Apoptosis in heart of multiple organ dysfunction syndrome rats at high altitude

AIM:To study apoptosis in heart of multiple organ dysfunction syndrome (MODS) rat at high altitude. METHODS:The two-hit model of MODS rat was used at two different altitude(1510m, 3900m). Hemorrhage was induced in Wistar rats by catheterizing the femoral artery until a mean arterial pressure was 35 mmHg and maintained for 1 hour. Rususcitation was performed with lactated Ringer's solution at 24 h after hemorrhage, cecal ligation and puncture(CLP) was performed.Then rats were killed at 3 h, 6 h, 12 h and 24 h after CLP, and myocardium sample was excised and stored in liquid nitrogen. Apoptosis in heart was determined by DNA agarose gel electrophoresis, flow cytometry(FCM), transmission electron microscope(TEM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL). RESULTS:The special ladder pattern for apoptosis was seen in myocardium sample at high altitude(3 900 m) group.The apoptotic rate in myocardium was higher in high altitude(3 900 m) group than that in lower altitude(1 510 m) group(P<0.01).TUNEL showed that there were positive cells in myocardium in high altitude(3 900 m) group. Apoptotic body was also seen in myocardium in high altitude(3 900 m) group by TEM. CONCLUSION:Apoptosis was induced in myocardium of MODS rats at high altitude, which might play a role in heart failure. The environment of high altitude accelerates the impairment of myocardium in MODS rats.     

Ultrastructural,physiological and biochemical study on multiple organ dysfunction syndrome in the elderly rats

AIM:By studying the ultrastructure of organs (hearts, brains, lungs, livers, kidneys) and vascular endothelial cells, and changes of physiological and biochemical indexes with multiple organ dysfunction syndrome in the elderly rats, their signification and their developing rules were analyzed.METHODS:Wistar rats(Half of Wister rats were three-month-old and another half of twenty-month-old) were anaesthetized and dealt with cecal ligation puncture (CLP) operation. They were defined as MODSE group and MODSY group, respectively. They were perfused through all vascular circulation by 1% glutaric dialdehyde. These slices were observed under transmission electron microscopy. RESULTS:Physiological and biochemical indexes at 24 huor in elder rats were shown significant difference compared with the younger rats (P＜0.05). Moreover, Ultrastructures of all organs and vascular endothelial cells at 24 h after operation in MODSE group have shown to be more serious injury than that in MODSY group.CONCLUSIONS:According to ultrastructure observations, the injuries of organ and vascular endothelial cells with MODSE or MODSY occurred. Moreover, these damages in MODSE group occurred more seriously. The injuries of organ and vascular endothelial cells with MODSE or MODSY were accorded with physiological and biochemical changes.     

The possible nitric oxide mechanism of Rhodiola on the multiple organ dysfunction syndrome after burns

AIM:To study the effects of Rhodiola(Rho), nitric oxide (NO), hemoglobin (HB) on the multiple organ dysfunction syndrome(MODS) in early stage after burn in rabbits. METHODS:The rabbits were divided into the sham burn group(SB), burn group (B), orally taken Rho group (R), burn and Rho therapy group (BR). The changes of hemodynamics were monitored. The index of pulmonary permeability was calculated. These data reflected separately the functions of heart, liver, lung, kidney and blood coagulation system were also determined. NO contents in serum and bronchoalveolar lavage fluid (BALF) were measured by Griess method. The levels of serum HB were measured. RESULTS:① The dysfunctions of heart, liver and kidney achieved the criterion of MODS in group B at 48 h postburns (B^{48 h}). The NO content of group B significantly increased in serum and BALF at B^{48 h}. ② The cardiac index (CI) and creatine phosphokinase (CK), urea nitrogen (BUN) markedly raised or decreased in group BR at 48 h postburns (BR^{48 h}) than B^{48 h}. The NO contents in serum and BALF markedly raised. ③ HB contents in serum markedly raised in group B and BR at 0 h postburns (B^{0 h}, BR^{0 h}) than group SB, B^{48 h}, R, BR^{48 h}, but NO was reverse. CONCLUTIONS:① HB contents in serum markedly raised at 0 h postburns, but NO was reverse. ② Rho promoted the increases of NO synthesis and the blood perfusion of organs, which might be one of mechanisms to prevent the development of MODS.     

Effect of Siduqing decoction,a Chinese medicine,on survival rate and multiple organ dysfunction in mice challenged with LPS

AIM: To investigate the effect of siduqing decoction, a Chinese medicine, on survival rate and multiple organ dysfunction in mice challenged with LPS. METHODS: Mice were administered intragastrically with Siduqing decoction or distilled water (0.2 ml/10 g) twice a day for 3 days, two hours after Chinese herbal medicine treatment on day 3, LPS or normal saline was injected intraperitoneally, and survival rates in each group were recorded at 12-h intervals. In another experiment, mice were sacrificed at 12 h after LPS, lung, liver, kidney and small intestine were collected and processed for the H & E staining. In addition, Blood was collected at 10 h after LPS injection for determining alanine aminotransferase (ALT) activity, blood urea nitrogen (BUN) and creatinine (Cr) contents. RESULTS: At 96 h after LPS injection, the survival rate (27%, n=34) was lower in LPS group than Siduqing treatment group (65%, n=31, P<0.05). ALT activity, BUN and Cr contents in serum were higher in LPS group than control group, Siduqing treatment significantly attenuated a increase in ALT activity, BUN and Cr content in serum induced by LPS. Histological examination showed inflammatory injury in the lung and intestine, hemorrhage in the lung and kidney, degeneration, necrosis in the liver and kidney, while Siduqing treatment attenuated pathological changes induced by LPS. CONCLUSION: These data indicate that Siduqing has a protective effect against LPS-induced multiple organ injury and increases survival rate in mice challenged with LPS.     

Effects of oxidized low density lipoprotein and vitamin E on the levels of IL-6,IL-8 and TNF-α in cultured human umbilical vein endothelial cells

AIM:To investigate effects of OX-LDL and VitE on the levels of IL-6,IL-8 and TNF-α in human umbilical vein endothelial cells(HUVEC).METHODS: Human umbilical vein endothelial cells were obtained by in vitro culture. HUVEC treated with or without Vit E was incubated with OX-LDL, and the levels of IL-6, IL-8 and TNF-α were determined by enzyme-linked immunosorbent assy technique. RESULTS:50 μg/L,100 μg/L, 200 μg/L OX-LDL induced the release of IL-6,IL-8 and TNF-α by HUVEC in a dose-dependent manner. Compared with the control group , the levels of IL-6 and IL-8 were significantly increased at 6-12 h of stimulation with OX-LDL . Maximal levels of IL-6 and IL-8 occurred after 24-36 h, reaching a plateau maintained for at least 48 h. TNF-α rose after 2-6 h in HUVEC, and reached a maximum after 12 h. In contrast to IL-6 and IL-8, TNF-α declined after 48 h. However, when VitE (50 mg/L,100 mg/L,200 mg/L)was added, it can significant inhibited the release of IL-6, IL-8 and TNF-α in a dose-dependent manner, and after 48 h these cytokines have no diference between OX-LDL+VitE groups and OX-LDL groups. CONCLUSION: OX-LDL can obviously stimulate the production of IL-6,IL-8 and TNF-α in vascular endothelial cells, which can significantly be inhibited by VitE in a short time.     

Effect of normal mesenteric lymph on multiple organ injury in mice with endotoxic shock

AIM: To observe the effects of normal mesenteric lymph (NML) on the lung, heart and liver injuries and the phosphorylation levels of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK) in the mice with endotoxic shock (ES). METHODS: The NML was drained form health male BALB/c mice for the intervention of ES after the removal of cellular constituent. Lipopolysaccharide (LPS, 35 mg/kg) was intraperitoneally injected into the mice for the establishment of ES model. After 60 min of LPS injection, the administration of NML (1/15 of whole blood volume) was performed through the femoral artery in NML+ES group. Meanwhile, the mean arterial pressure (MAP) was monitored during the experiment. At 6 h after intraperitoneal injection of LPS or the corresponding time point, blood samples were harvested from the heart through apical centesis for determination of the biochemical indexes to reflect myocardial and hepatocyte injuries. Simultaneously, the lung, heart and liver tissue specimens from a fixed location were harvested for the observation of histomorphology and the measurement of phosphorylation levels of p38 MAPK, ERK1/2 and JNK. RESULTS: Compared with sham shock (SS) group, MAP in ES group and NML+ES group remarkably decreased at multiple time points after intraperitoneal injection of LPS. However, MAP in NML+ES group at 80 min, 90 min, 190 min, 210 min, 240 min, 250 min, 340 min, 350 min, and 360 min were significantly increased compared with ES group. There were normal structures in the lung, liver and myocardium of the mice in SS group, while the morphological damages of these tissues appeared in ES group. Meanwhile, the damages were attenuated in the mice of NML+ES group. The activities of AST, ALT and CK-MB in the plasma in ES group were remarkably higher than those in SS group. The CK-MB activity in NML+ES group was also increased compared with SS group, and the activities of AST and LDH-1 were lower than those in ES group. At 6 h after LPS injection, the phosphorylation levels of p38 MAPK, ERK1/2 and JNK in the lung tissues were remarkably increased. Meanwhile, no statistical difference of these indexes between the myocardial and hepatic tissues was observed. NML intervention decreased the phosphorylation levels of p38 MAPK in the lung tissues, and p38 MAPK, ERK1/2 and JNK in the myocardial tissues. CONCLUSION: The NML administration alleviates multi-organ injuries and reduces the phosphorylation level of p38 MAPK in the lung tissues in the mice subjected to ES.     

Expression of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 and collagen type IV in lung of rats with multiple organ dysfunction syndrome

AIM: To determine the expression of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of me-talloproteinase-1 (TIMP-1) and collagen type IV (IV-C) in the lung of rats with multiple organ dysfunction syndrome (MODS) and to investigate the mechanism of lung injury in MODS. METHODS: Adult male Sprague-Dawley (SD) rats (n=40) were randomly divided into sham control group and cecal ligation and puncture (CLP) model group. The rats in CLP group were divided into 4 subgroups as different intervals (6 h, 12 h, 24 h and 48 h), and there were 8 rats in each group. The rat model of MODS was established by CLP. All rats were sacrificed at various intervals. The functions of the liver, kidney and lung were determined by blood biochemical and blood gas analysis. The morphological changes of the lung tissues were observed with HE staining. The serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, MMP-9 and TIMP-1 were measured by ELISA. The expression of MMP-9 and TIMP-1 in the lung tissues was detected by RT-PCR and immunohistochemistry, and the expression of IV-C in the lung tissues was detected by immunofluorescence and Western blot. RESULTS: Compared with sham control group, the functions of the liver, kidney and lung were damaged at different degrees in model groups. No histopathological change in the lung tissues of sham control group was found, and the lung injury was serious in model groups. Compared with sham control group, the serum levels of TNF-α, IL-1β, MMP-9 and TIMP-1 in model groups increased significantly (P<0.05) and peaked at the interval of 12~24 h after modeling (P<0.01). The expression of MMP-9 and TIMP-1 in the lung tissues of model groups increased, and peaked at 12 and 24 h, respectively (P<0.01). The protein level of IV-C in MODS 6 h group was not changed as compared with control group, while that at the interval of 12~48 h after modeling was significantly decreased and dropped to the lowest at 24 h (P<0.01). CONCLUSION: MMP-9 and TIMP-1 play important roles in lung injury of MODS rats by regulating the synthesis and decomposition of IV-C which is the main component of extracellular matrix.     

Effects of atorvastatin on expression of PAPP-A induced by TNF-α and IL-1β in rat aortic endothelial cells

AIM: To investigate the effects of atorvastatin on the expression of pregnancy-associated plasma protein A(PAPP-A)induced by TNF-α and IL-1β in endothelial cells. METHODS: The rat aortic endothelial cells were isolated from thoracic aortas and cultured by the tissue explant method. The cells in passage 3-4 were used in the experiment and were randomly divided into 4 groups: blank control group: the cells were treated without any intervention; atorvastatin concentration groups: the cells were incubated with atorvastatin at the concentrations of 0.1, 1 and 10 μmol/L for 24 h; atorvastatin time groups: the cells were incubated with atorvastatin at the concentration of 10 μmol/L for 6 h,12 h and 24 h; atorvastatin+inflammatory factors groups: the cells were pre-incubated with 60 μg/L TNF-α or 20 μg/L IL-1β for 1 h, then different concentrations of atorvastatin (0.1, 1.0, 10 μmol/L) were added for 6 h,12 h and 24 h. MTT reduction assay was used to observe the cell proliferation. The mRNA expression of PAPP-A was detected by RT-PCR. The protein level of PAPP-A in the supernatants of cultured cells was measured by ELISA. RESULTS: Compared with blank control group, no significant change of cell proliferation was observed after the intervention of atorvastatin and TNF-α/IL-1β for 3 h, 6 h, 12 h, 24 h and 48 h, indicating that the drugs had no toxic effects on the cells. No significant difference of PAPP-A expression between atorvastatin groups and blank control groups was found. Compared with TNF-α groups and IL-1β groups, PAPP-A expressions in atorvastatin intervention groups significantly decreased. The protein level of PAPP-A was gradually decreased with the raised concentration of atorvastatin and the prolonged time in a concentration- and time-dependent manner. CONCLUSION: Atorvastatin doesn't influence the PAPP-A expression, but inhibits the expression of PAPP-A activated by inflammatory factors in a concentration- and time-dependent manner in primary cultured rat aortic endothelial cells.     

Changes of IL-4, IL-10, IL-12 levels in rat serum and lung during acute respiratory distress syndrome induced by oleic acid

AIM:To observe the dynamic changes of interleukin-4(IL-4), IL-10, IL-12 in rat serum and lung tissues during acute respiratory distress syndrome (ARDS).METHODS:The ARDS model of rats was induced by intravenous injection of oleic acid. The levels of IL-4, IL-10, IL-12 in serum and the supernatant of lung tissues were measured by enzyme linked immunosorbent assay (ELISA).RESULTS:The Levels of serum and lung IL-10, IL-12 in ARDS rats were increased in 4 h, 8 h, 16 h group compared with control group. The levels in IL-10 in serum in 16 h group and IL-10 in lung tissues of 8 h group were lower than that in 4 h group. The Levels of IL-4 in serum in 4 h, 8 h group were higher than that in control group, while IL-4 in 16 h group was lower than that in 8 h group. IL-4 of lung tissues in 4 h, 8 h, 16 h group were increased significantly, but in 16 h group were lower than that in 8 h group. The biggest changes of pulmonary coefficient and histopathology were observed at 4 h after injection of oleic acid.CONCLUSIONS:IL-4, IL-10 and IL-12 might play important roles in inflammatory reaction induced by oleic acid. The pro-and anti-inflammatory cytokines produced successively during ARDS.The relationship between unbalanced cytokines and lung injury in ARDS needs to be further studied.     

Observation of cardiomyocytes stimulated by TNF-α, IL-1β, LPS in vitro

AIM: To observe the changes of cardiomyocytes after stimulation by TNF-α, IL-1β, LPS.METHODS: Cardiac ventricular myocytes were cultured in vitro. Different doses of TNF-α, IL-1β, LPS were added to stimulate the cardiomyocytes, the hypertrophy of cardiomyocytes 8 h, 24 h, and 48 h after stimulation was determined and the apoptosis were also observed 24 h, 48 h, 72 h after stimulation. RESULTS: Compared to the normal myocytes, the cardiomyocytes were hypertrophied after stimulation by 10 μg/L, 15 μg/L of TNF-α, 20 μg/L, 100 μg/L of IL-1β and 10 mg/L, 15 mg/L, 20 mg/L of LPS, and the effect was dose-dependent, the strongest effect was showed in 24 h. Moreover, 20 μg/L of TNF-α, 100 μg/L of IL-1β and 30 mg/L of LPS caused cardiomyocyte apoptosis, especially in 72h. CONCLUSION: TNF-α, IL-1β, LPS induced the cardiomyocyte hypertrophy and apoptosis, suggesting the inflammation may be the main cause of cardiovascular disease.     

Alteration of serum IL-6, IL-8, TNF-α and sICAM-1 in patients with pre-menstruation recurrent aphthous ulceration

AIM:To investigate the serum levels of interleeukin-6(IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha(TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1)in female patients with pre-menstruation recurrent aphthous ulceration(RAU).METHODS:Serum levels of IL-6, IL-8, TNF-α and sICAM-1 in 21 pre-menstruation RAU patients were examined using ELISA technique, and compared to 10 healthy individuals and 22 the female RAU patients unrelated to menstrual cycle.RESULTS:The serum levels of IL-6, IL-8, TNF-α in patients with pre-menstruation RAU were not only significantly higher than that in the normal control group(P＜0.01), but also higher than that in the RAU patients without pre-menstruous recurrence (P＜0.01).The level of serum TNF-α in the RAU patients without pre-menstruous recurrence was slightly higher than that in the normal control group(P＜0.05).The level of sICAM-1 had not changed.CONCLUSION:The serum levels of IL-6, IL-8 and TNF-α increase in pre-menstruation RAU patients, which might play a role in local lesion of RAU.     

Effects of Huaiyu decoctum on serum concentrations of TNF-α, IL-6 and IL-10 in rats after anorectal operation

ATM: To investigate the effects of Huaiyu decoctum on the serum concentrations of TNF-α, IL-6 and IL-10 in rats after anorectal operation. METHODS: Sprague-Dawley rats (n=40) were randomly divided into 4 groups:normal group, model group, low-dose Huaiyu decoctum group and high-dose Huaiyu decoctum group. The concentrations of TNF-α, IL-6 and IL-10 in the rat serum were measured by ELISA. The pathologic changes of the anorectal tissues were observed under microscope with HE staining. The protein expression of ICAM-1, VCAM-1 and NF-κB was determined by Western blotting. RESULTS: After Huaiyu decoctum administration, TNF-α and IL-6 concentrations in the serum were significantly decreased, and IL-10 concentration was increased as compared with model group. Moreover, Huaiyu decoctum markedly attenuated edema and hyperemia in the rats after anorectal operation. The protein expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and NF-κB in the anorectal tissues was obviously inhibited by Huaiyu decoctum treatment. CONCLUSION: Huaiyu decoctum improves the recovery of anorectal tissues after operation by decreasing the serum concentrations of TNF-α, IL-6 and IL-10, and reducing the protein expression of ICAM-1, VCAM-1 and NF-κB in the anorectal tissues.     

Effect of recombinant rat CC16 protein on LPS-induced expression of TNF-α, IL-6 and IL-8 in rat tracheal epithelial cells

AIM: To explore the effect of recombinamt rat CC16 protein (rCC16) on LPS-induced expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 in the rat tracheal epithelial (RTE) cells.METHODS: The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS (0.1 mg/L) treatment for further 24 h. The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR. The cell culture supernatants were collected for analyzing the protein levels of TNF-α, IL-6 and IL-8 by ELISA. In addition, the nuclear translocation of nuclear factor-κB (NF-κB) p65 was tested by Western blot.RESULTS: rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent (0~2 mg/L) manner, as demonstrated by RT-qPCR and ELISA. However, no concentration-dependent manner between the dose of rCC16 and TNF-α expression was observed, and rCC16 inhibited LPS-induced TNF-α expression at lower concentration (0.5 mg/L). rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION: rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 production through inactivation of NF-κB activity in RTE cells.[KEY WORDS] CC16 protein; Airway inflammation; LPS; Inflammatory mediators; Nuclear factor-κB     

Expression of IL-1α and TNF-α modified by Toll-like receptor 4 genetic polymorphism is associated with colorectal carcinoma

AIM: To investigate the association of D299G, T399I and A896G polymorphisms of Toll-like receptor 4 (TLR4) and colorectal carcinoma (CRC). METHODS:
The genotypes of these 3 loci among 268 patients with CRC and 268 healthy controls were determined by polymerase chain reaction-restriction fragment lengthy polymorphism (PCR-RFLP). The protein levels of IL-1α, IL-8, TGF-β and TNF-α in the homogenate of CRC biopsies were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: No significant difference of the genotype frequencies of TLR4 A896G and D299G between the cases and the controls was observed. CT combined TT genotype of T399I was significantly associated with increased CRC risk. The individuals with the T allele of T399I showed a 1.843-fold increase in CRC risk as compared with the C allele. The concentrations of IL-1α and TNF-α in CRC biopsies were significantly elevated in the individuals with the genotype of T399I CT combined with TT as compared with the genotype of CC. CONCLUSION: TLR4 T399I promotes the development of CRC by modifying the expression of IL-1α and TNF-α in CRC tissues.     

Relationship between levels of IL-2, TNF-α and nitric oxide in aqueous humor after intraocular lens implantation

AIM: To investigate the relationship between the level of interleukin-2 (IL-2), tumour necrosis factor-α (TNF-α) and nitric oxide (NO) in aqueous humor after intraocular lens implantation. METHODS: New Zealand rabbits were divided randomly into three groups: (1) control group; (2) extracapsular cataract extraction group (ECCE); (3) extracapsular cataract extraction and posterior chamber intraocular lens implantation group (ECCE+IOL). The inflammation in all experimental rabbit eyes was observed via zoom-photo slit-lamp microscope on 1, 3, 7, 14 d and 30 d postoperation. Meanwhile, aqueous humor was drawn for white blood cell (WBC) counting and classifying and for determining IL-2, TNF-α and NO₂^-/NO₃^- contents. RESULTS: (1) The level of IL-2 and TNF-α and NO₂^-/ NO₃^- in aqueous humor of ECCE+IOL group were higher than that in ECCE and control at 1 to 14 days postoperation, respectively, it increased to peak value at 3 to 7 days postoperation and decreased gradually two weeks postoperation; (2) The changes in IL-2, TNF-α and NO₂^-/NO₃^- in each group were basically similer; (3) The changes of IL-2 and TNF-α level were closely related with NO content in aqueous humor (r=0.69, P<0.01 and r=0.98, P<0.01). CONCLUSION: IL-2, TNF-α and NO play an important role in intraocular inflammation intraocular lens implantation.     

Effect of NOD8 on production of nitric oxide,IL-1β and TNF-α in LPS-treated macrophages

AIM: To investigate the effect of NOD8 on lipopolysaccharide (LPS)-induced releases of nitric oxide (NO), tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells. METHODS: The plasmids of pEGFP-C2 and pEGFP-NOD8 were transfected into RAW264.7 cells respectively. The transfected and non-transfected cells were stimulated by LPS for 0, 6, 12 and 24 h. NO production was evaluated by Griess reagent assay, and the levels of IL-1β and TNF-α were measured by ELISA. The protein expression of NOD8 and the nuclear translocation of nuclear factor κB (NF-κB) p65 subunit were detected by Western blotting. The level of activated caspase-1 was determined by fluorimetric method. RESULTS: Compared with pEGFP-C2 group, the protein expression of NOD8 was significantly elevated in pEGFP-NOD8+LPS group. The releases of NO, IL-1β and TNF-α were obviously increased after RAW264.7 cells were treated with LPS for 6 h, 12 h and 24 h, and while the secretion of NO was significantly reduced in the cells transfected with pEGFP-NOD8 and induced by LPS for 12 h and 24 h, and the release of IL-1β was also significantly reduced at 6 h, 12 h and 24 h. However, no significant difference of TNF-α release was observed between pEGFP-C2+LPS group and pEGFP-NOD8+LPS group. The activation of caspase-1 in RAW264.7 cells stimulated with LPS for 6 h, 12 h and 24 h was markedly increased, and the expression of NF-κB p65 subunit in the cytoplasm was significantly decreased, indicating that p65 nuclear translocation was increased. In addition, the activation of caspase-1 and the nuclear translocation of p65 were significantly inhibited in pEGFP-NOD8+LPS group. CONCLUSION: NOD8 suppresses the releases of LPS-induced NO and IL-1β in RAW264.7 cells by inhibiting the activation of caspase-1 and NF-κB.     

Effects of TNFα and IL-10 on cytomegalovirus infection in human embryonic lung fibroblasts

AIM:To study effects of tumour necrosis factor alpha(TNFα) and interleukin-10(IL-10) on human cytomegalovirus AD169 (HCMV AD169) infection in human embryonic lung fibroblasts (HEL),and the ability of the infected HEL to produce TNFα. We have attempted to understand the effect of cytokines in the immune of HCMV infection. METHODS:TNFα、IL-10 were added separately with different concentrations before 24 h HCMV infection of HEL to study the effect of TNFα and IL-10 on multiplication of HCMV. HCMV was incubated with HEL, TNFα in culture supernatants were measured at 4, 24, 48, 72, 96 h postinfection. The level of TNFα was measured by enzyme-linked immunosorbent assay( ELISA). RESULTS:The addition of TNFα with the concentration of 10-100 μg/L or IL-10 with the concentration of 1-10 μg/L before 24 h HCMV infection of HEL could remarkably inhibited the multiplication of HCMV in HEL. Level of TNFα didn't increase in infected cell supernatants. CONCLUSION:TNFα and IL-10 play an important role in the immune of HCMV infection.     

Effects of IL-6 and IL-1α on the peripheral blood polymorphonuclear neutrophil apoptosis postburn in rats

AIM:To study the effects of IL-6 and IL-1α on the blood polymorphonuclear-neutrophils(PMN) apoptosis postburn.METHODS:Wistar rats inflicted by 30% total body surface area (TBSA) Ⅲ degree scalding were employed as the model. PMN were isolated by density gradient centrifugation using Percoll-hypaque and labeled with TdT-mediated and dUTP nick end labeling (TUNEL) and analyzed by flow cytometric analysis. The intracellular caspase-3 activation and the serum levels of IL-6 and IL-1α were analyzed by fluorometric immunosorbent enzyme assay and enzyme-linked immunosorbent assay, respectively.RESULTS:The serum IL-6 levels (μg/L) in groups of 3, 6, 12, 24 and 48 h postburn (9.14±1.16, 12.49±1.14, 3.01±0.75, 1.41±0.28 and 1.56±0.43 in turn) and IL-1α (ng/L) in groups of 3, 6, 12 h postburn (90.08±8.39, 320.93±14.48 and 47.84±5.19) were much higher than IL-6 (0.24±0.07) and IL-1α (27.65±4.86) in control group (P＜0.05), respectively. The relative proportions of apoptotic PMN(%) in groups postburn were 9.89±2.00, 4.98±1.35, 1.31±0.72, 2.49±1.87 and 6.88±1.13 in turn, which were obviously less than 13.66 ± 3.88 in the control group. The results of intracellular caspase-3 activation in PMN were observed to be consistent with the results of apoptotic PMN analysis in experimental groups.CONCLUSION:IL-6 and IL-1α are the important factors to induce PMN apoptosis delay that occurred obviously postburn. And decrease in the activity of intracellular caspase-3 of PMN may be involved in their mechanisms.