Effects of carbamazepine and metabolites on IL-2, IL-5, IL-6, IL-10 and IFN-γ secretion in epileptic patients: the influence of co-medication

Carbamazepine is a widely used anticonvulsive agent. Its metabolic pathway leads not only to the major active metabolite, carbamazepine-10,11-epoxide, but also to minor terminal metabolites such as iminostilbene and acridine. Carbamazepine is usually well-tolerated, but it may lead to rare, but serious, hypersensitive reactions associated with hypereosinophilia. The mechanisms of hypersensitivity reactions to carbamazepine are still largely unknown, and the implications of the cell-mediated immune response (Th1 pathway) or the humoral immune response (Th2 pathway) are still not understood in these reactions. It is also unclear whether the parent drug or its subsequent metabolites are the primary trigger agent. In our study, we performed ex vivo experiments to evaluate the stimulation of cytokine secretion by carbamazepine, carbamazepine-10,11-epoxide, iminostilbene and acridine. IL-5, IL-6 and IL-10 were quantified as markers of the Th2 pathway, and IL-2 and IFN-γ were used as markers of the Th1 pathway. Blood samples (n=24) were obtained from epileptic patients routinely treated with carbamazepine alone or co-treated with lamotrigine or valproate. The concentrations of cytokines in the plasma were determined before and after 3 h stimulation with drugs. We found a significantly positive effect of co-treatment with valproate on the basal level of IL-5 (p<0.01) and IL-10 (p<0.05). IL-5 production increased only in blood stimulated with a high level of acridine (33 μM), whereas IL-6 production was less specifically stimulated (p<0.05). Because IL-5 is the most potent stimulating factor of the eosinophils, we suggest that the potential helper effect of valproate and acridine can lead to hypersensitive reactions to carbamazepine in the context of the humoral immune response.     

Generation of TNFα, IFNγ, IL-4, IL-6 and IL-10 in Trichinella spiralis infected mice: effect of the anti-inflammatory compound mimosine

The plant amino acid mimosine has been demonstrated to arrest cell cycle progression in the late G1-phase, and inhibits [3H] thymidine incorporation in cultured fibroblasts. In this study, 10 mice were infected with Trichinella spiralis, a nematode parasite, and treated with the antiinflammatory compound L-mimosine to determine if any alteration in the chronic inflammatory state occurred by investigating the host's immunological response. Mimosine was used at 250 g/bolus for 25 days starting five days before the infection and continuing daily for 35 days then TNF, IFN-, IL-4, IL-6, and IL-10 were determined by ELISA method, after 0, 1, 7, 14, 21, 28, 35 days post-infection, in the serum of treated or untreated animals. When animals with T. spiralis were treated with L-mimosine, inhibition of TNF was observed within 21 days post-infection, compared with the controls (untreated mice). IFN was inhibited only up to the 21^st day, while IL-6 was inhibited up to the 7^th day post-infection and the inhibition of IL-4 was seen mainly at 21^st and 35^th day p.i. Mimosine-treated mice did not statistically affect the secretion of IL-10 (p > 0.05). In healthy animals, the production of cytokines were within the same limits compared with those of non-infected animals treated with L-mimosine. Our studies suggest that mimosine proved to be more effective in inhibiting TNF and IL-6, which are mainly produced by macrophages and less effective in inhibiting IL-4, which is produced by T-cells.     

IL-17 ,IL-6 ,IL-1β与儿童原发性肾小球疾病的相关性研究

目的探讨T h17细胞相关细胞因子IL-17、IL-6 ,IL-1β等在原发性肾小球疾病的表达水平及其意义 ,以期为诊治原发性肾小球疾病找到新的治疗靶点提供实验室依据.方法将研究对象分为实验组和对照组 ,实验组为原发性肾小球疾病患儿30例 ,取其外周血 ;并收集10例健康体检儿童外周血作为正常对照组.通过荧光定量PCR检测对IL-17 mRNA在肾小球疾病患儿及健康儿童外周血中的表达进行分析 ;酶联免疫吸附试验(ELISA )分别检测血浆中IL-17、IL-1β以及IL-6的表达.结果 (1)原发性肾小球疾病儿童IL-17mRNA表达水平显著高于正常对照组(P<0.05 ).(2)肾组织内 ,实验组患儿血浆中IL-17、IL-1β以及IL-6的表达明显高于正常对照组(P<0.05).结论Th17细胞相关炎性因子在原发性肾小球疾病的发病机制中起着重要作用 ,特异性针对Th17细胞相关炎性因子的治疗可能是治疗原发性肾小球疾病的新靶点.     

Role of TRPM2 ion channel,an oxidative stress sensor,in hyperglycemiainduced pro-inflammaotry IL-1β in human monocytes

OBJECTIVE To investigate the role of transient receptor potential melastatin 2(TRPM2),a calcium-permeable non-selective cation channel which acts as an oxidative stress sensor,in mediating the production of pro-inflammatory IL-1βin high glucose condition.METHODS Human pro-monocytic leukemia cell U937 was purchased from ATCC and cultured in RPMI 1640(Life Technologies).Prior to high glucose(HG)stimulation,U937 cells were cultured in medium with glucose 5.5mmol·L-1 for 48 h.The cells were then incubated in high glucose concentration(30mmol·L-1)or mannitol(30mmol·L-1)for 48 h.The protein expression of TRPM2 and the production of human IL-1β were evaluated by ELISA.TRPM2 inhibitors(DPQ and AMP)and TRPM2 siRNAs were employed to further investigate the role of TRPM2 in HG-induced IL-1β production.RESULTS The TRPM2 protein expression was significantly up-regulated by 2-folds in U937 cells after the treatment of high glucose(30mmol·L-1 for 48h)(P<0.01).The production of IL-1β in U937 was also significantly increased by HG treatment and was time-and dose-dependent(10,20 or 30mmol·L-1 glucose for 24,48 or 72h)(P<0.01).The HG-induced IL-1β production in U937 could be abolished by using TRPM2 inhibitors DPQ(100μmol·L-1 for 45min)and AMP(100μmol·L-1 for 45 min)as well as by the transfection of TRPM2siRNAs(60nmol·L-1).CONCLUSION High glucose condition(such as in diabetes)might mediate pro-inflammatory environments via the modulation of TRPM2 channels on immune cells.     

Plasma IL-2, NK, IFN-γ, and C3 in male workers exposed to traffic pollutants

The aim of the study is to evaluate if the occupational exposure to urban stressors could cause alterations in interleukin-2 (IL-2), NK, interferon-γ (IFN-γ) and C3 plasma levels in male traffic police officers compared to controls. After excluding the principal confounding factors, 108 traffic police officers were matched with 108 controls by age, working life, habitual consumption of alcohol and spirits. IL-2 mean levels were significantly higher in traffic police officers compared to controls (p=0.04). The distribution of IL-2 values in traffic police officers and in controls was significant (p=0.01). The distribution of NK value percentage in traffic police officers and in controls was significant (p=0.000). IFN-γ and C3 mean levels were not significant in traffic police officers compared to controls. Our results suggest that the occupational chronic exposure to low doses of urban stressors could affect NK and IL-2 plasma concentrations in traffic police officers of male sex.     

The influence of mianserin on TNF-α, IL-6 and IL-10 serum levels in rats under chronic mild stress

BackgroundAntidepressants are known to affect the immunological system through mechanisms which are not completely understood. The aim of the present study was to evaluate the effect of the atypical antidepressant mianserin on the levels of tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the blood of rats in an experimental model of depression.MethodsMale Wistar rats were subjected to chronic mild stress (CMS) according to Willner's method for 6 weeks. Following the development of anhedonia, the stressed and control rats (non-stressed animals) were treated with mianserin (10 mg/kg ip, twice daily) for three weeks. On the last day of the experiment, a lipopolysaccharide (LPS, 100 μg/kg ip) was injected to mianserin- or vehicle-treated rats. TNFα, IL-6 and IL-10 levels in the blood of the rats were assayed using ELISA methods.ResultsThe results indicated a significantly increased TNFα level in stressed animals when compared with the non-stressed (control) group. The levels of IL-6 and IL-10 were also elevated, especially after LPS administration. Treatment with mianserin resulted in a significant lowering of TNFα and IL-6 levels both in LPS-treated and LPS-untreated animals. There was also a decrease in IL-10 concentration in LPS-treated stressed animals.ConclusionsThe results confirm an increase in proinflammatory cytokines in the blood of rats with experimentally induced depression and show the protective role of the activity of mianserin on the cytokine levels, expressed in a lowering of TNFα and IL-6 levels in stressed animals, and of IL-10 levels after LPS administration.     

Ursodeoxycholic acid inhibits pro-inflammatory repertoires,IL-1β and nitric oxide in rat microglia

Ursodeoxycholic acid (UDCA) is a non-toxic, hydrophilic bile acid in widespread clinical use mainly for acute and chronic liver disease. Recently, treatment with UDCA in hepatic graft-versus-host disease has been given in immunosuppressive therapy for improvement of the biochemical markers of cholestasis. Moreover, it has been reported that UDCA possesses immunomodulatory effects by the suppression of cytokine production. In the present study, we hypothesized that UDCA may inhibit the production of the pro-inflammatory cytokine, IL-1beta, and nitric oxide (NO) in microglia. In the study, we found that 100 microg/mL UDCA effectively inhibited these two pro-inflammatory factors at 24 h and 48 h, compared to the Abeta42-pretreated groups. These results were compared with the LPS+UDCA group to confirm the UDCA effect. As microglia can be activated by several stimulants, such as Abeta42, in Alzheimers brain and can release those inflammatory factors, the ability to inhibit or at least decrease the production of IL-1beta and NO in Alzheimers disease (AD) is essential. Using RT-PCR, ELISA and the Griess Reagent System, we therefore found that UDCA in Abeta42 pre-treated cultures played a significant role in suppressing the expression or the production of IL-1beta and NO. Similarly, lipopolysaccharide (LPS) did not activate microglia in the presence of UDCA. Moreover, we found that UDCA exhibits a prolonged effect on microglial cells (up to 48 h), which suggests that UDCA may play an important role in chronic cell damage due to this long effect. These results further imply that UDCA could be an important cue in suppressing the microglial activation stimulated by massive Aa peptides in the AD progressing brain.     

脑出血大鼠血清IL-17,IL-23的表达及意义

目的:以脑出血大鼠为研究对象,探讨血清IL-17,IL-23的表达及意义.方法:将90只大鼠随机分成3组:对照组(A组),假手术组(B组),出血组(C组),每组30只.分别设6h,24h,3d,7d,14d五个时间点.用酶联免疫吸附法(ELISA)检测大鼠血清中白细胞介素17(IL-17)、白细胞介素23(IL-23)的表达.结果:对照组与假手术组IL-17,IL-23的表达在各时间点均无统计学差异(P＞0.05).出血组与对照组、假手术组相比、同一时间点IL-17,IL-23的表达均具有统计学差异(P＜0.05).出血组内各时间点IL-17,IL-23的表达相比,均具有统计学差异(P＜0.05).IL-17,IL-23的表达在脑出血后7d高于出血其他时间点,具有统计学差异(P＜0.05).结论:脑出血大鼠发病6h后,外周血中IL-17,IL-23的表达增加;IL-17,IL-23的表达在脑出血后7d高于6h,24h,3d,14d.IL-17,IL-23参与脑出血后的炎症反应.     

Brain IL-1β was involved in reserpine-induced behavioral depression in rats

AIM: To investigate the mechanism of brain interleukin-1β(IL-1β) in reserpine-induced behavioral depression in rats. METHODS: Porsult swim test was used in the measurement of depressive behavior and ELISA was used in measurement of brain IL-1β. RESULTS: Intraperitoneal injection of reserpine (0, 4, 6, and 8 mg/kg, ip) increased floating time in the Porsult swim test in a dose-and time-dependent manner in rats. Intracerebroventricular injection(icv) of IL-1β receptor antagonist (IL-1ra, 6 mg/kg) blocked the increment of floating time in Porsult swim test at 48 and 72 h after reserpine injection, but not at 1 and 24 h after injection. Brain IL-Iβ increased after reserpine treatment in posterior cortex, hippocampus, and hypothalamus. The increase of IL-1β concentration starts at 24 hours after injection of reserpine and reached the peak at 48 h. CONCLUSION: Reserpine induced behavioral depression partially via brain interleukin-1β generation.     

流行性出血热患者血清IL-12,IL-6,TN F浓度检测分析及临床意义

目的观察轻型和重型流行性出血热患者血清中,IL-12,IL-6,TNF的改变及临床意义。方法收集流行性出血热轻型和重型病人四期血液,用酶联免疫方法(ELISA)等方法检测炎症因子,并与正常人进行比较。结果发热期:轻型和重型与对照组比较,IL-12均明显增高(P均<0.05);重型与对照组比较,TNF明显增高(P<0.05)。少尿期:轻型和重型与对照组比较,IL-12明显增高(P均<0.05);轻型和重型与对照组比较,TNF明显增高(P均<0.05)。多尿期:轻型和重型与对照组比较,IL-12明显增高(P<0.01);轻型与对照组比较,IL-6明显增高(P<0.01)。恢复期:轻型和重型与对照组比较,IL-12明显增高(P均<0.05);轻型和重型在四期的比较:IL-12,IL-6和TNF均有显著的差异性(P均<0.05)。结论检测IL-12,IL-6,TNF等炎症因子对于流行性出血热的临床疾病严重程度有重要的意义。     

Behavioral, Structural and Molecular Changes following Long-term Hippocampal IL-1β Overexpression in Transgenic Mice

Chronic neuroinflammation is associated with many neurodegenerative and neurocognitive disorders, yet few animal models exist to study the behavioral effects of prolonged neuroinflammation. Therefore, we recently developed a transgenic mouse model harboring an interleukin-1β excisional activation transgene (IL-1β^XAT). These mice display localized IL-1β overexpression and resultant neuroinflammation for up to 1 year following transgene induction. Initial behavioral studies demonstrated long-term memory deficits after 2 weeks of hippocampal IL-1β overexpression. In the present studies, we extend these behavioral studies both in scope and timing. We find long-term contextual but not auditory fear memory impairments following 3 months of IL-1β overexpression. On a battery of other behavioral tests, IL-1β overexpression in IL-1β^XAT mice increased locomotor activity, especially in female mice, and had slight anxiolytic effects. No differences were found in operant conditioning or in basal or stress-induced CORT levels, despite profound hippocampal glial activation. Interestingly, the volume of discrete hippocampal cell layers was reduced after 6 but not 3 months of IL-1β overexpression. Therefore, this animal model provides a novel tool for examining the effects of chronic inflammation on discrete brain regions.     

氯丙嗪、地塞米松对健康志愿者分泌TNF-α,IL-1β,IL-1受体拮抗剂和IL-10的影响

肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)是两种重要的初始炎性细胞因子,能够促进感染性休克的发生和发展。而白介素-1受体拮抗剂(IL-lra)和白介素-10(IL-10)是两种抗炎细胞因子,能对抗前两种因子的作用。氯丙嗪是一种吩噻嗪类衍生物,动物实验表明它能够减少血浆中TNF-α和IL-1β水平,增加IL-10水平,并且对内毒素血症鼠具有保护作用。本研究的目的是观察氯丙嗪对调节人类上述几种因子有无作用,探索其临床实用价值。试验方法:(1)健康志愿者18人,随机分为3组,即氯丙嗪组、地塞米松组、安慰剂组,于用药前后采血。血标本一组立即离心…     

Hexavalent chromium induced ROS formation,Akt, NF-κB,and MAPK activation,and TNF-α and IL-1α production in keratinocytes

In certain cell types, it has been found that, hexavalent chromium could increase ROS formation, activate cell signaling and stimulate the release of cytokines. But, in keratinocytes, these effects have not yet fully been demonstrated. Our aim is to observe the above effects of hexavalent chromium on keratinocytes. By utilizing HaCaT cells and the skin of albino guinea pigs, we showed that hexavalent chromium could increase ROS formation, activate the Akt, NF-kB, and MAPK pathways as well as increase the production of cytokines, including TNF-α and IL-1α. The release of these cytokines from keratinocytes is considered to be a key participant in the pathogenesis of contact hypersensitivity. Among cement workers, chromium hypersensitivity is an important occupational skin disease issue. Therefore, the observations of our study help us better understand the role of hexavalent chromium on the development of chromium hypersensitivity, which might provide clues for clinicians in the development of chemopreventative agents for the prevention of chromium hypersensitivity among cement workers.     

良性前列腺增生组织中CD8,CD20,IL-6和IL-23的表达

目的： 通过检测良性前列腺增生 (benign prostatic hyperplasia, BPH ) 患者手术标本中CD8,CD20,IL-6和IL-23的表达,探讨炎症免疫在BPH病因与进展中的作用。 方法： 收集60例BPH患者的临床资料及手术标本。采用免疫组化技术检测CD8,CD20,IL-6和IL-23在增生前列腺组织中的表达,将检测不同结果病例的临床数据进行对比性统计学分析。结果：CD8表达阳性49例(82%),CD20表达阳性36例(60%),IL-6表达阳性47例(78%),IL-23表达阳性33例(55%)。在检测指标全部强阳性病例组中,前列腺体积大于全部阴性的病例组,IPSS评分也更高（P<0.01）。 结论：BPH组织中存在着炎症细胞标志物和炎性因子的表达,并且与临床数据密切相关,提示炎症免疫可能在BPH的病因与进展中扮演了重要角色。     

Effect of Liposomal and Free Bisphosphonates on the IL-1β, IL-6 and TNFα Secretion from RAW 264 Cells in Vitro

Purpose. In order to evaluate the possible antiinflammatory action of bisphosphonates, the effect of the drugs on the secretion of proinflammatory cytokines (IL-l, IL-6 and TNF) from macrophages was studied. Liposomes or high concentration of extracellular calcium was used to enhance the intracellular delivery of bisphosphonates. Methods. RAW 264 cells were used as macrophage model, and they were induced with lipopolysaccharide to produce the cytokines. The cytokine concentrations in the culture supernatants were measured with time-resolved fluoroimmunoassay. Results. As a free drug, clodronate and pamidronate, but not etidronate, inhibited LPS-stimulated secretion of the cytokines from macrophage-like RAW 264 cells. Low concentrations of pamidronate, however, induced the IL-6 secretion, and the cytokine inhibitory action at the higher concentrations of pamidronate was attributed to cytotoxicity of the compound. The cytokine induction or toxic effects were not observed with clodronate or etidronate. When the drugs were encapsulated in negatively charged unilamellar liposomes, the inhibitory potency of both clodronate and etidronate enhanced by a factor of 10-20, while that of pamidronate was not increased. The complex formation of bisphosphonates with extracellular calcium, although enhancing the uptake of the compounds by macrophages, did not considerably increase their cytokine inhibitory potency. Conclusions. Bisphosphonates have inhibitory action on cytokine secretion by macrophages. The non-cytotoxic cytokine inhibition by liposome encapsulated clodronate could be beneficial in local inflammatory diseases, where the inflammation is sustained by the excessive amounts of inflammatory cytokines produced by activated macrophages.     

Theobromine mitigates IL-1β-induced oxidative stress,inflammatory response,and degradation of type II collagen in human chondrocytes

Osteoarthritis is one of the major causes of disability in elderly adults. Chondrocytes are responsible for the formation and remodeling of articular cartilage in joint tissue. The dysfunction of chondrocytes is a significant factor in the development of osteoarthritis. In the current study, we found that theobromine, a constituent of the cacao plant, possesses a preventive effect against interleukin (IL)-1β-induced chondrocyte dysfunction. Theobromine ameliorates IL-1β-induced production of cellular reactive oxygen species (ROS) and inflammatory mediators including cyclooxygenase-2 (COX-2) and prostaglandin E₂ (PGE₂). The presence of theobromine suppresses IL-1β-induced inducible nitro oxide synthase (iNOS) expression and cellular nitro oxide (NO) production. Theobromine also suppresses IL-1β-induced production of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1), as well as matrix metalloproteinases (MMP)-3 and MMP-13. Additionally, theobromine mitigates IL-1β-induced type II collagen degradation. Mechanistically, we show that theobromine inhibits IL-1β-induced IκBα activation, nuclear factor-κB (NF-κB) protein p65 accumulation, and transfected NF-κB promoter activity, indicating that theobromine suppresses the NF-κB pathway in chondrocytes. Collectively, our study demonstrates that the natural molecule theobromine has a protective effect to counter cytokine-induced chondrocyte dysfunction, implying its beneficial effect in the prevention of osteoarthritis.