Objective To investigate the mRNA and protein expression of human β-defensin-2 (hBD-2) induced by lipopolysaccharide (LPS)，IL-1β and TNF-α in human airway primary epitheliums.Methods The bronchial primary epitheliums from human were stimulated with LPS，IL-1β and TNF-α respectively and then were harvested for hBD-2 expression detection.The mRNA expression of hBD-2 was detected by RT-PCR，and the protein expression by immunocytochemistry and western blot.Results There was a small expression of hBD-2 mRNA in human airway primary epitheliums before stimulation.The hBD-2 mRNA expression was significantly increased after 3 hours of LPS，IL-1β and TNF-α stimulation respectively and the expression increasement was in a dose dependent manner.The hBD-2 protein could be detected in cytoplasm after 4 hours of LPS (0.1 μg/mL)，IL-1β (1 ng/mL) and TNF-α (10 ng/mL) stimulation.Conclusions LPS and proinflammatory cytokines can induce the mRNA and protein expression of hBD-2 in a short time.The expression of hBD-2 may play an initial defense role against bacterial invasion.
Objective To study the effect of nontypeable Hemophilus influenzae(NTHi) strain ATCC49247 on proinflammatory cytokines expression of human A549 lung epithelial cell line. Methods Confluent A549 cells were co-incubated with NTHi, NTHi+Erythromycin(10 mg/L), NTHi+Gentamicin(100 mg/L), and NTHi+Dexamethasone(100 μmol/L),and nuclear factor kappa B(NF-κB) inhibitor primed cells were co-incubated with NTHi for 24 h. Then levels of interleukin-8（IL-8） and tumor necrosis factor-α(TNF-α) in the supernatant was assayed by enzyme-linked immunosorbent assay(ELISA) and the expression of intercellular adhesion molecule-1(ICAM-1) in cells was detected by immunohistochemistry staining. Results A549 cells were transformed and died after co-intubated with NTHi for 24 h. NTHi induced A549 cells to release significantly greater amounts of IL-8, which was inhibited by NF-κB inhibitor pyrrolidine dithiocarbamate（PDTC）. Incubating of A549 cells with NTHi significantly induced release of IL-8 and the expression of ICAM-1, which was blocked by erythromycin and dexamethasone and not by gentamicin. TNF-α was not detected in all circumstances. Conclusions NTHi can increase significantly the release and expression of proinflammatory cytokines through NF-κB pathway. Antibacterial drug erythromycin also has anti-inflammatory effect.
Objective To identify the effects of single immunoglobin IL-1 receptor related protein （SIGIRR） on inflammation induced by high mobility group box 1 （HMGB1） in A549 derived from human alveolar epithelial cells. Methods Eukaryotic expression vectors pCDNA3.1（+） constructed with SIGIRR cDNA were transiently transfected into A549 cells，in which SIGIRR was forced to be over-expressed. Western blot and RT-PCR were applied to detect the expression level of SIGIRR after transfection. After the stimulation by HMGB1，the transcriptional activity of NF-κB in A549 cells was detected by dual-luciferase reporter assay system，and the protein levels of inflammatory cytokine TNF-α and IL-1β were measured by ELISA. Results The expression level of SIGIRR increased significantly in A549 cells transfected with SIGIRR vectors. The transcriptional activity of NF-κB was enhanced obviously after HMGB1 treatment in A549 cells by dual-luciferase reporter assay system，while the transfection of SIGIRR vectors decreased the activity. The protein levels of TNF-α and IL-1β were down-regulated in A549 cells over-expressing SIGIRR after HMGB1 stimulation compared with the non-transfected cells. Conclusions Up-regulated SIGIRR expression can inhibit HMGB1-induced proinlammatory cytokine release in A549 cells such as TNF-α and IL-1β. The transcriptional activity of NF-κB is dampened by SIGIRR transfection，implying that the anti-inflammatory effects of SIGIRR may be involved in the regulation of NF-κB.
Abstract： Objective To investigate the acute cardioprotective effect of 17b-estradiol (17b-E2) against severe myocardial ischemia/reperfusion (I/R) injury in rabbits and the mechanism of the effect. Methods We established the model of myocardial I/R in vivo by occluding the left anterior descending coronary artery of the rabbits (who underwent coronary occlusion for 40 minutes followed by 3 hours of reperfusion). Twentyfour New Zealand white male rabbits were randomly divided into two groups with 12 in each group. Before coronary occlusion, 1 ml of ethanol or 17b-E2 at 10 μg/kg was administered intravenously to the rabbits in the control group and the experimental group respectively. The serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured by enzymelinked immunosorbent assay (ELISA) at the following time points: before occlusion, 40 minutes after occlusion, 1 hour, 2 hours and 3 hours after reperfusion. Activation of p38 mitogen activated protein kinase(MAPK) was determined by Western blotting analysis, and apoptosis of cardiocytes was identified by terminal deoxynucleotidlyl transferase mediated deoxyuridinebiotin dUTP Nick End Labeline (TdT)mediated dNTP nick end labeling (TUNEL) staining. Results During myocardial ischemia, TNF-α decreased significantly in the experimental group compared with the control group (F=0.007,P=0.001), while there was no difference in IL-6 between the two groups (F=0.616,P=0.095). During the process of reperfusion, the levels of TNF-α and IL-6 in the experimental group were significantly lower than those in the control group (Plt;0.01). Besides, the activation of p38 MAPK and apoptotic index for the experimental group were also lower (45.07%±2.73% vs. 61.25%±2.41%, t＝－15.398, P=0.000; 11.21%±3.85% vs. 22.02%±4.49%, t＝－6.332, P=0.000). Conclusion The cardioprotective effect of 17b-E2 against myocardial I/R may be attributed to its antiinflammatory and antiapoptotic properties, which is probably associated with the inhibition of 17bE2 on p38MAPK activity.
【摘要】 目的 观察胎羊宫内心脏介入手术胎羊血气及血浆炎性细胞因子的变化。方法 8只怀孕双胎山羊,双胎之一为实验组,在相同麻醉条件下,实验组进行胎羊心脏介入治疗,并抽取血样标本。监测胎羊的心率、血气、乳酸值,运用ELISA法检测治疗组及对照组胎羊白介素(IL)1、IL6、IL8及肿瘤坏死因子(TNFα)。结果 2只胎羊因手术中发生心包填塞死亡,存活的6只胎羊手术前pH值较手术后有明显下降(Plt;005),手术前后乳酸浓度上升(Plt;005),PCO2、PO2差异无统计学意义(Pgt;005),手术前血浆IL1、IL6、IL8的浓度较手术后高(Plt;005),手术前后TNFα的浓度变化无统计学意义(Pgt;005)。结论 胎羊宫内心脏介入手术可引起胎羊血浆pH值下降,乳酸浓度上升,及细胞因子IL1、IL6、IL8浓度上升。【Abstract】 Objective To observe the change of blood gas and inflammatory cytokines during intrauterine cardiac intervention surgery on the fetal lambs. Methods Eight pregnant goats with two fetal in each goat were included. With the same anesthesia condition, one of the twin fetus was chose to perform the intrauterine cardiac intervention surgery. The fetal heart beating rate was monitored, and blood samples of the fetus were taken to do the blood gas analysis and to detect the concentration of inflammatory cytokines (IL1, IL6, IL8, and TNFα). Results Two of the eight fetal lambs which was died in the operation because of pericardial tapenade. In the other six survived fetus, the PH was lower than after the surgery, and the concentrations of lactic acid, IL1, IL6, and IL8 are higher than after the surgery. There was no significant difference of PCO2,PO2 and TNFα between before and after the surgery. Conclusion The intrauterine cardiac intervention surgery can make the PH of fetal plasma lower and the concentrations of lactic acid and IL1, IL6, IL8 higher.
Objective To investgate the expression of p38 mitogen-activated protein kinase （p38MAPK） in lung tissue of rats with severe acute pancreatitis （SAP）， and to explore the relationship between p38MAPK and pulmonary capillary barrier injury. Methods Forty male and healthy Sprague-Dawley （SD） rats were randomly （random number method） divided into sham operation （SO） group and SAP group， then rats of SAP group were sub-divided into 3， 6， 12， and 24 h group， each group enrolled 8 rats， respectively. SAP model rats were established by injecting 5% sodium taurocholate solution retrograde into the biliopancreatic duct. ELISA method was used to test the serum tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）， and pathological changes in lung and pancreas tissues were observed by HE staining. Immunohischemistry method was used to detect phosphorylated p38 （p-p38） protein and aquaporin 1 （AQP1） protein of lung tissues. The expression level of AQP1 mRNA was measured by quantitative real-time PCR. Results Hyperemia， edema， and inflammatory cell infiltration were observed in lung tissues， abundance of necrosis， part gland structure fuzzy or even disappear were observed in pancreas tissues of all 4 time point groups. Compared with SO group， levels of serum TNF-α and IL-1β were significantly higher in 4 time point groups （P＜0.05）. Lower expression level of p-p38 protein was detected in lung tissues of SO group， while in the early stage of SAP （SAP 3 h group）， the expression level of p-p38 protein significantly increased， which peaked in 6 h group and was still higher than SO group in 24 h group （P＜0.05）. Compared with SO group， the expression levels of AQP1 mRNA and protein were significantly lower in all 4 time point groups （P＜0.05）， which had negative correlation with the levels of serum TNF-α，IL-1β， and the expression level of p-p38 protein （r=-0.87， P＜0.05；r=-0.88， P＜0.05；r=-0.78， P＜0.05）. Conclusion The decrease of AQP1 protein in lung tissue is one of the vital causes for pulmonary capillary barrier injury in SAP， which probably works by the activation of p38MAPK and the excessive release of inflammatory cytokines.
Objective To investigate the effects of glutathione S-transferase M5 (GSTM5) on the inflammation in human bronchial epithelial 16HBE cells and its possible molecular mechanisms. Methods Acute lung injury cell model was constructed with 16HBE cells induced by tumour necrosis factorα (TNF-α, 10 ng/mL). The cells were devided into a control group, a TNF-α group (TNF-α), a GSTM5 group (GSTM5+TNF-α), a negative control group (negative control plasmid+TNF-α). GSTM5-GFP plasmid and negative control plasmid were respectively transfected to the cells of the GSTM5 group and the negative control group using Lipofectamine2000. The contents of interleukin-6(IL-6), IL-8, IL-10 in the cell supernatant were measured by ELISA.The expression of nuclear factor-κB (NF-κB) mRNA was detected by RT-PCR, and the expression of NF-κB, phospho-NF-κB, p38, phospho-p38 protein were detected by Western blot. Results The GSTM5-GFP eukaryotic expression vector was successfully constructed and transfected successfully confirmed by fluorescence microscope. The contents of IL-6, IL-8, IL-10 in the TNF-α-induced cell supernatant were significantly higher than those in the control group(P < 0.05), and the contents of IL-6, IL-8, IL-10 in the GSTM5 group were lower than those in the TNF-α group (P < 0.05)with statistically significant difference. At the same time, the total NF-κB mRNA, phospho -NF-κB and phospho-p38 protein were increased in TNF-α stimulated cells compared with the control group (P < 0.05), while the GSTM5 group was lower than that in the TNF-α group and the negative control group (P < 0.05). Conclusion Overexpression of GSTM5 inhibits the phosphorylation of p38MAPK and NF-κB and down-regulates the inflammation of TNF-α-induced human bronchial epithelial 16HBE cells.
Myocardial remodeling is a common pathological physiology change for a variety of heart diseases under stimulation such as stress or ischemia. The engine body will release a lot of cytokines to promote the change of myocardial structure and ultimately lead to heart failure. Myocardial remodeling includes myocardial cells remodeling and the extracellular matrix remodeling. In recent years, we find that the function of adipose tissue is not only about energy storage, buffering to protect, supporting and filling, but also has a powerful function of secretion. Adipose tissue can secrete various adipocytokines, such as leptin, adiponectin, visfatin, omentin, angiotensin Ⅱ, and so on. Current studies have shown that adipocytokines and myocardial remodeling are intimated. And this article will summarize the function of adipocytokines on myocardial remodeling.
ObjectiveTo investigate the effect of etomidate and propofol on inflammatory cytokines and cortisol for patients with lung adenocarcinoma. MethodSixty patients scheduled for lung cancer surgery under general anesthesia were studied. All patients were randomly divided into an etomidate total intravenous anesthesia group (group E, 30 patients, 16 males and 14 females at age of 58.0±5.0 years) and a propofol total intravenous anesthesia group (group P, 30 patients, 17 males and 13 females at age of 55.0±5.0 years), with 30 patients in each group. ResultsThe concentration of IL-6 in serum of patients in the two groups at time points T1, T2 and T3 was significantly higher than those at time point T0 (P < 0.01). The concentration of IL-10 and TNF-α in serum of patients at time points T1 and T2 was significantly higher than those at time point T0 (P < 0.01). And the difference of the concentration of TNF-α in serum of patients at time points T0 and T3 was not statistically significant (P > 0.05). The level of Cor of patients in the group E at time point T0 was slightly higher than those at time point T1, but lower than that at time points T2 and T3. There was no statistical difference in the concentration of IL-6 and TNF-α in serum of patients between the two groups. The level of IL-10 of patients in the group E at time points T2 and T3 was lower than those in the group P (P < 0.05), but no significant difference was observed at the other time points. The concentration of Cor in the patients in the group E at time point T1 was lower than that in the group P (P < 0.01), but no significant difference was observed either at the other time points. ConclusionThe effect of etomidate used for maintenance of general anesthesia on the inflammatory factors is essentially similar to that of propofol.
Objective To investigate the protective effect of castanospermine (CS) on renal injury induced by severe acute pancreatitis (SAP) in rats and its possible mechanism. Methods Twenty-four SPF adult male Sprague Dawley rats were randomly divided into three groups: shame operation group (SO group, n=8), SAP group (n=8), and CS group (n=8). SAP models were induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) in biliopancreatic duct in the SAP group and the CS group. CS solution (200 mg/kg) was immediately administered via intraperitoneal injection after the induction of pancreatitis in the CS group. Rats in the SO group were subjected to a sham surgery that the pancreas and duodenum were flipped a number of times. All rats were sacrificed at 12 h after modeling. Blood samples were collected by inferior vena cava puncture, and serum activities of amylase (AMY), levels of blood urea nitrogen (BUN) and creatinine (Cr) were measured by using a fully automatic chemistry analyzer. The head of pancreas and renal tissues were harvested and pathological change was observed under the light microscope. Expressions of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), intercellular adhesion molecule-1 (ICAM-1), and Caspase-3 in renal tissues were evaluated by immunohistochemistry assay. Results ① Compared with the SO group, the damages of the pancreas and kidney tissues were significantly worse in the SAP group, and the above damages in the CS group were significantly decreased when comparing with the SAP group. ② Compared with the SO group, the serum activities of AMY, levels of BUN and Cr were significantly increased in the SAP group (P<0.05). The serum activities of AMY, levels of BUN and Cr in the CS group were significantly lower than those of the SAP group (P<0.05). ③ Compared with the SO group, the integrated optical density (IOD) of NF-κB, TNF-α, ICAM-1, and Caspase-3 in renal tissues were significantly increased in the SAP group (P<0.05), and the above indicators in kidney tissues of the CS group were significantly decreased when comparing with the SAP group (P<0.05). Conclusions CS can mitigate severe acute pancreatitis-induced renal injuries in rats, it ameliorates renal injury and improves renal function. The mechanism for the above improvements is that CS can widely inhibit the activation of NF-κB, and then downregulate the expressions of TNF-α, ICAM-1, and Caspase-3.