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.     

Effects of immunoglobulin on circulating proinflammatory cytokines in children with hand-foot-mouth disease

AIM：To study the mechanism of intravenous administration of immunoglobulin for treatment of hand-foot-mouth disease. METHODS：Sixty-four children with hand-foot-mouth disease were randomly divided into treatment group and control group. The children in control group (n=32) were treated with ribavirin and given normal supportive therapy. The children in treatment group (n=32) were given ribavirin and normal supportive therapy plus intravenous administration of immunoglobulin (500 mg·kg-1·d-1) for 3 d. The levels of TNF-α, C-reactive protein (CRP) and IL-6 in venous blood were observed. RESULTS：The levels of TNF-α, CRP and IL-6 in treatment group and control group were all higher than those in the normal children. The levels of TNF-α, CRP and IL-6 in treatment group were similar to those in control group before treatment. The levels of TNF-α, CRP and IL-6 in treatment group were significantly lower than those in control group after treatment with immunoglobulin for 3 d. CONCLUSION：The systemic inflammatory response syndrome is one of the possible mechanisms of hand-foot-mouth disease. Intravenous administration of immunoglobulin decreases the proinflammatory cytokines in the patients with hand-foot-mouth disease.     

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.     

Effect of sedation by infusion of dexmedetomidine on inflammatory factors in patients with severe trauma

AIM: To study the effect of short-term sedation by infusion of dexmedetomidine (DEX) on the inflammatory factors in the patients with severe trauma. METHODS: Sixty patients with severe trauma in ICU were randomly divided into 3 groups: DEX group (n=20), midazolam (MDZ) group (n=20) and control group (without any sedatives, n=20). Dexmedetomidine at an initial loading dose of 1~2 μg/kg was administered intravenously over 10 min to the patients in DEX group before the sedation procedure of continuous infusion at dose of 0.2~0.7 μg·kg^-1·h^-1 by a 50-mL infusion syringe was perform. Midazolam at dose of 0.03~0.3 mg/kg was also administered intravenously immediately before the procedure of a continuous infusion at dose of 0.03~0.2 mg·kg^-1·h^-1. The doses of DEX or MDZ for the patients receiving short-term (48 h) sedation were adjusted according to the Ramsay sedation scale scores. The serum levels of interleukins (IL-1 and IL-6), tumor necrosis factor α (TNF-α) and C-reactive protein (CRP) were measured at the time points of the baseline (before the start of the study), 24 h and 48 h of sedative infusion. RESULTS: The levels of TNF-α, IL-1, IL-6 and CRP in the 3 groups of severe traumatic patients at the time when they were admitted to the hospital were significantly higher than those of the normal values. The levels of TNF-α, IL-1, IL-6 and CRP in control group gradually increased at 24 h and 48 h, while those in DEX group and MDZ group significantly declined. No significant difference of TNF-α, IL-1, IL-6 and CRP levels between DEX group and MDZ group was observed at 24 h, but the changes were significantly different at 48 h. CONCLUSION: Dexmedetomidine reduces over-stress responses to a certain extent in the traumatic patients to prevent the further production and release of inflammatory mediators, thus contributing to the stability and recovery of the patients with severe trauma.     

Serum acute phase reactants sCRP,SAA, IL-6 in acute coronary syndrome and its clinical significance

AIM: To investigate the possible involvement of CRP, SAA, IL-6 and its significance in acute coronary syndrome. METHODS: Serum SAA, IL-6 and sCRP levels in 40 patients with AMI, 40 patients with UAP and 40 controls were measured by means of ELISA. RESULTS: Serum SAA, IL-6 and CRP were significantly higher in ACS group (AMI or UAP) than in control group. In the Spearman correlation analysis , we found a correlation among IL－6, SAA, sCRP, fibrinogen both in the AMI group and UAP group. There was no correlation between CK-MB and serum SAA, IL-6 and sCRP levels either in AMI group or in UAP group. In the AMI group, there was no difference of serum SAA,IL-6 and sCRP levels when divided into ≤6 score group and >6 score group using Wagner's QRS scoring system. CONCLUSIONS: Serum levels of certain inflammatory markers have some diagnostic value for ACS. The high levels of serum SAA, IL-6 and sCRP are not mainly caused by myocardial death, but by the inflammation in the multifocal unstable plague.     

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.     

Bone marrow-derived mesenchymal stem cell-conditioned medium atte-nuates LPS-induced acute lung injury in mice

AIM:To explore the effects of bone marrow-derived mesenchymal stem cells-conditioned medium (MSCs CdM) on lipopolysaccharide (LPS)-induced acute lung injury. METHODS:Lung injury was induced in mice by intraperitoneal injection of LPS. The mice were given a tail vein injection of MSCs CdM or normal saline 1 h after LPS administration. The mice were killed by an intraperitoneal injection of pentobarbital 6 h after LPS injection for either bronchoalveolar lavage fluid (BALF) and serum collection or lung histological analysis. RESULTS:Compared with control group, the BALF levels of protein, interleukin-10 (IL-10) and keratinocyte growth factor (KGF), the serum levels of tumor necrosis factor α (TNF-α) and IL-6, and the myeloperoxidase (MOP) activity in the lung tissues were significantly higher in LPS group, and severe pathological damages in the lung tissues were also observed. Treatment with MSCs CdM significantly reduced the BALF prtein level, the seum TNF-α and IL-6　levels and the lung MPO activity, and attenuated the lung pathological damages, but further increased the levels of IL-10 and KGF in the BALF. CONCLUSION:Treatment with MSCs CdM attenuates the lung injuries induced by LPS, which may be via regulating the expression of TNF-α, IL-6, IL-10 and KGF.     

Influence of 8-week swimming on peripheral neuropathy in type 2 diabetic rats

AIM: To explore the influence of long-term swimming on peripheral neuropathy in type 2 diabetic rats. METHODS: Male Wistar rats were fed with a high-fat and high-fructose diet, and injected with streptozocin to establish a model of type 2 diabetes mellitus. The rats were randomly divided into 4 groups: blank control group (C group), exercise control group (CE group), diabetes mellitus group (DM group) and diabetes mellitus+exercise group (DME group). The rats in CE group and DME group received 8-week swimming training (6 d/week). The training time was 20, 30 and 45 min in the first 3 d,respectively, and then it increased to 60 min a day. Eight weeks later, the motor nerve conduction velocity (MNCV) and the levels of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and C-reactive protein (CRP) in sciatic nerve tissues of the rats were measured. The morphological changes of the sciatic nerve were also observed under light microscope. RESULTS: Compared with DM group, 8-week swimming obviously accelerated the MNCV (P<0.05), decreased the levels of TNF-α, IL-6 and CRP in DME group (but no significant difference, P>0.05). The obvious nerve injury in DM group was observed. However, the pathological change of the sciatic nerve in DME group was relieved. CONCLUSION: Eight-week swimming training significantly accelerates the MNCV, attenuates the nerve injury in diabetic rats and has protective effect on peripheral nerve, which may be correlated with relieving the inflammatory reaction.     

Effects of rosiglitazone on expression of SOCS1/SOCS3 and inflammatory reaction in RAW 264.7 cell-derived foam cells

AIM: To investigate whether perioxisome proliferator-activated receptor γ (PPARγ) ligand rosiglitazone regulates suppressor of cytokine signaling 1 (SOCS1) and SOCS3 expression as well as pro-inflammatory/anti-inflammatory responses in RAW 264.7 cell-derived foam cells. METHODS: The concentrations of TNF-α, IL-6 and IL-10 in the cultured supernatant of RAW 264.7 cell-derived foam cells were detected by ELISA, and the ratios of TNF-α/IL-10 and IL-6/IL-10 were calculated. RT-PCR and Western blotting were used to analyze the effects of rosiglitazone on the expression of SOCS1 and SOCS3 at mRNA and protein levels. RESULTS: The concentrations of TNF-α, IL-6 and IL-10, and ratios of TNF-α/IL-10 and IL-6/IL-10 in foam cell group were obviously higher than those in control group, but the concentrations of the above factors in oxidized low-density lipoprotein (ox-LDL) +rosiglitazone group were apparently lower than those in foam cell group. The expression of SOCS1 and SOCS3 at mRNA and protein levels in oxLDL+rosiglitazone group was apparently higher than that in control and foam cell group. CONCLUSION: PPARγ ligand rosiglitazone up-regulates the expression of SOCS1 and SOCS3 at mRNA and protein levels and regulates the balance of pro-inflammatory/anti-inflammatory responses in RAW 264.7 cell-derived foam cells.     

Effect of granulocyte colony-stimulating-factor on acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation in a murine model

AIM: To explore the impact of granulocyte colony-stimulating factor (G-CSF) on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in a murine model and its possible mechanisms. METHODS: Male C57BL/6 (H-2^b) and BALB/c (H-2^d) mice were used as the allogeneic and syngeneic donor mice, respectively. Moreover, female BALB/c mice were used as recipient mice. The recipient mice were conditioned by a single dose (8 Gy) of total body irradiation (TBI). The recipient mice were randomly divided into 7 groups: TBI group, Syn-BMST control group, post-Syn-BMST G-CSF administration (Syn-BMST+G-CSF)group, allo-BMT control group, post-allo-BMT G-CSF administration (allo-BMT+G-CSF)group, allo-BMST control group and post-allo-BMST G-CSF administration (allo-BMST+G-CSF) group. The mice in control groups and G-CSF administration groups were subcutaneous injected with 0.1 mL normal saline (NS) and 0.1 mL NS containing 2 μg G-CSF per day from 1st day, respectively. The effect of G-CSF on aGVHD was evaluated by clinical manifestations and pathological changes, as well as survival time of the mice in different groups. The serum levels of IL-2, IL-4, IFN-γ and TNF-α in allo-BMST and allo-BMST+G-CSF groups were detected by ELISA at 10th day. Flow cytometry was used to analyze the immunophenotypes of splenocytes at 10th day. RESULTS: The mice in TBI group were all died for hematologic failure on 9~15 d after TBI. No effect of G-CSF on the survival of the mice underwent Syn-BMST and transplantation of single allogeneic marrow cells was observed. The mean survival days in allo-BMST group and allo-BMST+G-CSF group were (34.8±4.5) d and (19.8±6.1) d'respectively (P<0.01). Moreover, post-transplant administration of G-CSF increased the spleen total nucleated cells count (SpTNC), NK cells subset, and DC1/DC2 ratio in the spleen with over 99% of donor chimerism rate at 10th day. No difference in the levels of serum IL-2, IL-4, IFN-γ and TNF-α between the 2 group at 10th day was found. CONCLUSION: The administration of G-CSF after allo-BMST significantly aggravates mouse aGVHD. The expansion of NK cells stimulated by G-CSF may be involved in the mechanism of generating alloreactivity against host cells. These results imply there may be potential risk of evoking or aggravating acute GVHD if G-CSF is administered in the early stage of clinical allo-HSCT.     

Experimental study on anti-endotoxin activity of a tetrahydropyrimidine derivative,ZL-5015

AIM: To investigate the protective effect of 1, 3-dicyclopentyl-1, 2, 3, 6-tetrahydropyrimidine-4, 5-dicarboxylic acid diethyl ester (ZL-5015) on lethal endotoxin-challenged mice and to explore the underlying mechanism. METHODS: Mouse model of lethal endotoxin challenge and endotoxemia were established by intraperitoneal administration of lipopolysaccharide (LPS) at a dose of 70 mg/kg to the C57BL/6J mice. Mouse peritoneal macrophages stimulated with LPS (10 mg/L) were used as an in vitro inflammatory model. The levels of interleukin-1β (IL-1β), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was used to evaluate the mRNA expression of the cytokines. RESULTS: Prophylactic treatment of the mice with ZL-5015 (100 and 200 mg/kg, ig) slightly increased the survival rate, extended the survival time, decreased the serum levels of IL-1β and TNF-α, and increased the serum level of IL-10 in the early stage of endotoxemia as compared with model group. The results of in vitro study demonstrated that treatment of the endotoxin-stimulated mouse peritoneal macrophages with ZL-5015 (10, 20 and 40 μmol/L) inhibited the expression of IL-1β and TNF-α at both mRNA and protein levels but promoted the expression of IL-10 at both mRNA and protein levels. CONCLUSION: The tetrahydropyrimidine derivative ZL-5015 shows a moderate anti-endotoxin effect by increasing the survival rate and extending the survival time of the mice challenged by endotoxin, which may result from inhibition of the expression of pro-inflammatory cytokines such as IL-1β and TNF-α, and promotion of the expression of anti-inflammatory cytokine IL-10.     

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     

Effects of Radix Angelicae Sinensis on changes of IL-6, TNF-α and bFGF in renal ischemia/reperfusion injury in rabbits

AIM: To determine the effect of Radix Angelicae Sinensis(RAS) on renal ischemia/reperfusion injury in rabbits and to explore its mechanism. METHODS: Twenty-five rabbits were divided randomly into the sham operated group(Control group), renal ischemia/reperfusion injury group(IR group) and RAS+IR group. At the time point of reperfusion 48 h after renal ischemia 1 h, the renal tissue were observed by electron-microscope and the contents of creatinine(Cr) in serum, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)and basic fibroblast growth factor(bFGF) in the renal tissue were measured. RESULTS: A remarkably degenerative changes in renal tissue were showed under electronmicroscope in IR group, but the changes in RAS+IR group were slight. The contents of Cr, TNF-α and IL-6 in IR group were higher than those in Control group, these parameters in RAS+IR group were lower than those in IR group, the difference between these groups was significant(P< 0.05 or P< 0.01). At the same time, the content of bFGF in IR group was lower than that in Control group(P< 0.01), while the content of bFGF in RAS+IR group was higher than that in IR group(P< 0.01) and Control group(P< 0.05). CONCLUSION: RAS has an effect of alleviating the renal ischemia/reperfusion injury by modulating the production or release of TNF-α, IL-6 and bFGF.     

Effect of IFN-γ inhalation on the cellular immunity of the lungs

AIM:To study the effect of IFN-γ inhalation on the anti-infection ability of the lungs in the immunocompromised host. METHODS:The immunological factors in the immunocompromised rats and the immunocompromised rats administrated IFN-γ via aerosol were investigated after 1, 3, 7 days when they were injected Candida albicans via tracheal. The Canidda albicans count of the left lung was also determined after 7 days when injecting pathogen. RESULTS:The Canidda albicans count of the left lung in IFN-γ group was significantly less than that of control group. The phagocyting and bactericidal percentages, Ia antigen expression percentages, the levels of TNF-α, IL-1β and IL-6 in the culture supernatant of the AM, the activity of IFN-γ and TNF-α in BALF (except the TNF-α on 7 th day) in IFN-γ group were markedly higher than those in control group. The expression of IFN-γ and IL-1β pulmonary tissues in IFN-γ group was higher than that in control group. The expression of TNF-α in IFN-γ group was less than that in control group. The expression of IL-6 was no changes between two groups. The levels of IFN-γ, IL-1β and IL-6 in the blood (except IL-1β on 3 rd day), and the killing ability of the lymphocytes in blood had no difference between two groups. CONCLUSION:Administration of IFN-γ via aerosol obviously enhanced the anti-infection ability of the lungs in the immunocompromised host, but has no influence on the whole body cellular immunity.     

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.     

Effects of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages

AIM: To study the effect of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages. METHODS: Mouse macrophage cell line J774.1 cells were cultured with LPS and liposome-mediated oligodeoxynucleotides, and the levels of TNF-α and IL-6 measured in the different culture supernatant by enzyme linked immunosorbent assay. RNA was extracted from macrophages, and the mRNA expression of TNF-α and IL-6 in macrophages was observed by RT-PCR. RESULTS: NF-κB "decoy" oligodeoxynucleotides decreased the expression of TNF-α and IL-6 in LPS-induced macrophages and inhibited generation of TNF-α and IL-6. The level of TNF-α and IL-6 did not change in control group. CONCLUSIONS: NF-κB "decoy" oligodeoxynucleotides inhibit the expression of TNF-α and IL-6 in LPS-induced macrophages, which is probably due to the specific inhibition of activated NF-κB binding sites .     

Effects of chronic Clonorchis sinensis infestation on inflammatory reaction of macrophages

AIM：To evaluate the immune state in rats with chronic Clonorchis sinesis (Cs) infestation by investigating the effects of Cs on macrophage polarization and inflammatory reactions. METHODS：Sprague-Dawley rats were used in the study. Chronic Cs infestation model was reproduced by intragastric perfusion with Cs eggs. Twenty rats were randomly divided into normal group (n=10) and Cs infestation group (n=10). The serum levels of interleukin (IL-4) and IL-10, tumor necrosis factor α(TNF-α) and interferon γ (IFN-γ) were detected by ELISA. The macrophages were harvested by peritoneal lavage. The differentiation proportion of M1 and M2 macrophages were detected by flow cytometry. The macrophages were divided into control group, normal group and chronic Cs infestation group according to the sources of macrophages. The levels of TNF-α and IL-10 in the culture supernatants were detected by ELISA at 0, 2, 12 and 24 h after lipopolysaccharide (LPS, 10 μg/L) stimulation in vitro. RESULTS：Compared with normal group, chronic Cs infestation increased the serum levels of TNF-α, IFN-γ, IL-4 and IL-10. The differentiation proportion of M1 detected by flow cytometry was 92.1% in normal group and that of M2 macrophages was 93.8% in Cs infestation group. The levels TNF-α and IL-10 in culture supernatants were increased at 2~24 h after LPS stimulation both in normal group and Cs infestation group, but the levels of TNF-α were lower in chronic Cs infestation group than that in normal group at 2 h,12 h and 24 h after LPS stimulation. The level of anti-inflammatory cytokine IL-10 was higher in Cs infestation group than that in normal group at 2 h, 12 h and 24 h after LPS stimulation. CONCLUSION：Chronic Cs infestation increases the serum levels of both pro-inflammatory cytokines and anti-inflammatory cytokines, thus inducing the polarization of M2 macrophages. The macrophages derived from chronic Cs-infected rats produce tolerance in the inflammatory process against LPS in vitro.     

Effects of metformin combined with Ge Xia Zhu Yu decoction on TLR4/NF-κB signaling pathway in rats with polycystic ovary and insulin resis-tance

AIM: To study the effect of metformin (Met) combinated with Ge Xia Zhu Yu decoction on Toll-like receptor-4 (TLR-4)/nuclear factor-κB (NF-κB) signaling pathway in the rats with polycystic ovary syndrome (PCOS) and insulin resistance induced by dehydroepiandrosterone, and to explore the mechanisms. METHODS: PCOS rats (after induced by dehydroepiandrosterone, n=110) were randomly divided into 3 groups:model group (30 rats), Met treatment group (40 rats) and Met combinated with Ge Xia Zhu Yu decoction treatment (combination) group (40 rats). The rats in model group were given the same volume of 0.9% sodium chloride daily by gavage. The rats in Met group were given Met (270 mg·kg^-1·d^-1) by gavage. The rats in combination group were given Met (270 mg·kg^-1·d^-1) and Ge Xia Zhu Yu decoction (34.5 mg·kg^-1·d^-1) by gavage. All rats were continuously intervened for 28 d. After the intervention, blood glucose[fasting plasma glucose (FPG) and 2-hour postprandial blood glucose (2hPBG)] was measured. The mRNA expression levels of follicular epithelial NF-κB, TLR-4 and oxidized low-density lipoprotein (ox-LDL) were detected by RT-PCR. The serum levels of inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were also detected by ELISA. RESULTS: After the intervention, FPG, 2hPBG, and serum levels of IL-6, TNF-α and CRP in Met group and combination group were lower than those in model group (P<0.05), and those in combination group were lower than those in Met group (P<0.05). Meanwhile, the mRNA expression levels of follicular epithelial NF-κB, TLR-4 and ox-LDL in Met group and combination group were lower than those in model group (P<0.05), and those in combination group were lower than those in Met group (P<0.05). CONCLUSION: Treatment with Met combined with Ge Xia Zhu Yu decoction improves insulin resistance in PCOS rats by decreasing the levels of inflammatory factors in serum and epithelial cells of ovary and inhibiting the expression of NF-κB, TLR-4 and ox-LDL in epithelial tissue of ovary.