Objective To observe the serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), platelet 5-HT and blood platelet count, emotion and burn injury healing of patients with moderate and severe burn injury and anxiety-depression symptoms. Methods In-patients with moderate and severe burn injury were selected from 2003.4 to 2005.2 and then divided into anxiety-depression group and control group according to their anxiety-depression scores by Hamilton Rating Scale for Depression (HAMD ) and Hamilton Rating Scale for Anxiety (HAMA) 3 days after being burnt. Routine therapy was given to two groups, which lasted 1 month. Their scores of anxiety and depression and the degree of injury healing were observed, and the serum levels of TNF-α and IL-6, platelet 5-HT and blood platelet count were measured in the two groups. Results Fifty-one in-patients with moderate and severe burn injury were divided into the anxiety-depression group (24 cases) and the control group (27 cases). After 30-day treatment, the depression scores did not decrease in the anxiety-depression group (P=0.12), but the anxiety scores decreased (P=0.00). In the anxiety-depression group, the burn injury healing time was postponed (P=0.00), the serum levels of TNF-α increased (P=0.00), and the platelet 5-HT levels decreased (P=0.04) before and after treatment. Conclusion Depressive reaction occurs in patients with moderate and severe burn injury, which is a continuously negative emotion. It can lead to high levels of serum TNF-α, reduction in platelet 5-HT, and delayed burn injury healing time. Due to the limited sample size and different location of patients, there may be some bias in this conclusion. We are prepared to increase the sample size and select patients in the same region in further relevant studies.
ObjectiveTo investigate The role of tumor necrosis factor-α (TNF-α) in pancreatitis-associated adrenal cells' apoptosis of severe acute pancreatitis (SAP). MethodsForty Wistar rats were randomly divided into sham operation group (SO group) and SAP group by random number method, the SAP group was divided into 3, 6, 12, and 24 h 4 subgroups, 8 rats in each group. SAP model was induced by retrograde injection of 5% sodium taurocholate into the bilipancreatic duct. At 3, 6, 12, and 24 h after operation, serum amylase and lipase was measured, adrenal injury was evaluated by histological examination, apoptosis of the adrenal cells was observed by TUNEL method, and expressions of TNF-α and Caspase-3 protein were detected by Western blot. ResultsThe levels of serum AMY and LIP, histopathological scores of pancreatic tissues and adrenal tissues at each time point after operation in SAP group increased significantly than SO group (P < 0.05). With the duration extension of SAP, the apoptosis index of adrenal cells in SAP group progressively heightened, and were higher than those in the SO group (P < 0.05). And the expressions of TNF-α and Caspase-3 protein in adrenal tissues of SAP group gradually increased, at 24 h this data slightly decreased, but still higher than SO group (P < 0.05). ConclusionTNF-α may be involved in the pathogenesis of adrenal injury in SAP rats by activate the protein expression of Caspase-3.
【Abstract】ObjectiveTo investigate whether tumor necrosis factor-α (TNF-α) enhance the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9(MMP-9) in hepatic cancer cell line HepG2 or not. Methods Cultured HepG2 cells were treated by TNF-α with various concentration and time. The morphological changes of HepG2 cells were studied microscopically and the proliferation of HepG2 were detected by methyl thiazolyl tetrazolium (MTT). The expression of VEGF and MMP-9 mRNA in cultured HepG2 were determined by relative quantitative reverse transcription polymerase chain reaction. The VEGF and MMP-9 protein level in supernatants and in cytoplasm were determined by enzymelinked immunosorbent assay (ELISA) and by immunocytochemical staining, respectively.Results There was a little morphological changes in HepG2 with TNF-α treatment, but no change of cell proliferation in corresponding time. The expression of VEGF and MMP-9 mRNA was enhanced gradually with the TNF-α concentration increasing, the VEGF and MMP-9 protein level in supernatants and in cytoplasm was elevated gradually with the concentration increasing. There was a dependance on the concentration when the concentration of TNF-α was lower than or equal to 104 U/L. Furthermore, the effect of promotion was close to peak when the TNF-α concentration up to 104 U/L; but no timeeffect pattern observed. Conclusion TNF-αJP can enhance the expression of VEGF and MMP-9 at the level of mRNA and protein in hepatic cancer cell line.
ObjectiveTo investigate the inhibitory effects of L arginine (L arg) on systemic inflammatory response after cardiopulmonary bypass(CPB).MethodsFifty one patients with rheumatic heart disease were randomly divided into two groups: L arg group ( n =25) and control group ( n =26). For L arg group, L arg at 300mg/kg was given during operation. Plasma levels of tumor necrosis factor α(TNF α),interleukin 1β(IL 1β)and interleukin 10(IL 10) were measured by enzyme linked immunosorbent assay technique at baseline(before operation) and at 2,4,8,24 and 48 h after CPB termination.ResultsTNF α,IL 1β and IL 10 levels were increased in both groups after CPB ( P lt;0.05); levels of TNF α, IL 1β returned to normal at 48 h after CPB; In L arg group, TNF α and IL 1β levels were significantly lower than those in control group at 4,8 and 24 h after CPB ( P lt; 0 05). No significant difference were detected in IL 10 between groups( P gt;0.05).ConclusionL arg may decrease plasma levels of TNF α and IL 1β after CPB, it implies L arg may inhibit inflammation induced by CPB.
Objective To study the effect and intrinsic mechanism of acute suppurative peritonitis associated ascitic fluid (ASPAAF) on experimental liver injury of rats. Methods Thirty-two male or female Sprague-Dawley (SD) rats were randomly divided into two groups: ASPAAF group (n=16) and control group (n=16), in which 8 ml ASPAAF or normal saline (NS) were injected into the peritoneal cavity, respectively. The rats were killed at each time intervals after peritoneal cavity injection (6 h and 12 h) respectively in two groups and specimens were made to detect the levels of serum TNF-α, endotoxin and liver function (AST, ALT and STB). The level of TNF-α in liver tissues was measured. The pathological change of liver was observed by microscope. Results The levels of TNF-α, endotoxin, ALT, AST and STB in serum and the levels of TNF-α in liver tissues at different time points were markedly higher in ASPAAF group compared with those in control group (P＜0.05), and these indexes increased with increasing time in ASPAAF group (P＜0.05). In ASPAAF group, hepatic tissue appeared hydrops, even spotty necrosis and the changes at 6 h and 12 h were not obvious different. No abnormal pathological change of hepatic tissue was found in control group. Conclusion ASPAAF can induce the injury of the liver in rats, which may involved in TNF-α and endotoxin.
Objective To study the effect of tumor necrosis factor-α( TNF-α) onhypermetabolism of skeletal muscle protein in rats with chronic obstructive pulmonary disease ( COPD) and explore its underlying mechanism. Methods Forty-five SD rats were randomly divided into a normal control group, a COPD group and a COPD + TNF-α group, with 15 rats in each group. COPD model was established by passive cigarette smoking in COPD group and COPD + TNF-αgroup. Then the extensor digitorium longus muscles ( EDL) were dissected and incubated in vitro muscle incubation system with adequate oxygen supply. The EDL were either cultured with or without recombinant rat TNF-α( 10 μg/L) . The mRNA and protein expressions of proteasome subunit C2 in EDL were quantified by real-time quantitative PCR and Western blot analysis, respectively. Results The mRNA and protein expressions of proteasome subunit C2 were both significant higher in the COPD group and COPD + TNF-αgroup than those in the normal control group( P lt;0. 01 or 0. 05) . The COPD+TNF-αgroup had higher expression of proteasome subunit C2 mRNA than that in the COPD group( P lt; 0. 01) , whereas the protein expression was not significantly different( P gt; 0. 05) . Conclusion Incresed proteolytic metabolism in skeletal muscle in COPD might be regulated by TNF-αactivated ubiquitin-dependent pathway.
Objective To investigate influence of genders on the activity of nuclear factor-kappa B (NF-κB) in lungs of endotoxemic rats. Methods Twenty female and 20 male Wistar rats were randomly divided into four groups as follow: female control group (n=10), male control group (n=10), male endotoxemic group (n=10), and female endotoxemic group (n=10). The endotoxemic rats model was made by injecting lipopolysaccharide (5 mg/kg) into the abdominal cavity. Tissue samples were collected from the lungs in different groups and electrophoresis mobility shift assay was used to measure the activity of NF-κB. The levels of serum TNF-α and estrogen were measured at the same time. Results There was no significant difference between the activities of NF-κB in male and female control groups (1.33±0.24 vs 1.47±0.40）, and there was also no significant difference between other items in these groups as well (Pgt;0.05). Yet, the activity of NF-κB (female: 12.10±2.89; male: 19.53±2.12) and the level of TNF-α 〔female: (4.10±0.72) ng/ml; male: (6.37±1.29) ng/ml〕 were significantly increased after injection of lipopolysaccharide (Plt;0.01), and the indices in female group were significantly lower than those in male group (Plt;0.01). Correlation analysis showed that there was a positive relation between the activity of NF-κB in lungs and the level of TNF-α (female: r=0.921 1, P=0.013; male: r=0.907 2, P=0.017), and there was a negative correlation between the activity of NF-κB and the level of estrogen (female: r=-0.887 5, P=0.017; male: r=0.872 3, P=0.022) in both male endotoxemic group and female endotoxemic group (Plt;0.05). Conclusion Gender may be one of the factors that influence the activity of NF-κB in the lungs of endotoxemic rats. While on the other hand, endogenous estrogen may protect the lungs of endotoxemic rats from injury by inhibiting the activity of NF-κB.
ObjectiveTo investigate the correlations between tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) with acute myocardial dysfunction after severe thoraco-abdominal injuries and possible mechanisms. MethodsClinical data of 82 patients with severe thoraco-abdominal injuries who were admitted to the 253rd Hospital of People's Liberation Army from January 2009 to June 2012 were retrospectively analyzed,whose trauma index (TI) were all above or equal to 17 points. Patients with concomitant brain injuries and patients who were brought in dead were excluded from this study. There were 58 male and 24 female patients with their age of 16-76 (43.59±16.33) years. There were 17 patients with open injuries and 65 patients with closed injuries. There were 23 patients with fall injuries,47 patients with traffic injuries,8 patients with blunt injuries,and 4 patients with penetrating injuries. The time from injury to admission was 1.51±0.52 hours. Blood creatine kinase-MB (CK-MB) cardiac troponin T (cTnT) TNF-α and LPS were examined during emergency treatment,and the correlations between the results were analyzed. ResultsMyocardial dysfunction was shown by CK-MB of 158.74±31.59 U/L and cTnT of 496.25±58.46 pg/ml. Injury factors were TNF-α of 36.41±18.09 ng/ml and LPS of 343.66±106.02 U/L. CK-MB was positively correlated with TNF-α and LPS with the correlation coefficient (r) of 0.923 1and 0.883 2 respectively. cTnT was also positively correlated with TNF-α and LPS with r of 0.955 6 and 0.889 1 respectively. ConclusionBoth TNF-α and LPS participate in the pathogenesis and development of acute myocardial dysfunction after severe thoraco-abdominal injuries. Early intervention against TNF-α and LPS may alleviate acute myocardial dysfunction and improve patients' survival rate after severe thoraco-abdominal injuries.
Objective To investigate the changes of expression of zonula occludens-1(ZO-1) in rats with severe acute pancreatitis (SAP), and to study the relationship between the ZO-1 protein and microvascular injury in rats with SAP. Methods Forty-eight Wistar rats were randomly divided into sham-operation (SO) group and SAP group, each group enrolled 24 rats. Pancreas of rats in SO group were flipped only after laparotomies, but rats of SAP group were injected with 5% sodium taurocholate by retrograding cholangiopancreatography micro pump to produce the SAP model. At 6, 12, and 24 hours after operation, 8 rats were sacrificed to get abdominal aortic blood for testing the levels of peripheral blood amylase, trypsin, interleukin-8(IL-8), tumor necrosis factor-α(TNF-α), and ZO-1 protein. At the same time, pancreatic tissues were got to perform HE staining and immunohistochemical staining for observation of the pathological changes and the expression of ZO-1 protein respectively. Results Compared with SO group at the same time, the levels of peripheral blood amylase, trypsin, IL-8, TNF-α, and ZO-1 protein were all higher in SAP group (P < 0.05). The level of amylase in SAP-24 hours group was higher than those of 6 hours group and 12 hours group(P < 0.05), the levels of trypsin, IL-8, and ZO-1 protein in SAP group increased over time (P < 0.05), but levels of TNF-αin 3 time points of SAP group did not differ with each other significantly(P > 0.05). Results of regression showed that in the SAP group, the level of ZO-1 protein in serum was significantly positive correlated with pathological score of pancreatic tissue(b=0.96, P < 0.05), levels of serum amylase(b=0.87, P < 0.05), trypsin(b=0.72, P < 0.05), and serum IL-8 (b=0.69, P < 0.05), but was not significantly correlated with level of TNF-α(P > 0.05). HE staining results showed that damage of pancreatic tissues became worse over time in SAP group, and the pathological score of SAP-6 hours group was lower than those of 12 hours group and 24 hours group (P < 0.05). Immunohistochemical staining results showed that, in SAP group, with the extension of time, the number of ZO-1 protein granules in pancreatic acinar cells and capillary wall reduced, and expressed in capillaries discontinuously. Conclusion During the course of SAP, the concentration of serum ZO-1 protein increase, but its expression in the pancreatic tissue degrade, which is closely associated with microvascular injury and progression of pancreatic tissues.
ObjectiveTo investigate the expression of tumor necrosis factor α(TNF-α ) in isolated rat heart at different time points after myocardial hypoxia/reoxygenation. MethodsThe isolated langendorff perfused rat heart model was established. Forty-eight SD rats were randomly divided into four groups: a sham group, hypoxia/reoxygenation groups including a H/R 0.5 h group, a 1 h group and a 2 h group. The heart rate(HR), the 1eft ventricular development pressure(LVDP), maximal rates of increase/decrease of the left ventricular pressure(±dp/dtmax) were continuously recorded. The concentrations of TNF-α and creatine kinase-MB(CK-MB) in myocardium, mRNA expression of TNF-α in myocardium were tested. Ultra structure of myocardium was observed under electron microscope. ResultsThe levels of LVDP, ±dp/dtmax, and HR of hypoxia/reoxygenation group were significantly lower than those in the sham group(P<0.05).The levels of TNF-α and CK-MB and the expressions of TNF-α at mRNA level in the hypoxia/reoxygenation group were higher than those in the sham group(P<0.05).There were significant differences in the above parameters among the H/R 0.5 h group, the 1 h group, the 2 h group(P<0.05).The concentrations of TNF-α and CK-MB, the mRNA expression of TNF-α were higher in the I/R 2 h group than those in the other two groups. ConclusionThe high expression of TNF-α in myocardium after myocardial hypoxia/reoxygenation in rats is related to the degree of myocardium damage and may lead to myocardial injury.