeability and pulmonary alveolar permeability index were observed.The mechanism of the ALI ismainly due to direct injury of alveolar epithelium and pulmonary vascular endothelium.Rhubarb anddexamethasone could significantly reduce the edema of the lung tissue,decrease the red blood cellexudation,neutrophil infiltration and plasma protein exudation in the alveoli and all the biologicalmarkers in comparison with the ALI model rats,indicating they have protective action on vascularendothelium and alveolar epithelium.     

Effect of ecoimmunonutrition supports on maintenance of integrity of intestinal mucosal barrier in severe acute pancreatitis in dogs

Background One of the major causes of death in severe acute pancreatitis （SAP） is severe infection owing to bacterial translocation. Some clinical studies suggested that ecoimmunonutrition （EIN） as a new strategy had better treatment effect on SAP patients. But the experiment studies on the precise mechanism of the effect of EIN were less reported. In this study, we mainly investigated the effects of EIN on bacterial translocation in SAP model of dogs. Methods SAP was induced by retrograde infusion of 5% sodium taurocholate into the pancreatic duct in healthy hybrid dogs. The SAP dogs were supported with either parenteral nutrition （PN） or elemental enteral nutrition （EEN） or EIN. The levels of serum amylase, serum aminotransferase and plasma endotoxin were detected before and after pancreatitis induction. On the 7th day after nutrition supports, peritoneal fluid, mesenteric lymph nodes （MLN）, liver, and pancreas were collected for bacterial culture with standard techniques to observe the incidence of bacterial translocation. Pathology changes of pancreas were analyzed by histopathologic grading and scoring of the severity of pancreas, and the degree of intestinal mucosal damage was assessed by measuring mucosal thickness, villus height, and crypt depth of ileum. Results Compared with PN and EEN, EIN significantly decreased the levels of serum amylase, serum aminotransferase, plasma endotoxin, and the incidence of bacterial translocation. Furthermore, compared with the others, the histology scores of inflammation in pancreas and the ileum injury （ileum mocosa thickness, villus height, and crypt depth） were significantly alleviated by EIN （P〈0.05）. Moreover, concerning liver function, the serum levels of alanine aminotransferase, aspartate aminotransferase and albumin were ameliorating significantly in the EIN group. Conclusion Our results suggested that EIN could maintain the integrity of intestinal mucosal barrier and reducing the incidence of bacterial translocation in SAP dogs. Early EIN was safe and more effective treatment for SAP dogs.     

Impaired upregulation of keratinocyte growth factor in injured lungs induced by Pseudomonas aeruginosa in immunosuppressed rats

Background The number of immunosupressed patients has increased in the past decades. Among them Pseudomonas aeruginosa （P. aeruginosa） is one of the leading bacteria for pneumonia that are associated with poor prognosis. However, the pathogenesis of P. aeruginosa pneumonia in immunosupressed patients is not understood completely. Previous reports showed keratinocyte growth factor （KGF） is associated with lung injury in immunocompetent hosts. In this study, we investigated the different reactions of lung injury, lung pathology and KGF expressions in P aeruginosa pneumonia between immunosuppressed and immunocompetent rats. Methods Immunosuppression of male rats was induced by injecting immunosuppressive subcutaneously. Pneumonia was established by instilling P aeruginous tracheally. The immunocompetent rats were the control group. Survival rate, lung histopathology, pulmonary permeability and oedema, KGF mRNA and protein expressions in lungs of both groups were investigated. Results The survival rate of immunosuppressed group was lower than that of immunocompetent group （33.3% vs 83.3%）. After exposure to bacteria, pulmonary permeability and wet/dry ratio in immunosuppressed group were higher than those in immunocompetent group. Pulmonary congestion and haemorrhage were more intensive in immunosuppressed group compared to immunocompetent group. Apoptosis and necrosis were also observed in infected lungs of immunosuppressed rats. Although we detected KGF expressions in lungs of both groups after infection, the expressions of KGF protein and mRNA gene in immunosuppressed group were much lower than in immunocompetent group. Conclusions Compared with immunocompetent group, there was more intensive lung injury in immunosuppressed group. Severe lung injury may contribute to the poor prognosis of pneumonia. KGF expressions of pneumonia in immunosuppressed rats were less than those in immunocompetent ones.     

Xuebijing Injection (血必净注射液) and Resolvin D1 Synergize Regulate Leukocyte Adhesion and Improve Survival Rate in Mice with Sepsis-Induced Lung Injury

Objective: To investigate the effect of combined application of Xuebijing Injection(血必净注射液, XBJ) and resolvin D1(RvD1) on survival rate and the underlying mechanisms in mice with sepsisinduced lung injury. Methods: The cecal ligation and puncture(CLP) method was used to develop a mouse sepsis model. Specific pathogen free male C57 BL/6 mice were randomly divided into 5 groups(n=20 each): sham, CLP, CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1. After surgery, mice in the CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1 groups were given XBJ(25 μL/g body weight), RvD1(10 ng/g body weight), and their combination(the same dose of XBJ and RvD1), respectively. In each group, 12 mice were used to observe 1-week survival rate, while the rest were executed at 12 h. Whole blood was collected for flow cytometric analysis of leukocyte adhesion molecules CD18, lung tissues were harvested for observing pathological changes, and testing the activity of myeloperoxidase(MPO) and the expression of intercellular cell adhesion molecule 1(ICAM-1). Results: Compared with the CLP group, the histopathological damage of the lung tissues was mitigated, MPO activity was decreased in the CLP+XBJ and CLP+RvD1 groups(P0.05). In addition, the 1-week survival rate was improved, proportion of CD18-expressing cells in whole blood and ICAM-1 protein expression in lung tissue were decreased in the CLP+XBJ+RvD1 group(P0.05 or P0.01). Conclusion: XBJ together with RvD1 could effectively inhibit leukocyte adhesion, reduce lung injury, and improve the survival rate of mice with sepsis.     

Effects of Acupuncture on Progesterone and Prolactin in Rats of Embryo Implantation Dysfunction

Objective:To investigate the effect of acupuncture on progesterone(P4) and prolactin(PRL) in rats of embryo implantation dysfunction(EID).Methods:On the first day of pregnancy,72 female Wistar rats were randomly allocated into the normal group,the EID model group,the acupuncture group and the P4 group(18 in each group).The normal group was injected sesame oil,while the other three groups were given mifepristone to establish the EID model.The acupuncture group and the P4 group were given treatment of acupuncture and P4 injection,respectively.The serum of P4 and PRL were detected by radioimmunoassay,and the mRNA and protein expressions of P4 receptor(PR) and PRL receptor(PRLR) were detected using real-time polymerase chain reaction and immunohistochemical method,respectively.Results:Compared with the normal group,the serum levels of P4 and PRL as well as the mRNA and protein expression levels of PR and PRLR in the EID model group were significantly lowered(P<0.01 or P<0.05).The above indices in the acupuncture group and the P4 group were significantly elevated compared with the EID model group(P<0.01 or P<0.05).Conclusion:Acupuncture can promote embryo implantation effectively,which might be related to the effects of acupuncture on upregulating the P4 and PRL levels in serum and the PR and PRLR expression levels in rats.     

Pulmonary artery perfusion with HTK solution prevents lung injury in infants after cardiopulmonary bypass

Background Pulmonary artery perfusion during cardiopulmonary bypass （CPB） is a novel adjunctive method, which can minimize the lung ischemic-reperfusion injury and inflammatory response. This study evaluated the protective effect of pulmonary perfusion with hypothermic HTK solution in corrections of congenital heart defects with pulmonary hypertension. Methods Between June 2009 and December 2009, 24 consecutive infants with congenital heart defects and pulmonary hypertension were randomly divided into perfused group （n=12） and control group （n=-12）. Oxygen index, alveolar-arterial 02 gradient, serum levels of malondialchehyche （MDA）, interleukin （IL）-6, -8, -10, soluble intercellular adhesion molecule-1 （slCAM-1）, and P-seiectin were measured before commencement and serially for 48 hours after termination of bypass. Results Oxygenation values were better preserved in the perfused group than in the control group. The serum levels of IL-6 increased immediately after CPB in both groups and returned to baseline at 48 hours after CPB, but it was restored faster and earlier in the perfused group. The serum levels of IL-8, slCAMol, and MDA remained at baseline at each point after CPB in the perfused group and elevated significantly immediately after CPB in the control group, except for slCAM-1 The serum level of IL-10 increased immediately after CPB and decreased to baseline at 48 hours after CPB in both groups, but the IL-10 level in the perfused group was significantly higher than in the control group at 12 hours after CPB. The serum P-selectin levels in the control group immediately after CPB were significantly higher than prebypass levels. Moreover, there were no significant differences in postoperative clinical characters, except for the intubated time. Conclusion In infants with congenital heart defects, pulmonary perfusion with hypothermic HTK solution during cardiopulmonary bypass could ameliorate lung function and reduce the inflammatory response.     

烧伤、冲击伤及烧冲复合伤后大鼠早期血清中性粒细胞弹性蛋白酶的变化及意义

Objective To explore the early changes in serum neutrophil elastase(NE) in rats with burn.blast injury or combined bum-blast injury and its significance.Methors A total of 176 male Spragne Dawley rats were randomly divided into four groups:control(C),burn(BU),blast injury(BL) and burn-blast combined injury(BB).Rats in C group were not injured.Animals in BU group were subjected to 25% TBSA full-thickness burn on back with 94℃ water for 12 seconds;Animals in BL group were inflicted with moderate blast injury with 5g 8701 compressed dynamite stick as the explosion source 75 cm away while left chest facing the explosive source;Rats in BB group were burned immediately after the blot injury similarly as in BL group.During the first 24 h post-injury,animals in BU and BB groups received intraperitoneal bronchoalveolar lavage fluid(BALF),water content of lung tissue and NE content in serum were determined at 0 h(C group),3 h,6 h,12 h,1 d,2 d,3 d,7 d post-injury.Results Protein concentration in BALF,water content of lung tissue and NE content in serum in SD rats of the injured groups were significantly higher than those in C group(P＜0.01 or P＜0.05),peaked within 2 d post-injury,especially at 2 d post-injury(NE content in serum:BU group,319.85±19.50,ng/ml;BL group,467.43±31.64 ng/ml;BB group,626.00±26.38 ng/ml vs.C group,78.53±25.10 ng/ml).Overall,protein concentration in BALF.water content of lung tissue and NE content in serum in BB group were significantly higher than BU and BL groups(P＜0.01 or P＜0.05).Correlation analysis showed that within 3 d post-injury.a significant positive correlation was found between the protein concentration in BALF,water content of lung tissue and NE content in serum(r=0.7910,0.8078,P＜0.05) in BU group.NE content in serum and protein concentration in BALF were significantly positively correlated in BB group(r=0.8672,P＜0.05).Conclusion NE may play an important role in early lung injury of burn or blast injury,especially in combined bum-blast injury.     

Calorie control increased vaspin levels of serum and periepididymal adipose tissue in diet-induced obese rats in association with serum free fatty acid and tumor necrosis factor alpha

Background Vaspin was recently identified as a novel adipokine that is predominantly secreted from adipose tissue and exerts insulin-sensitizing effects. This study was undertaken to elucidate the regulative effects of calorie control on the expression of vaspin and its potential mechanism.Methods Diet-induced obese Sprague Dawley （SD） rats were adopted as experimental models and accepted interventions of various ingestions and pioglitazone. Various differentiated stages of cultured 3T3-L1 cells were dealt with pioglitazone or TNFα in vitro for 48 hours to further verify findings in animal experiments.Results The rats were successfully induced into an obese experimental model with hyperinsulinemia, hyperlipidemia, and increased serum free fatty acid and TNFa by 12-week high-fat diet. It was found that depending on whether the rats were fed by a high-fat diet or a basal diet, there was extremely higher vaspin in the periepididymal fat pad than in subcutaneous adipose tissues by 16 weeks. Vaspin in sera and the periepididymal fat pad was much lower in rats with a high-fat diet than those with a basal diet （all P 〈0.05）, but vaspin in subcutaneous fat tissues was prone to increase in rats with a high-fat diet. A 4-week calorie restriction or pioglitazone on the obese rats resulted in a partial recovery of vaspin levels in sera and periepididymal adipose tissues, especially the latter revealed a more obvious superiority and increased vaspin levels of subcutaneous adipose. Surprisingly, the treatment of 4-week high-fat diet on non-obese rats did not significantly depress vaspin of sera and periepididymal adipose tissues. However, it is unknown if re-feeding generated the effect on vaspin levels of obese and non-obese rats on sera or adipose tissues. The correlation analysis showed that vaspin levels of serum and periepididymal fat tissues were negatively correlated with serum FFA, TNFα and insulin; meanwhile, there was a positive correlation between serum vaspin and vaspin of periepididymal fat tissues. Pioglitazone enhanced vaspin levels in cultured 3T3-L1 cells and supernatant in various differentiated stages, and this effect became more and more obvious along with the change of preadipocytes into mature fat cells. Administration of TNFα caused suppression on vaspin expression in differentiated stages of 3T3-L1 cells.Conclusions The present data indicated that a long-term high-fat diet could induce obesity metabolic syndrome in SD rats and finally lead to lower vaspin of sera and periepididymal fat, while pioglitazone and chronic calorie-control ingestion could enhance the production of vaspin. It was undoubtedly demonstrated that vaspin expression was strongly associated with insulin sensitivity, serum FFA, and TNFα.     

Bone marrow-derived mesenchymal stem cells protect rats from endotoxin-induced acute lung injury

Background  Acute lung injury (ALI) is a serious and common condition for which there are currently no specific strategies for treatment. Recent studies have suggested that bone marrow-derived multipotent mesenchymal stem cells (MSCs) may have therapeutic applications in multiple clinical disorders. We explored the biological effects of MSCs during endotoxin-induced ALI and the mechanisms involved.

Methods  MSCs were isolated from male rat bone marrow and the ALI model was induced by intravenous endotoxin injection. Female rats were sacrificed at 6 hours, 24 hours, 4 days, 1 week and 3 weeks post-injection of MSCs or saline and the lung tissue, bronchoalveolar lavage fluid, and serum were harvested for analysis. We further evaluated the survival of the rats and examined the effects of endotoxin-induced injury on the interaction between alveolar macrophages (AMs) and MSCs in ex vivo.

Results  There was a significant decrease in numbers of neutrophils in bronchoalveolar lavage fluid (P <0.05), and myeloperoxidase activity in the lung (P <0.01), and of TNF-α and IL-1β in serum (P <0.05) in the MSC treated rats at 4 days. Furthermore, MSC treated rats exhibited improved survival, lower lung injury score, higher concentration of IL-10 in the serum and a reduced hydroxyproline content, but these differences were not statistically significant. Moreover, co-cultures of MSCs and AMs had significantly reduced levels of TNF-α, IL-1β and macrophage inflammatory protein (MIP)-1α and significantly increased levels of IL-10 (P <0.05) in the culture supernatants.

Conclusions  Treatment with intravenous injection of bone marrow-derived MSCs have beneficial effects on endotoxin-induced ALI in rats. The beneficial effect might be achieved through the engraftment of differentiated MSCs in the lungs and appears derive more from their capacity to secrete soluble factors that modulate immune responses.

     

Influence of catgut implantation at acupoints on leptin and insulin resistance in simple obesity rats

OBJECTIVES:To investigate the influence of catgut implantation at acupoints on leptin resistance(LR) and insulin resistance(IR) in the simple obesity rat.METHODS:Rats were made obese with high-fat diets,after which surgical catgut was implanted at Zusanli(ST 36) and Neiting(ST 44) acupoints once a week for 4 weeks(implantation group).Rats from the implantation group were compared with normal rats and unoperated obese rats(control group).Gene expression of the leptin receptor(OB-R) was evaluated using RT-PCR and northern blot.Serum and hypothalamus leptin and insulin(INS) levels were determined by radioimmunoassay.Body weight,Lee’s index,body fat,serum and hypothalamus leptin and INS levels,and hypothalamic OB-R gene expression were determined before and after treatment.RESULTS:Body weights,Lee’s index,body fat,and serum leptin and INS levels were significantly higher in obese than in normal rats.Hypothalamic leptin and INS levels and OB-R gene expression were significantly lower in obese rats.Catgut implantation at acupoint promoted weight loss and decreased serum leptin and INS levels.Hypothalamic leptin and INS levels and OB-R gene expression increased significantly.CONCLUSIONS:Catgut implantation at acupoint adjusts central and peripheral leptin and promotes hypothalamic OB-R gene expression.This may be an important method for regulation of LR,IR and abnormal endocrinology and metabolism.     

Protective effect of diallyl trisulfide on liver in rats with sepsis and the mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production o