Effect of intestinal lymphatic pathway on apoptosis of lung tissue in rats by two-hit

AIM: To observe the effects of mesenteric lymph duct ligation on apoptosis of lung tissue,correlated gene expression with apoptosis and TNF-α,IL-6 contents in rats by two-hit.METHODS: 45 Wistar rats were divided into three groups: the ligation group,the non-ligation group and sham group,and the two-hit model was established by means of hemorrhage and LPS treatments.Ligating mesenteric lymph duct was conducted in ligation group.After 24 hours,the pathological sections of lung tissue were prepared for determining the apoptosis rate by TUNEL method and expressions of Bcl-2 and Bax were observed by immunohistochemical test.The lung homogenate was also prepared for determining the contents of TNF-α and IL-6 by ELISA.RESULTS: After two-hit,the apoptosis rate,Bax expression in lung tissue and contents of TNF-α and IL-6 in serum and lung homogenate in non-ligation group were increased and Bcl-2 expression was lower than that in sham group and ligation group significantly (P<0.01,P<0.05).Apoptosis rate in ligation group was no statistics difference with sham group (P>0.05),and the expression of Bcl-2 protein was increased and Bax was lower than that of sham group (P<0.01,P<0.05).CONCLUSION: Blockage of intestinal lymphatic pathway reduces the apoptosis of lung in two-hit rats,and its mechanism might relate to reduced the levels of TNF-α and IL-6 and improved the expression of Bcl-2 protein in lung by the ligation of mesenteric lymph duct.The mesenteric lymph of two-hit might play an important role in the pathogenesis of acute lung injury (ALI).     

Alteration of AQP2 expression in kidney tissues of emphysema model rats

AIM: To observe the expressions of aquaporin 2 (AQP2) in kidney tissues and the contents of endotoxin (ET), interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α) in serum in emphysema model rats, and to investigate the relationship between lungs and kidney in humoral metabolism. METHODS: The rats of emphysema were treated by injecting lipopolysaccharide into the trachea with cigarette smoking. Immunohistochemistry and Western blotting analysis were used to observe the expression of AQP2 in kidney tissues. RT-PCR was applied to detect the expression of AQP2 mRNA in kidney tissues. Blood sample and lung tissue were taken and the levels of ET, IL-1β and TNF-α were measured by radioimmunoassay. RESULTS: AQP2 expression in the kidney tissue in model group was greater than that in control group, and the expression of AQP2 mRNA showed the same results (P<0.01). ET, IL-1β and TNF-α levels in serum and lung tissue in model group were markedly higher than those in control group (P<0.01). CONCLUSION: In the emphysema model rats, AQP2 expression is up-regulated in the kidney tissue. The mechanism of emphysema may be related to increasing the levels of ET, IL-1β and TNF-α in the serum and lung tissue obviously.     

Mechanisms for a decrease in mortality of LPS-challenged mice by rhynchophylline

AIM: To observe effect of rhynchophylline （Rhy） on mortality and organ injury in endotoxemic mice and further investigate the mechanisms of its actions. METHODS: Male mice were randomly assigned into control, LPS, Rhy +LPS and Rhy group, and injected subcutaneously with normal saline (0.05 mL/10 g), or rhynchophylline once a day for 3 d, 1 h after subcutaneously treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally. Survival rate was recorded every 12 h for 6 d. In another experiment, 12 h after LPS injection, the left lung and intestine tissue sections were prepared for histological analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D),the serum was collected to detect the concentrations of alanine aminotransferase（ALT）, aspartate aminotransferase (AST ), bloodureanitrogen (BUN) and creatinine (Cr). In addition, the concentrations of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and interleukin-10 (IL-10) in serum at 2 h after LPS challenge were detected by enzyme-linked immunosorbent assay. The concentration of NO in serum at 8 h was detected by enzymic method. The effect of Rhy on survival rate of mice subjected to cecal ligation and puncture (CLP) was also observed. RESULTS: Mortality of mice challenged with LPS alone was higher significantly than that in control at 24 h after LPS challenge, pretreated with Rhy at a dose of 8 or 16 mg/kg increased markedly the survival rate of LPS-challenged mice. However, Rhy at a dose of 8 mg/kg significantly increased mortality of mice subjected to CLP. In the histological analysis, severe inflammation was observed both in the lung and intestine tissues in the LPS group. LPS elevated lung W/D, the levels of ALT, AST, BUN, Cr, TNF-α, IL-1β, IL-10 and NO in serum. Pretreatment with Rhy had no obvious improvement in the lung and intestine tissue injury, and no significant depression in the lung W/D and the serum levels of ALT, AST, BUN, Cr, IL-1β, IL-10 and NO, but decreased the level of TNF-α in serum significantly in LPS -treated mice. CONCLUSION: Pretreatment with Rhy reduces the mortality in endotoxemic mice, but not decrease the mortality of mice challenged with CLP, at least in part, through inhibiting the synthesis and secretion of TNF-α.     

Immunoregulatory effect of Dachengqi decoction on endotoxic ARDS in rats

AIM: To investigate the effect of Dachengqi decoction (DD) on the acute respiratory distress syndrome (ARDS) in rat model induced by endotoxin “two-hit”. METHODS: 48 healthy male Wistar rats were randomly divided into 4 groups: control group, ARDS model group, ARDS+DD treatment group and ARDS+dexamethasone treatment group (12 in each group). E. coli lipopolysaccharide (LPS) “two-hit” induced ARDS model in rats was established. The arterial blood gas analysis, lung wet/dry weight (W/D) ratio and lung tissue pathology observation and scoring were measured to evaluate the pharmacological effects of DD on ARDS. The levels of TNF-α, IL-1 and IL-10 were determined by ELISA to explore the immune regulatory mechanism of DD. RESULTS: (1) Treatment with DD significantly improved blood pressure in rats, increased oxygen saturation, decreased lung wet/dry weight ratio and lung injury score, relieved pulmonary edema and the inflammatory responses. (2) DD suppressed the productions of systemic and pulmonary pro-inflammatory mediators and promoted the release of anti-inflammatory mediators in lung. CONCLUSION: The inhibitory effects of DD treatment on pulmonary inflammatory response are not only related to reducing the extent of systemic inflammatory response, but also promote pulmonary anti-inflammatory production and regulate the balance of pulmonary pro-inflammatory mediators and anti-inflammatory cytokines. This specific regulatory effect protects the target organ from excessive inflammatory organ injury, discloses that DD has advantages in the treatment of endotoxic ARDS.     

Glucocorticoid attenuates LPS-induced acute lung injury by inhibiting p38MAP kinase in rats

AIM: To examine the role of glucocorticoid receptor (GR) in regulation of lipopolysaccharide (LPS)-induced lung injury. METHODS:Male Sprague-Dawley rats were divided into six groups randomly: control group (n=6), LPS group (n=6 each), Dex+LPS group (n=6 each), RU486 group (n=6), RU486+LPS group (n=6 each) and RU486+Dex+LPS group (n=6 each). All groups were subjected into 1 h, 3 h, 6 h and 12 h time point subgroups after LPS administration, except of control group and RU486 group. The concentrations of TNF-α and IL-6 in bronchoalveolar lavage fluids (BALF) were detected by ELISA. The histopathologic changes of lung tissues, the activation of p38MAPK and the expression of MKP-1 in lung tissue were also observed. Further, to confirm the role of GR in this model, the mortality of rats in LPS group vs RU486+LPS group and in Dex+LPS group vs RU486+Des+LPS group was compared. RESULTS: LPS induced lung injury and the secretions of TNF-α and IL-6 in BALF, which were significantly enhanced by pretreatment of RU486 (P<0.05). RU486 pretreatment also significantly increased the LPS-induced lethality (P<0.05). Dexamethasone attenuated LPS-induced lung damage, cytokine release and mortality rates, and the protective effects might be mediated by GR. Western blotting analysis showed dexamethasone inhibited the phosphorylation of p38MAPK in lung tissues by induction of MKP-1, and these actions were also GR dependent. CONCLUSION: GR plays an essential role in regulation of LPS-induced acute lung injury. Anti-inflammatory effects of hormone-activated GR may be mediated by inhibition of p38MAPK phosphorylation/activation, which is associated with the induction of MKP-1.     

Antagonistic effects of aminophylline on airway inflammation and oxidative lung tissue damage in chronic hypoxic rats

AIM: To investigate the effects of chronic hypoxia and antagonistic effects of aminophylline on airway inflammation and oxidative lung damage in rats. METHODS: Thirty-four male Wistar rats were randomly divided into three groups: normal control group (n=10); hypoxia group (n=12); aminophlline-treated group (n=12). The last two groups were both exposed to hypoxia 7 hours per day for 21 days. The third group was treated with aminophlline (100 mg·kg-1·d-1) before exposed to hypoxia. The level of tumor necrosis factor (TNF) -α, interleukin (IL)-10, lipid peroxide (LPO) and the activity of superoxide dismutase (SOD) were determined in blood and homogenates of lung tissue. RESULTS: Compared to the control group, the levels of TNF-α, IL-10 and LPO were significantly increased (P<0.01), the activity of SOD was significantly decreased (P<0.01) both in blood and homogenate of lung tissue. The ratio of TNF-α and IL-10 was significantly increased (P<0.01) in homogenate of lung tissue in hypoxia group. Compared to the hypoxia group, the levels of TNF-α, LPO and the ratio of TNF-α and IL-10 were significantly decreased (P<0.01) in blood and homogenate of lung tissue in aminophylline treated group, while the level of IL-10 and activity of SOD was significantly increased (P<0.05, P<0.01, respectively). CONCLUSIONS: Chronic hypoxia induces airway inflammation. Aminophylline produces anti-inflammatory effects on airway and anti-oxidantive effects on lung.     

Effect of the acupotome therapy and electro-acupuncture on the serum contents of IL-1β, IL-6 and TNF-α in rabbits with knee osteoarthritis

AIM: IL-1β, IL-6 and TNF-α play important roles in emergence of osteoarthritis. This study aims at observing the effect of the acupotome therapy and electro-acupuncture on the cytokines in serum of rabbits with osteoarthritis. METHODS: 52 New Zealand rabbits were divided randomly into the normal group, model group, acupotome group and electro-acupuncture group. Knee osteoarthritis of the model rabbits was made with the straightened immobilization method. The acupotome and electro-acupuncture therapies were applied for three weeks. One week after the treatment the serum was collected, and the changes of IL-1β, IL-6 and TNF-α in serum were detected with RIA. RESULTS: In comparison with the control group, the contents of IL-1β, IL-6 and TNF-α elevated significantly in the knee osteoarthritis model group (P<0.05). Compared to the knee osteoarthritis model group, the contents of the cytokines in acupotome treatment group and electro-acupuncture treatment group decreased significantly (P<0.05). No significant difference between the content of cytokines in the acupotome treatment or electro-acupuncture treatment groups (P<0.05) was observed, also no statistical difference between the acupotome treatment or electro-cupuncture treatment groups and the control group was found. However, the contents of the three cytokines in the acupotome treatment and electro-acupuncture treatment groups were still higher than those in control group. CONCLUSION: The acupotome and electro-acupuncture treatment can decrease the release of IL-1β, IL-6 and TNF-α, indicating that the two therapies play an important role in improvement of the articular cartilage cell injury and function through inhibiting the generation of matrix protease and alleviation of degradation of the cartilage matrix.     

Effect of rhIL-10 on IL-6 and TNF-α levels in serum and liver of lipopolysaccharide-challenged mice

AIM: To investigate the effect of recombinant human interleukin-10 (rhIL-10) on IL-6 and TNF-α levels in serum and liver of mice exposed to lipopolysaccharide (LPS). METHODS: rhIL-10 was prepared by using genetic engineering technology. Mice were intraperitoneally with 500 μg of LPS, and then were treated intravenously with various dosages of rhIL-10. The levels of IL-6 and TNF-α in hepatic tissue and serum were determined by ELISA at 12 h, 24 h, 48 h and 72 h post rhIL-10 treatment. RESULTS: rhIL-10 markedly inhibited the increase in IL-6 and TNF-α levels in hepatic tissue and serum at 12 h after rhIL-10 treatment in LPS-challenged mice, and the inhibition effect was significant at 24-48 h after rhIL-10 treatment （P＜0.05）. CONCLUSION: rhIL-10 can inhibit the increase in IL-6 and TNF-α levels induced by LPS in mice.     

Changes of pro-inflammatory cytokines in serum and lung of rats with oleic acid-induced acute respiratory distress syndrome and the effect of anisodamine

AIM: To observe the changes of interleukin-6(IL-6), IL-8 and tumor necrosis factor-α(TNF-α) in serum and lung at different time, and the effects of anisodamine (654-2) treatment in rats with oleic acid-induced ARDS. METHODS: The ARDS model induced by intravenous injection of oleic acid in the rat was used and levels of IL-6, IL-8, TNF-α in serum and lung tissue supernatant were measured using enzyme linked immunosorbent assay (ELISA). RESULTS: Levels of serum and lung tissue IL-6, IL-8, TNF-α in oleic acid type ARDS 4 h group were increased significantly. These cytokines in oleic acid type ARDS 8 h group were lower than that of ARDS 4 h group, but serum IL-6, TNF-α and lung tissue IL-6 were still higher than that of control group . In oleic acid type ARDS 16 h group, serum IL-6, TNF-α were lower than that of the ARDS 8 h group and serum TNF-α and lung tissue IL-6 were higher than that of control group. After 654-2 treatment, the levels of serum and lung tissue IL-6, TNF-α were decreased significantly. CONCLUSION: IL-6, IL-8 and TNF- α might play important roles in the oleic acid-induced ARDS in the rat. 654-2 might alleviate ARDS by inhibiting excess production of IL-6 and TNF-α.     

Effects of silymarin on LPS-induced acute lung injury in rats

AIM:To investigate the effects of silymarin on lipopolysaccharide (LPS)-induced acute lung injury in rats and its possible molecular mechanisms. METHODS:Fifty-eight male SD rats, weighting 230-250 g, were divided into four groups randomly: normal control (n=12); acute lung injury group (n=15), receiving intravenous LPS (O55∶〖KG-*2〗B5, 5 mg/kg); silymarin alone group (50 mg/kg, n=15); intervention group (n=16, receiving silymarin 50 mg/kg and LPS 5 mg/kg). The specimens were collected 6 hours later. The following changes, including blood gas analysis, the lung wet/dry weight ratio, the pulmonary vascular permeability, histological manifestations, lung tissue myeloperoxidase activity, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene in plasma and lung tissue, were observed. RESULTS:Compared with control group, the lungs of the rats in LPS treatment group showed significant hyperemia and spotted hemorrhage. The inflammatory granulocyte infiltrating, diffused alveolar septum thickening and spotted hemorrhage were observed in pathological examinations. The lung wet/dry weight ratio and Evans blue content (per gram) increased significantly after LPS treatment. The myeloperoxidase activity in plasma and lung tissue, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene were increased significantly in LPS treatment group. However, in intervention groups, all the above-mentioned measurements were reversed significantly by silymarin treatment compared with LPS treatment group. CONCLUSION:Silymarin may decrease inflammatory reaction and oxidative stress, and further decrease lung damage induced by LPS in rats, all indicating protection of silymarin against acute lung injury.     

Effects of strategies of heat-clearing and detoxifying,and Yin-invigorating and fluid-supplementing on HDL and ALP molecules associated with endotoxin-degrading

AIM: To explore the therapeutic strategies of heat-clearing and detoxifying (HCDT) and Yin-invigorating and fluid-supplementing (YIFS), the method of Chinese medicine, on the high density lipoprotein (HDL) and alkaline phosphatase (ALP) molecules, which are associated with endotoxin-degradation. METHODS: The animal model of endotoxemia was established by the injection of E. Coli lipopolysaccharides through rabbits ear vein. The endotoxemic rabbits were treated respectively with two representative herbal preparations of therapeutic principles against febrile diseases: HCDT or YIFS preparations. The serum levels of interleukin-1 (IL-1), tumor necrosis factor α (TNF-α) and HDL, the ALP activity, the expression of ALP mRNA in liver and kidney tissues were observed. RESULTS: The serum levels of IL-1 and TNF-α in HCDT group were significantly decreased, while the serum ALP activity and the expression of ALP mRNA in liver and kidney tissues were obviously increased, as compared with those in model group. Meanwhile, the serum levels of IL-1 and TNF-α in YIFS group were significantly reduced, and its plasma HDL level was elevated. CONCLUSIONS: Both the herbal preparations, HCDT and YIFS, have the scavenging effects on the overproduction of inflammatory cytokines such as IL-1 and TNF-α induced by endotoxin, but their effects on the endotoxin-degrading molecules might be different. HCDT principally increases ALP activity and enhances ALP expression in liver and kidney tissues, while YIFS might preferably facilitate the elevation of plasma HDL level.     

Experimental study on how brain-dead state affects the heart structure and function of Ba-Ma mini pigs and the mechanism

AIM:To investigate how brain-dead state affects the heart structure and function and the effect of PKC-α in BA-Ma mini pigs.METHODS:Ten Ba-Ma mini pigs were randomized into 2 groups: brain-dead group (n=5),and control group (n=5). The brain-dead model was made by increasing intracranial pressure,while the control group was maintained anesthesia for 24 h. The concentrations of cTnT,TNF-α,IL-1β and IL-6 in serum were determined at 6,12 and 24 h after brain death. At 24 h,heart tissues were observed by HE staining and electron microscope. The expression of PKC-α was detected by immunohistochemistry and RT-PCR.RESULTS:(1) Histological changes of myocardium: flaky bleeding under endocardium and dissolution of myocardium were found in optical microscope. In electron microscope dropsical mitochondria and confluent muscle fiber were found. (2) Changes of serum cTnT: serum cTnT for brain-dead group began to increase gradually since 6 h,and were significantly higher at each time point than those in control group (P<0.05). (3) Changes of inflammatory factors: IL-1β,IL-6,and TNF-α in brain-dead group began to increase gradually since 6 h,and were significantly higher at each time point than those in control group (P<0.05). (4) Changes of PKC-α expression: PKC-α mRNA and protein expressions in brain-dead group increased significantly at 24 h (P<0.05).CONCLUSION:Brain death may evoke heart structure and functional injury,and increase the levels of inflammatory factors and PKC-α. The activation of PKC-α may participate in the process of heart injury.     

Immunomodulatory effect of Sema4D on Th1/Th2 in asthmatic rats induced by RSV and intervention of Kechuanning oral liquid

AIM To explore the effect of semaphorin 4D (Sema4D) on the immune regulation of Th1/Th2 in asthmatic rats induced by respiratory syncytial virus (RSV) and the effect of Kechuanning (KCN) intervention on the asthmatic rats. METHODS Healthy SPF female Wistar rats (n=100) were randomly divided into 10 groups: normal group, model group, Sema4D group, Sema4D antibody group, low, middle and high doses of KCN groups, and Sema4D+low, middle and high doses of KCN groups. Except the rats in normal group, the other rats were treated with RSV combined with ovalbumin (OVA) to induce asthmatic model. The pathological changes of the lung tissue were observed by HE staining, and the levels of interleukin-4 (IL-4), IL-6, IL-8, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. RESULTS Inflammatory cell infiltration and inflammatory damage in lung tissues of the rats in model group, Sema4D group and Sema4D+low dose of KCN group were observed by HE staining, while these pathological changes were attenuated in Sema4D antibody group, low, middle and high doses of KCN groups, and Sema4D+middle and high doses of KCN groups. Compared with normal group, the levels of IL-4, IL-6, IL-8 and TNF-α in BALF of the rats in the other groups were significantly increased, and the level of IFN-γ was significantly lowered (P<0.05 or P<0.01). Compared with model group, the levels of IL-4, IL-6, IL-8 and TNF-α in Sema4D group were increased, and the content of IFN-γ was decreased (P<0.05 or P<0.01). Compared with model group, the levels of IL-4, IL-6, IL-8 and TNF-α in Sema4D antibody group, low, middle and high doses of KCN groups, and Sema4D+low, middel and high doses of KCN groups were significantly decreased (P<0.01), while the content of IFN-γ was significantly increased (P<0.05 or P<0.01). No significant difference among Sema4D antibody group, Sema4D+middle and high doses of KCN groups, and low, middle and high doses of KCN groups was observed (P>0.05). CONCLUSION Sema4D causes Th1/Th2 immune imbalance and aggravates asthma. Inhibition of Sema4D reduces the production of inflammatory factors and regulates the balance of Th1/Th2. KCN may attenuate RSV-induced immune inflammation of asthmatic rats by inhibiting Sema4D, so as to achieve the anti-asthma effect.     

Effects of diosmin on changes of TNF-α, IL-1β, IL-6, IL-8 and IL-10 in serum and kidney tissues of rats with kidney ischemia and reperfusion

AIM: To observe the effects of diosmin on the production of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6, IL-8 and IL-10 in serum and kidney tissues of rats with kidney ischemia and reperfusion (I/R). METHODS: Sprague-Dawley rats (180 in total) were randomly divided into 3 groups including sham operation group (sham),I/R group and diosmin+I/R group (diosmin+I/R). At the end of the experiment, the blood and kidney tissues were obtained and TNF-α, IL-1β, IL-6, IL-8 and IL-10 were detected by ELISA. RESULTS: The levels of TNF-α, IL-1β, IL-6, IL-8 and IL-10 in serum and kidney tissues in I/R group and diosmin+I/R group were significantly higher than those in sham group (P<0.01 or P<0.05). Following the development of the pathologic process, the level of TNF-α, IL-1β, IL-6 and IL-8 was significantly increased in I/R group and diosmin+I/R groups, but the level of IL-10 was significantly decreased in I/R group and significantly increased in diosmin+I/R group. The levels of TNF-α, IL-1β, IL-6 and IL-8 in I/R group was significantly higher than those in diosmin+I/R group (except TNF-α at 1 h in diosmin+I/R group). The level of IL-10 in diosmin+I/R group was significantly higher than that in I/R group (P<0.01 or P<0.05). CONCLUSION: Diosmin not only decreases the production of TNF-α, IL-1β, IL-6 and IL-8, but also promotes the production of anti-inflammatory cytokine IL-10, suggesting that the protective effect of diosmin on kidney I/R injury was associated with anti-inflammatory mechanism.     

Cytokine levels and the discrepancy of dexamethasone intervening in ALI rats induced by hydrochloric acid and lipopolysaccharide

AIM: To explore the cytokines level and the discrepancy of reaction to dexamethasone (Dex) in ALI rats induced by hydrochloric acid (HCl) and lipopolysaccharide (LPS). METHODS: Ninety-six SD rats were divided into six groups at random (n=16 in each group): NS group, HCl group, LPS group, NS+Dex group, HCl+Dex group and LPS+Dex group. Every group was divided into two subgroups: the bronchoalveolar lavage (BAL) subgroup and no bronchoalveolar lavage (NBAL) subgroup. The total leukocytes, PMN%, macrophage%, lymphocyte%, total protein in BALF and the wet/dry of the lung weight were measured. The concentrations of TNF-α, IL-1β, IL-4 and IL-10 in serum and BALF in every group were compared. RESULTS: (1) In the groups of LPS and HCl, the total leukocytes, PMN numbers, the protein concentration in the BALF and W/D were higher than those in control group (P<0.05). Compared to LPS groups, the percentage of macrophage increased in LPS+Dex group (P<0.05). (2) In serum and BALF of both LPS group and HCl group, the concentrations of TNF-α, IL-4 and IL-10 were higher than those in the control (P<0.01). The content of IL-1β in serum of all the groups was undetected. Compared to LPS groups, the concentrations of TNF-α and that of IL-1β decreased in LPS+Dex group (P<0.05). The concentration of IL-10 in LPS+Dex group was higher than that in LPS group (P<0.01). CONCLUSION: The permeability and the inflammatory cytokines in these two models were not consistent. Glucocorticoids play an effective role for resisting ALI induced by LPS but not HCl.     

Effects of thalidomide on the expression of NF-κB and TNF-α in experimental hepatic fibrotic rats

AIM: To study the effects of thalidomide on the expressions of nuclear factor κB (NF-κB) and tumor necrosis factor-α (TNF-α) in rat liver fibrosis.METHODS: The fibrosis of rat liver was induced by intraperitoneal injection of carbon tetrachloride thrice weekly.Meanwhile thalidomide (10 mg·kg^-1·d^-1 or 100 mg·kg^-1·d^-1) was given daily by the intragastric route for 8 weeks.Serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),prealbumin (PA),hyaluronic acid (HA) and laminin (LN),and hydroxyproline (HYP) contents in the liver,NF-κB p65 and α-smooth muscle actin (α-SMA) protein in the liver,IκBα and TNF-α protein in cytoplasm and NF-κB p65 protein in nucleus and TNF-α mRNA levels in the liver were studied.RESULTS: Compared with the model group,the Knodell score,serum ALT,AST,HA,LN levels and HYP contents in liver,NF-κB p65 protein in nucleus and α-SMA protein in the liver,and TNF-α mRNA and protein in the liver of rats given high dose of thalidomide were decreased significantly (P<0.01).Meanwhile PA level and IκBα protein in cytoplasm were elevated significantly (P<0.01).CONCLUSION: Thalidomide exerts its effect on the down-regulation of NF-κB-induced TNF-α via inhibiting dissociation and degradation of IκB and prevents liver fibrosis in rats.     

Effects of ganmao shuangjie heji on the inflammatory damage in the lungs of mice infected by IV FM1

AIM: To establish the influenza infected mouse model and study the anti-inflammatory effect of ganmao shuangjie heji. METHODS: IV FM1 infected mice were used as the animal model. The changes of pathology and the cytokine TNF-α, IFN-γ and IL-10 in the lung were observed by HE staining and ELISA (double antibody sandwich enzyme linked immunosorbent assay) after ganmao shuangjie heji treatment. RESULTS: After infected by influenza virus, severe interstitial pneumonia was induced in the model group. Mild interstitial pneumonia was observed in ganmao shuangjie heji treated group. The protein expressions of cytokine TNF-α, IFN-γ and IL-10 were higher in model group than those in the control group. The protein expressions of TNF-α and IFN-γ in ganmao shuangjie heji treated group decreased and IL-10 expression increased significantly compared with model group. CONCLUSION: Ganmao shuangjie heji decreases the expressions of TNF-α and IFN-γ, and increases the expression of IL-10, thus, alleviates inflammatory injury. The clinical application of this medicine can shorten the course of disease.     

Effects of neutrophil extracellular traps on acute lung injury in neonatal rats

AIMTo investigate the role of neutrophil extracellular traps (NETs) in neonatal rats with acute lung injury (ALI). METHODSThirty 7-day-old SD rats were randomly divided into normal saline control group, ALI group and ALI+deoxyribonuclease (Dnase) group (each n=10). The rats in ALI group were intraperitoneally injected with lipopolysaccharides (LPS) at 20 mg/kg, and the rats in ALI+Dnase group were intraperitoneally injected with Dnase at 5 mg/kg after LPS injection. After 6 h, the rats were anesthetized with chloral hydrate, bronchial alveolar lavage fluid (BALF) was collected, and the content of cell-free DNA (cf-DNA) in BALF was detected by fluorescence microarray. The right lung tissues were fixed in 4% paraformaldehyde, and the morphological structure of the lung tissues were observed by HE staining. The content of interleukin-6 (IL-6) and tumor necrosis factor-α （TNF-α） in the left lung homogenate was measured by ELISA. Immunofluorescence and Western blot were used to detect the production of citrullinated histone H3 (CitH3) and myeloperoxidase (MPO) in the rat lung tissues. RESULTSCompared with control group, the levels of cf-DNA, CitH3, MPO, IL-6 and TNF-α in lung tissues of neonatal rats in ALI group and ALI+Dnase group were all increased (P<0.05), and severe inflammatory infiltration in the lung tissues was observed. Compared with ALI group, the levels of cf-DNA, CitH3, MPO, IL-6 and TNF-α in ALI+Dnase group were decreased (P<0.05), and the inflammatory infiltration was attenuated. CONCLUSION In neonatal rats with ALI, the level of NETs is an important indicator of lung tissue injury, and NETs may be a new target for the treatment of neonatal ALI.     

Curcumin attenuates lipopolysaccharide/D-galactosamine-induced acute rat liver injury via activating autophagy of hepatocytes

AIMTo investigate the effect of curcumin (CUR) on autophagy of hepatovyte in rats with lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced acute liver injury (AHI). METHODSThe healthy male Sprague-Dawley (SD) rats were randomized into the control group,AHI group,CUR group, 3-methy-ladenine (3-MA) group and 3-MA+CUR group, with 6 rats in each group. AHI was induced with an intraperitoneal injection of LPS and D-GalN. Liver function was tested 12 h after LPS/D-GalN treatment. Pathological changes of liver tissues were analyzed by HE staining.The amount of autophagic bodies were observed by transmission electron microscopy. The protein levels of autophagy related-proteins LC3 and beclin 1 in livers were detected by Western blot. ELISA were used to examine the serum levels of tumor necrosis factor-α (TNF-α). RESULTSCompared with control group, the serum level of alanine aminotransferase (ALT) and asparated aminotransferase (AST) were significantly increased, hepatic pathological damage were aggravated and serum TNF-α level was significantly increased in AHI group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were increased (P<0.01). Compared with AHI group, the serum level of ALT and AST were significantly decreased, hepatic pathological damage were attenuated and serum TNF-α level was significantly reduced (P?<0.05), while the autophagic bodies and the protein levels of LC3 and beclin 1 were significantly increased in CUR group (P<0.01). Compared with CUR group, the serum level of ALT and AST were significantly increased, hepatic pathological damage were aggravated and serum level of TNF-α was significantly increased in 3-MA group and 3-MA+CUR group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were decreased (P<0.01). CONCLUSION Curcumin protects rats against LPS/D-GalN-induced liver injury, partially due to activation of hepatocyte autophagy in livers.