【Abstract】ObjectiveTo investigate the effect of Salvia Miltiorrhiza (SM) and Shengmai injection （SI） in treating systemic inflammatory response syndrome (SIRS) and their mechanism. Methods The animal model of SIRS was established by injectinglipopolysaccharide(LPS, 1 mg/kg)intraperitoneally. Forty Wistar rats were randomly divided into four groups: control group, SM group, SI group and combined treatment group (SM+SI group), which were treated with normal saline（5 ml/kg） plus LPS（1 mg/kg）, SM（5 ml/kg）plus LPSKG4（1 mg/kg）, SI（5 ml/kg）plus LPS（1 mg/kg）, SM（2.5 ml/kg） plus SI（2.5 ml/kg） and LPS（1 mg/kg） respectively. Six rats of each group were sacrificed for sample collection of blood, liver, lung and kidney 8 hours after LPS injection. Blood routine, serum TNF-α and IL-6 were measured. Specimen of organs were fixed in formalin and sent for routine pathological examination. The survival of other 4 rats of each group were observed untill 48 hours after LPS injection. SPSS 10.0 was used in statistical analysis. Results Two rats in control group died 13 hours and 22 hours after LPS injection respectively, the remaining 2 rats in this group and the rats in other 3 groups survived 48 hours after LPS injection. The white blood cell count of control group was significantly higher than that of other groups. The serum TNF-α and IL-6 of control group were significantly more than those of other groups. Pathological damages were found in all groups, and the most severe ones were in control group. SM and SI could decrease the level of serum TNF-α and IL-6 in the process of LPS-stimulated SIRS, down-regulate the severe inflammatory response, attenuate organ damages of the liver, lung and kidney, and increase forty-eihgt-hour survival rate obviously. Conclusion The experiment provides a theoretical base for clinical use of SM and SI in treatment of SIRS.
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 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.
Objective To investigate the impacts of cytokines （interleukin-4，IL-4；tumor necrosis factor-α，TNF-α） and medications of bronchial asthma （dexamethasone，aminophylline，salbutamol） on the activity of histamine N-methyltransferase（HMT） in tracheal epithelial cells.Methods BEAS-2B bronchial epithelial cells were cultured and treated with different concentration of TNF-α, IL-4, dexamethasone, salbutamol and aminophylline respectively. The activity of HMT in BEAS-2B cells was determined by high performance liquid chromatography.Results The activity of HMT in tracheal epithelial cells was (50±7) pmol?min-1?mg pro-1.TNF-α and IL-4 lowered the activity of HMT significantly at the concentration equal to or higher than 1 ng/mL and 5 ng/mL respectively，and reached the maximum inhibitory effect at the level of 10 ng/mL.Dexamethasone and aminophylline could ameliorate distinctly the inhibitory effect of TNF-α on the activity of HMT, while salbutamol had no significant inhibitory effect.Conclusions TNF-α and IL-4 exert the lowering effect on the activity of HMT，which would be one important cause of airway hyperreactivity.Glucocorticoids and theophyllines are administered to treat asthma partly due to its relieving mechanism of TNF-α negative effects on HMT.
Objective To comprehensively evaluate the association between TNF-α gene −308 G/A polymorphism and the risk of prostate cancer. Methods A meta-analysis was performed to analyze the association between −308 G/A polymorphism and the risk of prostate cancer risk. Results A total of 11 case-control studies (4 919 cases and 5 210 controls) were included in this meta-analysis. The result showed no statistically significant differences in all genotype distribution between prostate cancer cases and controls: dominant model (OR=1.11, 95%CI 0.90 to 1.36, P=0.33), recessive model (OR=0.91, 95%CI 0.70 to 1.18, P=0.47), GA versus GG (OR=1.11, 95%CI 0.90 to 1.37, P=0.33), AA versus GG (OR=0.92, 95%CI 0.71 to 1.20, P=0.55), A versus G (OR=1.07, 95%CI 0.91 to 1.26, P=0.39). In the subgroup analysis by ethnicity, no statistically differences were found between prostate cancer cases and controls. Conclusion This results of meta-analysis suggests that TNF-α gene –308G/A polymorphism may not be a risk factor of prostate cancer. Due to the limited quantity of the includied studies, further studies are needed to validate the above conclusion.
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.
ObjectiveTo explore the therapeutic effect and its possible mechanisms of somatostatin combined with antibiotics on acute cholecystitis through the detection of serum tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) in rabbits. MethodsForty-five rabbits were randomly averagely classified into three groups following the establishment of acute cholecystitis model: control group, blank group， and experimental group. The rabbits in control group received cefazolin sodium and metronidazoie by intravenous injection twice a day. The rabbits in experimental group got a hypodermic injection of somatostin (20 μg/kg) twice a day besides antibiotics, while these drugs were replaced by equal volume of normal saline for the rabbits in control group. The concentrations of serum TNFα and CRP were detected by enzyme-linked immunosorbent assay and histomorphological and electron microscopic changes of gallbladder in rabbits were observed on 3 d after administer. ResultsThe concentrations of serum TNF-α of rabbits in experimental group 〔(401.6±48.7) pg/ml〕 were significantly lower than those in control group 〔(767.3±67.4) pg/ml〕 and blank group 〔(806.7±61.2) pg/ml〕, P=0.000 and P=0.000, while the difference between the latter two groups was not significant (P=0.196). The concentrations of serum CRP of rabbits in experimental group 〔(16.2±1.1) mg/L〕 were significantly lower than those in control group 〔(55.4±1.2) mg/L〕 and blank group 〔(72.8±8.9) mg/L〕, P=0.000 and P=0.000, and which was higher in blank group compared with control group (P=0.018). The Histopathological results showed that gallbladder wall emerged mulifocality mucosal fluid necrosis, lamina propia hyperemia, bulk neutrophil infiltration and sequent alleviation of reaction in the rabbits of experimental group when compared with the rabbits of blank group and control group. Electron microscopic results demonstrated that the intercellular junction of gallbladder kept relative integrity and the swelling and vacuolar degeneration of mitochondria and endoplasmic reticulum obviously relieved. ConclusionsSomatostatin can significantly reduce the concentrations of serum TNF-α and CRP in the model of rabbits acute cholecystitis, which may protect the mucous membrane of gallbladder from the inflammation reaction.
【Abstract】Objective To study the influence of early hemofiltration on plasma concentrations of proinflammatory cytokines TNF-α and IL-1β and their transcription levels in severe acute pancreatitis (SAP) pigs. Methods The model of SAP was induced by retrograde injection of artificial bile into pancreatic duct in pigs. Animals were divided randomly into two groups: SAP hemofiltration treatment group (HF group, n=8) and SAP no hemofiltration treatment group (NHF group, n=8). TNF-α and IL-1β plasma concentrations were measured by ELISA. Their transcription levels in the tissues of pancreas, liver and lung were assayed by semi-quantitative reverse transcription polymerase chain reaction. Results After hemofiltration treatment, the plasma concentrations of TNF-α and IL-1β increased gradually but were lower than those of NHF group at the same time spot 〔at 6 h after hemofiltration treatment, (618±276) pg/ml vs (1 375±334) pg/ml and (445±141) pg/ml vs (965±265) pg/ml, P＜0.01〕. At 6 h after hemofiltration treatment, the transcription levels of TNF-α and IL-1β in tissues of pancreas, liver and lung were lower than in NHF group (57.8±8.9 vs 85.7±17.4, 48.0±8.1 vs 78.1±10.2, 46.2±9.6 vs 82.4±10.5; 55.9±9.0 vs 82.2±15.7, 40.6±9.2 vs 60.0±10.6, 35.7±9.8 vs 58.1±9.3, P＜0.01). Conclusion Early hemofiltration can reduce TNF-α and IL-1β plasma concentrations and transcription levels in SAP pigs.
Objective To investigate the clinical significance of insulin resistance ( IR) in chronic obstructive pulmonary disease ( COPD) .Methods Patients with stable COPD were recruited while healthy volunteers were enrolled as control. The diagnosis and severity assessment were made according to chronic obstructive pulmonary disease diagnosis and treatment guideline ( revised edition 2007) . Fasting serum levels of glucose ( FBG) , insulin ( FIN) , blood lipids, fibrinogen, C-reactive protein ( CRP) , tumor necrosis factor ( TNF-α) , and interleukin-6 ( IL-6) were measured. The degree of IR was calculated by IAI( IAI =1/FBG ×FIN) . The relationship of IR with COPD severity and above parameters was analyzed. Results A total of 121 subjects with COPD were enrolled in which 22 cases of mild COPD, 28 cases of moderate COPD,34 cases of severe COPD, and 37 cases of extremely severe COPD. The levels of FBG and FIN were significantly higher in the COPD group than those in the normal control group ( P lt;0. 05) . ISI in the COPD patients was higher than that in the controls ( - 3. 88 ±0. 54 vs. - 3. 40 ±0. 28, P lt;0. 05) . The levels of CRP, fibrinogen, TNF-α, and IL-6 were significantly higher in the COPD group than those in the control group ( P lt;0. 05) . The levels of CRP, TNF-αand IL-6 increased progressively with the severity of COPD. There was a negative correlation between ISI and the severity of COPD ( r = - 0. 512, P lt; 0. 01) , positive correlations of CRP, fibrinogen, TNF-αand IL-6 levels with COPD severity, respectively( r=0. 710, 0. 600,0. 708,0. 707, all P lt;0. 01) , and negative correlations of ISI with the levels of CRP, fibrinogen, TNF-α and IL-6 ( r = - 0. 384, - 0. 240, - 0. 298, - 0. 396, all P lt; 0. 01) , respectively. Conclusion There is an increase in fasting serum insulin and insulin resistance in patients with COPD compared with healthy subjects, which deteriorates with severity of COPD.
ObjectiveTo establish 16HBE cell lines stably expressing glutathione S-transferase mu 5 (GSTM5) gene, and explore the mechanism of GSTM5 nuclear translocation. MethodsRecombinant lentiviral expression vector containing GSTM5 gene was constructed and lentivirus was produced. After lentivirus infection of 16HBE cells, 16HBE-GSTM5 cell lines were obtained by screening with puromycin. Expression of GSTM5 in different cells was examined by RT-qPCR and Western blot. The nuclear translocation of GSTM5 was observed by confocal laser scanning microscope, after the 16HBE-GSTM5 cell lines were treated with tumor necrosis factor-α (TNF-α; 10 ng/ml) for 0.5 hour. ResultsLentiviral expression plasmids, PLVX-puro-3*flag-SBP-GSTM5-C and PLVX-puro-GSTM5-SBP-3*flag-N, were constructed and lentiviral particles were successfully packed. After infected with lentivirus and screened by puromycin, two cell lines, 16HBE-GSTM5-SBP-3*flag-N and 16HBE-3*flag-SBP-GSTM5-C, were obtained. GSTM5 expression in these two cell lines was significantly higher compared with the control group and parental cells. After treated with TNF-α for 0.5 hour, the nuclear translocation of GSTM5 in 16HBE-GSTM5-SBP-3*flag-N was much more obviously than that in 16HBE-3*flag-SBP-GSTM5-C. ConclusionThe N-terminal region of GSTM5 is critical for nuclear translocation induced by TNF-α, which is mediated by a novel and non-classical nuclear localization signal.