Research advance on the role of pro-inflammatory cytokines in sepsis / 中华危重病急救医学

Sepsis, a series of pathophysiological abnormalities caused by infection, is also one of the most important factors of death and disability in infected patients all over the world, so it has always been the focus of the medical community. Cytokines are small molecule proteins secreted by cells with biological activity, involved in the immune and inflammatory regulation of sepsis. Many studies using cytokine targeting to treat sepsis have achieved beneficial effects, and the level of cytokines is also believed to be related to the development, severity of sepsis, so they are reliable biomarkers of sepsis. Among them, pro-inflammatory cytokines such as interferon-β (IFN-β) and interleukins (IL-1β, IL-3, IL-6, and IL-7) are the focus of the discussion in this review. IFN-β and IL-1β are double-sided in the treatment of sepsis, namely early low-dose treatment can reduce sepsis by restoring the function of immune cells and play a protective effect, but they are also related to severe inflammatory response of sepsis and can aggravate the mortality of sepsis patients. IL-3 and IL-6 focus more on enhancing inflammatory factors and play a damage role. IL-7 mainly participates in immune regulation, promoting lymphocyte activation and protecting sepsis.

TLR8 in the Trigeminal Ganglion Contributes to the Maintenance of Trigeminal Neuropathic Pain in Mice / 神经科学通报·英文版

Trigeminal neuropathic pain (TNP) is a significant health problem but the involved mechanism has not been completely elucidated. Toll-like receptors (TLRs) have recently been demonstrated to be expressed in the dorsal root ganglion and involved in chronic pain. Here, we show that TLR8 was persistently increased in the trigeminal ganglion (TG) neurons in model of TNP induced by partial infraorbital nerve ligation (pIONL). In addition, deletion or knockdown of Tlr8 in the TG attenuated pIONL-induced mechanical allodynia, reduced the activation of ERK and p38-MAPK, and decreased the expression of pro-inflammatory cytokines in the TG. Furthermore, intra-TG injection of the TLR8 agonist VTX-2337 induced pain hypersensitivity. VTX-2337 also increased the intracellular Ca

The pathogenetic role of macrophage polarization in the necrotizing enterocolitis / 中国小儿急救医学

Objective:To investigate the effect of macrophage polarization in the pathogenesis of necrotizing enterocolitis(NEC).Methods:C57BL/6 mice were chose to construct the NEC model.The preterm pups were randomly assigned into the control group( n=10) and the NEC group( n=19). The pups in the control group were breastfed by mothers while the NEC group were treated with hypoxia, hypothermia, hypertonic feeding and lipopolysaccharide treatment.The intestinal tissues from the lower part of duodenum to the colon were collected after the pups were born after 96 hours.HE staining was used to observe the intestinal histological structure.Intestinal mucosal permeability was detected by the measurement of concentration of FD70 in plasma after gastric gavage.The expression of Pan-keratin of intestinal epithelium was detected by immunofluorescence histochemistry.Enterocyte apoptosis was detected by TUNEL staining.The expression of CD86 and CD206 protein were determined by western blotting and the percentage of M1 and M2 macrophages was calculated by flow cytometry.RT-PCR was used to detect the expression of mRNA levels of granulocyte-macrophage colony stimulating factor, interleukin(IL)-6, IL-10 and IL-12. Results:Compared with the control group, the pups in the NEC group had low survival rate(100.0% vs.36.8%), different level of intestinal injury, incomplete integrity of intestinal epithelium, increased mucosal permeability(1.53±0.80 vs.14.32±1.27, P<0.05)and enterocyte apoptosis(1.9%±1.1% vs.7.6%±2.6%, P<0.05). Western blotting showed that the expression of CD86 protein(1.00±0.01 vs.1.50±0.10, P<0.05) increased while CD206 protein decreased(1.00±0.01 vs.0.60±0.05, P<0.05) in the NEC group.Flow cytometry showed that the percentage of CD68 + CD86 + M1 macrophages increased(1.90%±0.19% vs.10.20%±0.38%, P<0.05) while the CD68 + CD206 + M2 macrophages decreased(5.8%±0.33% vs.3.7%±0.56%, P<0.05) in the NEC group.The expression of the mRNA levels of pro-inflammatory cytokines, granulocyte-macrophage colony stimulating factor(1.00±0.05 vs.1.83±0.17, P<0.05), IL-6 (1.00±0.13 vs.2.00±0.58, P<0.05) and IL-12(1.00±0.05 vs.1.49±0.22, P<0.05) increased and the anti-inflammatory cytokine IL-10(1.00±0.22 vs.0.09±0.01, P<0.05) decreased. Conclusion:Polarization of macrophages towards the pro-inflammatory M1 subtype plays an important role in the pathogenesis of NEC.

Mechanism of effect of IL-17C on survival of kidney graft in mice / 器官移植

Objective To investigate the effect and mechanism of interleukin (IL)-17C in mice undergoing kidney transplantation. Methods The life-supporting kidney transplantation mice models were established using Balb/c (H-2Kd) mice as the donors, IL-17C gene knock out (IL-17CKO) mice (knockout group) and C57BL/6J(H-2Kb) mice (wild group) were chosen as the recipients. The postoperative body mass and survival time of mice were statistically compared between two groups. Pathological examination of the kidney graft was performed by using hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining. The expression levels of granzyme B, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-6 and IL-1β messenger ribonucleic acid (mRNA) in the kidney graft tissue were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). The proportion of inflammatory cell infiltration in the kidney graft tissue was detected by flow cytometry. Results In the knockout group, the survival time of mice after kidney transplantation was significantly shorter than that of the wild mice (P=0.031). The body mass was more evidently decreased in the knockout group with no statistical significance from that in the wild group. Pathological examination demonstrated that the kidney graft injury in the knockout group was significantly worse than that in the wild group. The mRNA expression levels of granzyme B, IFN-γ, TNF-α, IL-6 mRNA in the knockout group were significantly up-regulated compared with those in the wild group (all P < 0.01). The mRNA expression level of IL-1β showed a decreasing trend with no statistical significance (P=0.16). Flow cytometry analysis revealed that the infiltration of CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6Chi monocyte in the kidney graft of knockout mice was significantly higher compared with that of the wild mice (P < 0.05, P < 0.01), whereas the infiltration of CD45+Ly6ChiF4/80+ macrophage did not significantly differ between two groups (P > 0.05). Conclusions IL-17C participates in the regulation of inflammatory response after kidney transplantation. It can alleviate acute rejection and improve the survival of kidney graft by down-regulating the expression of pro-inflammatory cytokines and infiltration of inflammatory cells.

Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of IL-1α, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of IL-1α, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

Correlation between clinical findings, mast cell count and interleukin 31 immunostaining in the skin of dogs with atopic dermatitis / Correlação entre achados clínicos, contagem de mastócitos e imunomarcação de interleucina 31 na pele de cães com dermatite atópica

ABSTRACT: In this study the correlation between the clinical score, mast cell count and interleukin 31 (IL-31) immunostaining in the skin of dogs with atopic dermatitis was determined. A total of 31 dogs of different breeds, from one to eight years of age, were chosen for the study. The 20 females and 11 males were categorized based on the CADESI-4 system, as having discrete, moderate or marked atopic dermatitis. Skin samples were collected from the axillary and interdigital regions and stained with hematoxylin and eosin for cytohistomorphological analyses and toluidine blue to evaluate the mast cell counts, and immunohistochemistry for the IL-31 immunostaining. Animals revealing higher atopic dermatitis scores had greater numbers of mast cells and IL-31 immunolabeled cells. More numbers of cells immunolabeled for IL-31 were evident in the axillary skin compared with the interdigital skin in dogs having this condition. A correlation was identified between the clinical scores and mast cell numbers in the interdigital region, as well as between the clinical scores and number of cells immunolabeled for IL-31 in the axillary area. A correlation was also reported between the mast cell numbers and IL-31 immunolabeled cells only in the axillary skin, and none in the interdigital regions. It was thus concluded that the mast cells and IL-31 are involved in the pathogenesis of the canine atopic dermatitis (CAD), as well as lymphocytes and plasma cells. It was also observed that the higher the degree of clinical severity of the disease, the more the numbers of mast cells and IL-31 in the skin of those animals suffering from CAD, which implies the influence of these immunological constituents on the genesis of pruritus and disease progression.

RESUMO: Este estudo avaliou a correlação entre o escore clínico, a contagem de mastócitos e a imunomarcação de interleucina 31 (IL-31) na pele de cães com dermatite atópica. Foram selecionados 31 cães de diferentes raças, com idade entre um e oito anos, sendo 20 fêmeas e 11 machos, divididos em discretamente, moderadamente e acentuadamente acometidos por dermatite atópica segundo o sistema CADESI-4. Amostras da pele das regiões axilar e interdigital foram colhidas e submetidas às colorações de hematoxilina e eosina para a avaliação cito-histomorfológica e azul de toluidina para a contagem de mastócitos, bem como a técnica de imunoistoquímica para a imunomarcação de IL-31. Os animais com maior escore de dermatite atópica apresentaram maior número de mastócitos e de células imunomarcadas para IL-31. Houve maior número de células imunomarcadas para IL-31 na pele da axila em relação à interdigital nos cães com a doença. Foi constatada correlação entre o escore clínico e a quantidade de mastócitos no interdígito, bem como entre o escore clínico e a quantidade de células imunomarcadas para IL-31 na axila. Também foi verificada correlação entre a quantidade de mastócitos e células imunomarcadas para IL-31 na pele da região axilar, mas não da interdigital. Conclui-se que mastócitos e a IL-31 estão envolvidos na patogenia da DAC, assim como linfócitos e plasmócitos. Também, quanto maior o grau de severidade clínica da doença, maior a quantidade de mastócitos e IL-31 na pele dos animais com DAC, o que remete à influência desses componentes imunológicos na gênese do prurido e progressão da doença.

IL-17A-Producing Foxp3⁺ Regulatory T Cells and Human Diseases

CD4⁺Foxp3⁺ regulatory T (Treg) cells play major roles in immune homeostasis. While CD4⁺Foxp3⁺ Treg cells act to suppress other immune effector cells, there is growing evidence that they also produce pro-inflammatory cytokines, such as IL-17A, in inflammatory conditions. The pro-inflammatory cytokine milieu, toll-like receptor (TLR) signaling, and specific transcription factors are important for the production of IL-17A by CD4⁺Foxp3⁺ Treg cells. In particular, IL-17A-producing CD4⁺Foxp3⁺ Treg cells express RORγt, the T helper (Th) 17-specific transcription factor, in addition to Foxp3. IL-17A-producing CD4⁺Foxp3⁺ Treg cells are also involved in the pathogenesis of various diseases. Here we review the mechanisms underlying the induction of IL-17A-producing CD4⁺Foxp3⁺ Treg cells and the roles of these cells in human disease.

Combined approaches using adverse outcome pathways and big data to find potential diseases associated with humidifier disinfectant

According to previous survey, about two million of people were expected to suffer from toxic effects due to humidifier disinfectant (HD), regardless of healing or not. Extremely small group are recognized as HDs’ victims. Up to now, previous research tried to focus on interstitial fibrosis on terminal bronchiole because it is specific finding, compared with other diseases. To figure out overall effects from HDs, we recommend adverse outcome pathways (AOPs) as new approach. Reactive oxygen species (ROS) generation, decreased T-cell and pro-inflammatory cytokine release from macrophage could be key events between the exposure to HDs and diseases. ROS generation, decreased cell and pro-inflammatory cytokine release from macrophage could be cause of interstitial fibrosis, pneumonia and many other diseases such as asthma, allergic rhinitis, allergic dermatitis, fetal death, premature baby, autoimmune disease, hepatic toxicity, renal toxicity, cancer, and so on. We predict potential disease candidate by AOPs. We can validate the real risk of the adverse outcome by epidemiologic and toxicologic study using big data such as National Health Insurance data and AOPs knowledge base. Application of these kinds of new methods can find the potential disease list from the exposure to HD.

Combined approaches using adverse outcome pathways and big data to find potential diseasesassociated with humidifier disinfectant

According to previous survey, about two million of people were expected to suffer from toxic effects due to humidifier disinfectant (HD), regardless of healing or not. Extremely small group are recognized as HDs’ victims. Up to now, previous research tried to focus on interstitial fibrosis on terminal bronchiole because it is specific finding, compared with other diseases. To figure out overall effects from HDs, we recommend adverse outcome pathways (AOPs) as new approach. Reactive oxygen species (ROS) generation, decreased T-cell and pro-inflammatory cytokine release from macrophage could be key events between the exposure to HDs and diseases. ROS generation, decreased cell and pro-inflammatory cytokine release from macrophage could be cause of interstitial fibrosis, pneumonia and many other diseases such as asthma, allergic rhinitis, allergic dermatitis, fetal death, premature baby, autoimmune disease, hepatic toxicity, renal toxicity, cancer, and so on. We predict potential disease candidate by AOPs. We can validate the real risk of the adverse outcome by epidemiologic and toxicologic study using big data such as National Health Insurance data and AOPs knowledge base. Application of these kinds of new methods can find the potential disease list from the exposure to HD.

Effect of excretory/secretory protein of Trichinella spiralis adult worm on CLP-induced sepsis in mice / 中国血吸虫病防治杂志

Objective To observe the effect of excretory/secretory products from Trichinella spiralis adult worms(AES)on cecal ligation and puncture(CLP)?induced sepsis in mice. Methods Forty?eight BALB/c mice were randomly divided into 3 groups:a sham operation group(PBS+sham group,Group A),a CLP?induced sepsis group(PBS+CLP group,Group B)and an AES treatment group(AES+ CLP group,Group C). The mice of each group were intraperitoneally injected with 25 μg of AES or PBS only as a control in a total volume of 200μl. Eight mice from each group were selected randomly for survival analy?sis of 96 hours. The other 8 mice in each group were observed for pathological changes in the lung,liver and kidney tissues by HE staining 12 h after CLP,and then determined for the detection of cytokines including TNF?α,IL?1β,IL?6,IL?10 and TGF? βin the sera by ELISA. Results The difference among the survival rates of mice in the 3 groups was statistically significant (χ2=21.16,P<0.05). Compared to Group A(100%),the survival rate of mice in Group B(0)decreased significantly(P<0.05),and also the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group B increased signifi?cantly after CLP. Compared with the mice in group B,the survival rate of those in Group C(70%)increased significantly(P<0.05),and the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group C decreased significantly after the treatment with AES. The differences among the levels of pro?inflammatory cytokines TNF?α(F=27.11,P<0.05),IL?1β(F=18.75,P<0.05)and IL?6(F=100.93,P<0.05)in the sera of the mice in the three groups were statistically signifi?cant. Compared with the mice in Group A,the levels of the 3 cytokines of those in Group B increased significantly(all P <0.05). However,after the treatment with AES,the levels of the pro?inflammatory cytokines of those in Group C decreased signifi?cantly(all P<0.05). The differences among the levels of immunoregulatory cytokines IL?10(F=10.88,P<0.05)and TGF?β(F=11.37,P<0.05)in the sera of the mice in the three groups were also statistically significant. Compared with the mice in Group B,the levels of IL?10 and TGF?β of those in Group C were higher after treatment with AES(both P<0.05). Conclu?sion T. spiralis AES has a therapeutic potential for alleviating sepsis induced by CLP in mice.

Serum-and glucocorticoid-regulated kinase 1 suppresses inflammatory responses mediated by toll like receptor 4 via nuclear factor-κB / 中华传染病杂志

Objective To investigate the role of serum and glucocorticoid regulated protein kinase (SGK) 1 in the inflammatory responses mediated by toll like receptors.Methods Mice were injected with lipopolysaccharide (LPS,1 mg/kg) 2 h after the pretreatment of EMD638683 (10 mg/kg) or phosphate buffered saline (PBS) as control.At the time points of 3 and 24 h,pro-inflammatory cytokines (interleukin [IL]-6,IL-12 and tumor necrosis factor [TNF]-α) in serum were measured using enzymelinked immunosorbent assay (ELISA).Livers and lung were harvested at 6 h and 24 h after the injection of LPS,embedded by optimum cutting temperature (OCT) and then stained with hematoxylin and eosin (HE).Peripheral blood mononuelear cell (PBMC) were isolated and stimulated by LPS with or without the pretreatment of EMD or LY294002.Cytokines (IL-6,IL-12 and TNF-α) were measured using ELISA.IKKα/β,IKBα and nuclear factor (NF)-κB p65 were detected by Western bolt.Data were analyzed by one way analysis of variance.Results In the model of LPS-induced endotoxin sepsis,inhibition of SGK1 induced secretion of pro-inflammatory cytokine (IL-6 [t=3.007,P＜0.05],IL-12[t=4.413,P＜0.05] and TNF-α[t=5.403,P＜0.05]),increased inflammatory cells infikration into the liver and lung within 6 h,and induced serious multiple organ damage with collapse of alveoli and fatty degeneration of liver.After 24 h,pharmacological inhibition of SGK1 with EMD638683 increased proinflammatory cytokine (IL-6 [t=18.540,P＜0.01],IL-12[t=16.520,P＜0.01] and TNF-α[t=34.880,P＜0.01]) production in human PBMC upon LPS stimulation and inhibited the phosphorylation of IKKα/ β/IKBα and nuclear factor (NF)-κB p65.Conclusions SGK1 suppresses the toll like receptor 4 mediated inflammatory responses via NF-κB.

Acrosorium polyneurum Extract Inhibits the LPS-Induced Inflammatory Response by Impairing the MAPK and NF-κB Pathways

Marine algae exhibit broad spectrum anti-bacterial and anti-inflammatory activities. Acrosorium polyneurum (A. polyneurum) is a marine red alga and belongs to the family Delesseriaceae. The present research evaluates the antiinflammatory effects of A. polyneurum extract (APE) on pro-inflammatory cytokine production. APE demonstrated substantial inhibitory effects on production of pro-inflammatory cytokine in bone marrow-derived macrophages (BMDMs). APE pre-treatment in the lipopolysaccharide (LPS)-stimulated BMDMs exhibited a robust inhibitory effect on production of interleukin (IL)-12, IL-6 and tumor necrosis factor (TNF)-α. It revealed a robust inhibitory effect on phosphorylation of ERK1/2, JNK1/2 and p38. APE also showed remarkable inhibitory effect on phosphorylation and degradation of IκBα. Furthermore, APE pre-treatment demonstrated substantial inhibition of LPS-induced production of nitric oxide and inducible nitric oxide synthase. Collectively, these data suggest that APE has a noteworthy anti-inflammatory property and deserve further studies concerning its potential use as a medicinal agent for inflammation-related disorders.

The Changes of Pro- and Anti-inflammatory Cytokines in Both Serum and Gastrocnemius Muscle of Mice after 2-hour Postischemic Reperfusion Injury / 체질인류학회지

This study aimed to investigate the inflammatory changes and their main indicators according to the time-period of postischemic reperfusion injury confirmed by analyzing changes of both pro-inflammatory and anti-inflammatory cytokines in the skeletal muscle and serum. By using 12-week-old male ICR strain mice were grouped into sham control and 8 different time-periods of reperfusion groups (0, 0.5, 1, 2, 4, 8, 16, 24 hours). Left common iliac artery of each mice in the reperfusion group was devascularized by a vascular clamp for 2 hours. Once anesthesia was applied to the experimental animals, blood serum was obtained from right heart atrium on the difference time-period of reperfusion (0-, 0.5-, 1-, 2-, 4-, 8-hour, respectively). Then, tissue fluid was collected in calf muscles (gastrocnemius muscle) after the mice were sacrificed by cervical dislocation. By using these serum and tissue fluids, enzyme-linked immunosorbent assay (ELISA) was used to analyze both pro-inflammatory cytokines (Eotaxin, IFNgamma, IL-1alpha, IL-1beta, IL-2, IL-3, IL-5, IL-6, MCP-1, MDC, MIP-1alpha, RANTES, TARC, TCA-3) and anti-inflammatory cytokines (IL-4, IL-10). Consequently, there were significant differences of pro-inflammatory cytokines levels in the skeletal muscle of 0-hour reperfusion group (p<.05) and those in the serum of 0-, 1-, 2-, 4-, 8-, 16-hour reperfusion groups (p<.05). In the serum of 4-hour reperfusion group, the presence of anti-iflammatory cytokines was significant from other groups (p<.05). By the comparison with the control group, furthermore, pro-inflammatory cytokines in the serum of 2-, 4-, 16-hour reperfusion group and anti-inflammatory cytokines in the serum of 4-hour reperfusion group were considerably different (p<.05). To sum up, changes of cytokine levels according to the time-period of reperfusion were considerably different in the serum rather than the tissue fluids from the skeletal muscle. In particular, IL-6 and MCP-1 in the serum showed higher density in 4- and 16-hour reperfusion groups so that they could be considered as the main indicator of pro-inflammatory cytokines.

Emodin Isolated from Polygoni cuspidati Radix Inhibits TNF-alpha and IL-6 Release by Blockading NF-kappaB and MAP Kinase Pathways in Mast Cells Stimulated with PMA Plus A23187

Emodin, a naturally occurring anthraquinone derivative isolated from Polygoni cuspidati radix, has several beneficial pharmacologic effects, which include anti-cancer, anti-diabetic, and anti-inflammatory activities. In this study, the authors examined the effect of emodin on the production of proinflammatory cytokines, such as, tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, in mouse bone marrow-derived mast cells (BMMCs) stimulated with phorbol 12-myristate 13-acetate (PMA) plus the calcium ionophore A23187. To investigate the mechanism responsible for the regulation of pro-inflammatory cytokine production by emodin, the authors assessed its effects on the activations of transcriptional factor nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinases (MAPKs). Emodin attenuated the nuclear translocation of (NF)-kappaB p65 and its DNA-binding activity by reducing the phosphorylation and degradation of IkappaBalpha and the phosphorylation of IkappaB kinase B (IKK). Furthermore, emodin dose-dependently attenuated the phosphorylations of MAPKs, such as, extracellular signal-regulated kinase 1/2 (ERK1/2), p38 MAP kinase, and the stress-activated protein kinases (SAPK)/c-Jun-N-terminal kinase (JNK). Taken together, the findings of this study suggest that the anti-inflammatory effects of emodin on PMA plus A23187-stimulated BMMCs are mediated via the inhibition of NF-kappaB activation and of the MAPK pathway.

The Soluble Form of the Cellular Prion Protein Enhances Phagocytic Activity and Cytokine Production by Human Monocytes Via Activation of ERK and NF-kappaB

The PrP(C) is expressed in many types of immune cells including monocytes and macrophages, however, its function in immune regulation remains to be elucidated. In the present study, we examined a role for PrP(C) in regulation of monocyte function. Specifically, the effect of a soluble form of PrP(C) was studied in human monocytes. A recombinant fusion protein of soluble human PrP(C) fused with the Fc portion of human IgG1 (designated as soluble PrP(C)-Fc) bound to the cell surface of monocytes, induced differentiation to macrophage-like cells, and enhanced adherence and phagocytic activity. In addition, soluble PrP(C)-Fc stimulated monocytes to produce pro-inflammatory cytokines such as TNF-alpha, IL-1beta, and IL-6. Both ERK and NF-kappaB signaling pathways were activated in soluble PrP(C)-treated monocytes, and inhibitors of either pathway abrogated monocyte adherence and cytokine production. Taken together, we conclude that soluble PrP(C)-Fc enhanced adherence, phagocytosis, and cytokine production of monocytes via activation of the ERK and NF-kappaB signaling pathways.

Anti-inflammatory Activity of Carpinus tschonoskii Leaves Extract in R848-stimulated Bone Marrow-derived Macrophages and Dendritic Cells

The present study aims to evaluate the anti-inflammatory effect of methanol extract from leaves of Carpinus tschonoskii (CE) on R848-stimulated primary bone marrow-derived macrophages (BMDMs) and dendritic cells (BMDCs). Primary BMDMs and BMDCs were used for pro-inflammatory cytokine production. Human embryonic kidney cell line 293T (HEK293T) was used to access NF-kappaB activity. In all cases, R848 was used to stimulate the cells. The CE (0~150 microg/ml) was treated to BMDMs, BMDCs, and HEK293T cells. CE pre-treatment in R848-stimulated BMDMs and BMDCs showed a dose-dependent inhibitory effect on pro-inflammatory cytokine (e.g., IL-12 p40, IL-6, and TNF-alpha) production as compared to non-treated controls. In NF-kappaB reporter gene assay, the CE pre-treatment inhibited NF-kappaB-dependent luciferase activity in a dose-dependent manner. Overall, our findings suggest that CE has significant inhibitory effect on pro-inflammatory cytokine production and deserve further studies concerning potentials of CE for medicinal uses.

Effects of intrathecally coadministered dexamethasone and spironolactone on pain behaviors in a rat model of radicular pain / 中华行为医学与脑科学杂志

Objective To investigate the effects of intrathecally coadministered dexamethasone and spironolactone in trathecally on radicular pain behaviors.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion (CCD) ,48 male SD rats successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into Sham-operation group (Sham group, n= 12),Control group ( C group, n = 12 ), Dexamethasone group ( D group, n = 8 ), Spironolactone group ( S group, n = 8 )and Dexamethasone plus spironolactone group (DS group, n=8).Rats in D group,S group or DS group were intrathecally treated with dexamethasone 4 μg, spironolactone 3 μg or dexamethasone 4 μg plus spironolactone 3 μg twice daily on day 2 ～4 after CCD respectively,while rats in C and Sham group received 10μl 10% alcohol.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before CCD and day 1,4,7,10,14,17 and 21 after CCD.Results Compared with Sham group, both PWMT and PWTL were significantly decreased after CCD surgery on the ipsilateral side(P＜0.01 ＝.Intrathecally administrated with dexamethasone significantly improved pain behaviors (P＜0.01 ＝ and these therapeutic effects lasted up to 10 days after CCD surgery.As with dexamethasone,intrathecal spironolactone also significantly attenuated PWMT (P＜0.01 ＝ and PWTL (P＜0.01 ＝ and the change lasted up to 7 days after CCD surgery.Coadministration spironolactone and dexamethasone exhibited significant synergies( PWMT: ( 13.52 ± 0.72) g vs ( 11.58 ± 1.38 ) g, P ＜0.01; PWTL: ( 19.63 ± 1.68) s vs ( 14.14 ± 1.52) s, P ＜ 0.01 ＝.These effects lasted up to at least 10 days.Conclusion Both dexamethasone and spironolactone intrathecally have therapeutic effects on radicular pain behaviors, combination injection of these two drugs could generate significant synergies.

Naegleria fowleri Lysate Induces Strong Cytopathic Effects and Pro-inflammatory Cytokine Release in Rat Microglial Cells

Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A 51Cr release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-alpha, IL-6, and IL-1beta, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response.

Expression of Pro-inflammatory Cytokines during Distraction Osteogenesis of the Rat Tibia / 대한정형외과학회잡지

PURPOSE: The purposes of this study were to investigate the expression pattern of pro-inflammatory cytokines during distraction osteogenesis and to compare these with expression during simple fracture healing. MATERIALS AND METHODS: Regenerated bones from the rat tibia subjected distraction osteogenesis and simple fracture healing models were harvested over three-week periods. Temporal expressions of mRNA of pro-inflammatory cytokines were investigated by RNase protection assay. Immunohistochemical studies for IL-6 were performed in postoperative day 7 and 9 tissue section specimens. RESULTS: IL-1beta and IL-6 produced detectable signals, while IL-1alpha, TNFalpha and TNFbeta did not. The mRNA expressions of IL-1beta and IL-6 were markedly upregulated on postoperative day 1 and then subsided to the preoperative level. IL-1beta mRNA expression remained the same even when distraction began. However, IL-6 mRNA expression was reactivated during the distraction phase. Immunohistochemical study revealed the expressions of IL-6 not only at the transitional zone of the transchondroid ossification, in young osteoblasts lining newly formed trabeculae and in hematopoietic cells in the marrow but also in primitive mesenchymal cells at the distraction gap. CONCLUSIONS: Distraction strain re-induced IL-6 expression during distraction osteogenesis, which suggests that well-controlled inflammatory reaction might contribute to active new bone formation in distraction osteogenesis.

Pro-inflammatory cytokine expression through NF-kappaB/IkappaB pathway in lung epithelial cells / 결핵

BACKGROUND: The importance of pro-inflammatory cytokines, especially tumor necrosis factor α(TNF-α) and interleukin-1β(IL-1β), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the actiation of NF-κB. However, the relationship between pro-inflammatory cytokine expression and NF-κB/IκB pathway in lung epithelial cells is not clear. METHODS: BEAS-2B, A549, NCI-H719 cells were stimulated with IL-1β or TNF-α at various times, and then IL-8 and TNF-αmRNA expressions were assayed by Northern blot analysis. IL-1β or TNF-α-induced NF-κB activation was assessed by the nuclear translocation of p65 NF-κB subunit. The degradation of IκBα and IκBβ by IL-1βor TNF-α stimulation was assayed by Western blot analysis. The phosphorylation of IκBαwas evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-1β or TNF-α stimulation. The basal level of IKKα expression was evaluated by Western blot analysis. RESULTS: IκBαand IκBβ was repidly degraded after 5 minutes of incubation with IL-1β or TNF-α in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-κB and the induction of IL-8 and TNF-α mRNA expressions were observed by IL-1β or TNF-α stimulation in these cells. In contrast, neither the changes in NF-κB/IκB pathway nor IL-8 and TNF-α mRNA expression was induces by IL-1β or TNF-α stimulation in NCI-H719 cell. IL-1β and TNF-α-induced IκB phoshorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKKα expression was not different between cells. CONCLUSION: NF-κB/IκB pathway plays an important role in the ixpression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-κB/IκB pathway in NCI-H719 cells seems to be due to the defect in the intracellular signal transduction pathway upstream to IKK.