Dorsal root sensitivity to interleukin-1 beta, interleukin-6 and tumor necrosis factor in rats

The release of inflammatory cytokines caused by a disrupted disc may play a critical role in pain production at nerve endings, axons, and nerve cell bodies. Herniated disc tissue has been shown to release inflammatory cytokines such as interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor (TNF), and other algesic chemicals. This study was designed to characterize the effects of these proinflammatory cytokines on the somatosensory neural response at the dorsal root level in rats. It is hypothesized that their effects on nerve endings in disc and adjacent tissue contribute to low-back pain, and the effects on dorsal root axons and ganglia contribute to radiculopathy and sciatica. Surgically isolated sacral dorsal roots were investigated by electrophysiologic techniques. IL-1beta, IL-6, or TNF (100 ng, each) were applied onto the dorsal roots. Neural responses and mechanosensitivity of the receptive fields were evaluated over time. The results showed that 3 h after each cytokine application, the neural activity was statistically decreased. The mechanical sensitivity of the receptive fields increased at 90 min following IL-1beta or TNF application, and returned to normal more than 3 h after IL-1beta application. IL-1beta, IL-6, and TNF may be neurotoxic to dorsal root axons. Furthermore IL-1beta and TNF may sensitize the peripheral receptive fields. This study suggests that dorsal roots may be impaired by these proinflammatory cytokines.     

终末糖基化产物对神经小胶质细胞IL-1β、IL-6表达的影响

目的：研究终末糖基化产物（AGEs）对神经小胶质细胞白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）表达的影响。方法：以不同浓度、不同时间AGEs刺激体外培养的小鼠小胶质瘤细胞（BV-2细胞）,观察细胞形态的变化,应用逆转录聚合酶链反应（RT-PCR）检测细胞内IL-1β、IL-6mRNA水平,应用酶联免疫吸附法（ELISA）测定细胞培养上清液中IL-1β、IL-6的蛋白含量。结果：与空白组和AGEs对照物组比较,AGEs组细胞活化呈阿米巴样,胞核大而圆,核仁明显,胞浆内颗粒物增多,以300mg/L作用18h组效果最显著。与空白组和AGEs对照物组比较,作用时间相同时,AGEs浓度为50mg/L时即可见细胞的IL-1β、IL-6mRNA水平明显增加（P〈0.05或P〈0.01）,浓度为150mg/L时,细胞培养上清液中IL-1β、IL-6的蛋白含量明显增加（P〈0.05）;作用浓度相同时,2h即可见细胞的IL-1βmRNA水平明显增加（P〈0.05）,6h见细胞的IL-6mRNA水平和细胞培养上清液中IL-1β、IL-6的蛋白含量明显增加（P〈0.05或P〈0.01）,其中以300mg/L作用18h组效果最显著（P〈0.05）。结论：AGEs刺激可使小胶质细胞内IL-1β、IL-6高表达。     

Cytokines in stored red blood cell concentrates: promoters of systemic inflammation and simulators of acute transfusion reactions?

Background: The cytokine network has important implications for the systemic inflammatory and metabolic response in trauma and infection. Cytokines exogenously administered to traumatized and infected patients may have implications for the trauma response in these patients. The main objective of this study was to characterize red blood cell concentrates (RBCs) with regard to cytokine content.
Methods: We investigated the concentrations of tumor necrosis factor α (TNF), interleukin-1 β (IL-1), interleukin-6 (IL-6) and interleukin-8 (IL-8) in sixteen units of RBCs stored at +4° C during 40 days. Samples from RBCs were taken every tenth day. Healthy volunteers were used as controls.
Results: IL-1 and IL-8 in RBCs were increased compared to controls, P< 0.01 - P < 0.001 and TNF in RBCs were increased on days 1 and 40 compared to controls, P < 0.05. During storage TNF was highest on day 1, 69 (< 3 - 1060) pg/ml, median (range). IL-1 concentrations increased during the period of storage from 5 (< 2 - 205) pg/ml to 174 (< 2 - 2180) pg/ml, P < 0.01. IL-6 was 6 (< 2 - 210) pg/ml on day 1 and did not change over the period of storage. IL-8 was highest on day 40,164 (15 - 790) pg/ml and compared to day 1 the concentrations were increased on day 10 and day 40, P < 0.05 for both comparisons.
Conclusions: The results indicate the presence of TNF, IL-1, IL-6 and IL-8 in stored RBCs, though there was a great variability over the period of storage and between units of RBCs. In some samples of RBCs the content of cytokines reached levels that may be anticipated to contribute to systemic inflammation and the symptomatology of acute transfusion reactions.     

大鼠烧伤后库普弗细胞在促炎细胞因子产生中的作用

目的　观察大鼠严重烧伤后早期 ,库普弗细胞在肿瘤坏死因子α(TNFα)、白细胞介素(IL) 1β、IL 6产生中的作用。方法　观察 (1)烧伤血清对体外培养的大鼠库普弗细胞分泌TNFα、IL 1β、IL 6的刺激作用 ;(2 )烧伤后大鼠库普弗细胞的细胞因子mRNA表达变化 ;(3)应用库普弗细胞特异性抑制剂三氯化钆后 ,烧伤大鼠血浆内细胞因子含量变化。　结果　烧伤血清能刺激库普弗细胞释放TNFα、IL 1β、IL 6 ;大鼠烧伤后库普弗细胞TNFα、IL 1β、IL 6mRNA表达量显著升高 ;预先抑制库普弗细胞的活性 ,烧伤后血浆TNFα、IL 1β、IL 6水平均显著降低 ,分别为烧伤组的 34.71%、36 99%、33.70 %。结论　库普弗细胞是大鼠烧伤后血浆中TNFα、IL 1β、IL 6的主要来源     

Effect of interleukin 6 on basal and FSH-stimulated secretion of estradiol and progesterone by human granulosa cells in vitro

Aim: To determine the effects of interleukin-6 (IL-6) on the secretion ofestradiol and progesterone by human granulosa cells in vitro. Methods:Granulosa cells were obtained from infertile patients undergoing IVF ETtreatment and cultured with serum-free supplemented HAM's F10 medium.In the absence or presence of FSH, granulosa cells were treated with differ-ent concentrations of gene recombinant human iterlukin-6 (rhIL-6). Themedia were collected after 24, 48, 72 and 96 h and assayed for estradiol andprogesterone. IL-6 and R mRNA was determined by means of RNA slotblot. Results: IL-6 had a significant inhibitory effect on estradiol secre-tion, especially in the presence of FSH. IL-6 inhibited the FSH-stimulatedprogesterone secretion, but not the basal progesterone release. The inhibi-tion shows certain degrees of dose- and time-dependency. Conclusion:IL-6 participates in the regulation of ovarian function through its inhibitoryeffect on FSH-stimulated steroidogenesis by granulosa cells.     

几种抗炎药对大鼠抗炎及血清细胞分子影响的研究

目的比较研究非甾体抗炎药醋氯芬酸、双氯芬酸和布洛芬对佐剂性关节炎大鼠的抗炎作用,并探讨其作用机制。方法应用弗氏完全佐剂制作AA动物模型,将Wistar大鼠35只随机分为正常对照组、模型组、醋氯芬酸组、双氯芬酸组和布洛芬组5组,通过组织学及双抗体夹心ELISA法检测比较5组大鼠关节局部的炎症情况,及其外周血细胞因子IL-1β和IL-6的含量。结果在醋氯芬酸组、双氯芬酸组和布洛芬组大鼠关节炎的炎症反应明显轻于模型组。细胞因子IL-1β和IL-6的含量在醋氯芬酸组、双氯芬酸组和布洛芬组均低于模型组（P〉0.05）;且醋氯芬酸组低于双氯芬酸组和布洛芬组（P〉0.05）;而双氯芬酸组和布洛芬组间无显著差异（P〈0.05）。结论醋氯芬酸、双氯芬酸和布洛芬均能降低AA大鼠血中的IL-1β和IL-6含量,有较好的抗炎作用,醋氯芬酸作用相对较为显著。     

Effects of interleukin-1 beta, interleukin-6, and tumor necrosis factor on sensitivity of dorsal root ganglion and peripheral receptive fields in rats

This study was designed to characterize the effects of low doses (0.5–5 ng) of pro-inflammatory cytokines, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF), on the neural activity of dorsal root ganglion (DRG) in rats. The purpose of this study was to examine the effects of cytokines (IL-1β, IL-6, and TNF) on the somatosensory neural response of DRG. The release of inflammatory cytokines by an injured disc may play a critical role in pain production at nerve endings, axons, and nerve cell bodies. Herniated disc tissue has been shown to release IL-1β, IL-6, TNF, and other algesic chemicals. Their effects on nerve endings in disc and adjacent tissue may lead to low back pain and their effects on dorsal root axons and ganglia may lead to sciatica. Exposed lumbar DRGs were investigated by electrophysiologic techniques. Sham (mineral oil), control (carrier solution), or IL-1β, IL-6, or TNF at doses of 0.5, 1, and 5 ng were applied over the DRG. Baseline discharge rates as well as mechanosensitivity of the DRG and peripheral receptive fields were evaluated over 30 min. Applications of IL-1β at 1 ng resulted in an increase in the discharge rate, 5 ng resulted in an increased mechanosensitivity of the DRG in group II units. Similarly, after 1 ng TNF applications, group II units also showed an increase in mechanosensitivity of DRG and peripheral receptive fields. At low doses IL-1β and TNF sensitization of receptive fields were observed. The responses observed in the group II units indicate that a sub-population of afferent units might have long-term effects modifying the sensory input to the central nervous system. This study provides added evidence to the role of cytokines in modulating afferent activity following inflammation.     

Interleukin-6 is both necessary and sufficient to produce perioperative neurocognitive disorder in mice

Background

Perioperative neurocognitive disorders (PND) result in long-term morbidity and mortality with no effective interventions available. Because interleukin-6 (IL-6), a pro-inflammatory cytokine, is consistently up-regulated by trauma, including after surgery, we determined whether IL-6 is a putative therapeutic target for PND in a mouse model.

Strong inflammatory cytokine response in male and strong anti-inflammatory response in female kidney transplant recipients with urinary tract infection

Urinary tract infection (UTI) is the most common post-transplant infection in renal transplant recipients. The relationship of plasma and urine cytokines with UTI after kidney transplantation has not yet been delineated and literature reports on cytokine and UTI are rare. In a retrospective study, we compared post-transplant plasma and urine cytokine levels of 132 outpatient renal transplant recipients with or without UTI. Soluble interleukin-1 receptor antagonist (sIL-1RA), IL-2, sIL-2R, IL-3, IL-4, IL-6, sIL-6R, IL-8, IL-10, transforming growth factor-beta2 (TGF-beta2), interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) levels were determined using commercially available enzyme-linked immunosorbent assay (ELISA) kits. We found gender-related urine cytokine patterns. Anti-inflammatory sIL-1RA was significantly higher in females than in males and this gender-related difference was more pronounced in bacteriuric (P < 0.0001) than in nonbacteriuric (P = 0.001) patients. Urine proinflammatory cytokines IL-6 (P = 0.001) and IL-8 (P = 0.007) were significantly higher in male patients with bacteriuria than in males without bacteriuria and sIL-2R (P = 0.001) and sIL-6R (P = 0.03) were significantly higher in males with leukocyturia than in males without leukocyturia. Bacteriuria in males was associated with higher doses of immunosuppressive drugs (P = 0.02). Male renal transplant recipients with UTI have a strong inflammatory cytokine response with activation of IL-6, IL-8, sIL-2R and sIL-6R producing cells, whereas female patients with UTI block the inflammatory response to UTI by production of sIL-1RA.     

TAKEDA-143242 increased survival via reduced cytokines in porcine peritonitis

TAKEDA-143242 (TAK-242) is a small molecule shown to inhibit lipopolysaccharide-induced intracellular signaling and inflammation. In vitro studies demonstrated that TAK-242 can prevent release of TNF-α, IL-1β, and IL-6 from activated macrophages of several species, including pigs. This study tested the hypothesis that TAK-242 would protect pigs from lethal gram-negative peritonitis via an anti-cytokine mechanism. A validated model of porcine gram-negative peritonitis, which employs chronically inplantated cardiac transducers and aortic and pulmonary artery catheters, was used. Pigs were pretreated with TAK-242 or its vehicle via a blinding procedure prior to intraperitoneal implantation of an LD(90) dose of E. coli 0111:B4 in a fibrin clot. Ten pigs were treated with TAK-242 and nine with its vehicle. All ten TAK-242 treated pigs survived, while three of the nine vehicle treated pigs survived (P = 0.01 χ(2) test). Pulmonary artery pressure increased markedly in vehicle pigs, and this elevation was significantly (two-way ANOVA) obviated in TAK-242 treated group. Circulating levels of cytokines in vehicle treated pigs showed increased expression (3930 ± 1770 at 1 h, 1007 ± 400 TNF-α at 2 h; 719 ± 308 of IL-1β at 2-6 h; 33000 ± 1000 of IL-6 at 2-4 h [pg/mL, mean ± SEM]). Peak circulating levels of these cytokines were significantly reduced by pretreatment with TAK-242 (<25 pg/mL TNF-α ; <100 pg/mL IL-1β; 0-1700 pg/mL IL-6, peak values). This study found that pretreatment with TAK-242 yielded significantly positive survival benefit in a lethal sepsis model that was associated with improved cardiovascular status and suppressed cytokine release.     

The calcium channel blocker nisoldipine minimizes the release of tumor necrosis factor and interleukin-6 following rat liver transplantation

Kupffer cells, when activated, release toxic cytokines such as tumor necrosis factor (TNF), which can cause tissue injury. Takei et al. have reported that nisoldipine, a calcium channel blocker which decreases phagocytotic activity by Kupffer cells, also diminishes liver and lung injury and dramatically improves survival following liver transplantation [27]. Therefore, we studied the effect of nisoldipine on the time course of TNF and interleukin-6 (IL-6) release following cold storage and liver transplantation in the rat. Livers were stored under survival and non-survival conditions in cold Euro-Collins solution in the presence or absence of nisoldipine (1.4 µM). After storage, the effluent was collected for determination of cytokines. The liver was then transplanted orthotopically and serum was collected at various time intervals for up to 5 h. In the effluent, TNF levels were very low in both the control and nisoldipine-treated groups and IL-6 was not measurable. Furthermore, when livers were stored under survival conditions and transplanted (liver stored in the cold for 4 h), serum TNF (2 U/ml) and IL-6 (350 U/ml) values were minimal in both the control and nisoldipine-treated groups. In contrast, when livers were stored under non-survival conditions and transplanted (liver stored in the cold for 10 h), TNF levels increased to 15 ± 2 U/ml, 150 min after graft reperfusion, an increase which was prevented by nisoldipine (6.5 U/ml). Serum IL-6 levels were also elevated 300 min after transplantation in livers stored for 10 h. Nisoldipine also reduced the release of this cytokine. Serum transaminases (SGOT) were elevated to values around 2000 U/l 5 h following transplantation. In the nisoldipine-treated group, values were lower between 60 and 300 min. In the lung, interstitial and alveolar edema and cellular infiltration were detectable 5 h post-operatively and were diminished by nisoldipine. These data confirmed that TNF and IL-6 release were minimal following cold storage and transplantation of livers stored under survival conditions, but were elevated transiently after transplantation under non-survival conditions. Nisoldipine prevented cytokine release, most likely by blocking the activation of Kupffer cells, which may explain how it decreases liver and lung injury very early following liver transplantation.     

Shiga toxin-1 regulation of cytokine production by human glomerular epithelial cells

BACKGROUND/AIMS: Inflammatory cytokines may enhance renal injury in post-diarrheal hemolytic uremic syndrome (Stx HUS) by enhancing the cytotoxic effect of Shiga toxins (Stx). The sources of inflammatory cytokines in Stx HUS are unclear. Since Stx-1 potently inhibits protein synthesis by glomerular epithelial cells (GEC) and increases cytokine release by renal epithelial cells, we examined Stx-1 regulation of cytokine production by human GEC. METHODS: Stx-1 (and cycloheximide (CHX), another protein synthesis inhibitor) cytotoxicity, protein synthesis inhibition, and effect on interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) release and mRNA levels were determined. RESULTS: Stx-1 alone had a modest stimulatory effect on inflammatory cytokine production by GEC that occurred at toxin concentrations ranging from minimal to 50% inhibition of protein synthesis. CHX, at concentrations that produced similar inhibition of protein synthesis, increased IL-1, IL-6, and TNF protein release and mRNA accumulation, but in a different time- and dose-dependent pattern than Stx. Lipopolysaccharide (LPS) did not change IL-1, but stimulated IL-6 and TNF production. LPS and Stx-1 combined stimulated production of all three cytokines to a greater extent than either toxin alone. CONCLUSION: These data indicate that: (1) Stx-1 alone modestly stimulates GEC inflammatory cytokine production; (2) LPS and Stx-1 combined can potently enhance GEC cytokine release, and (3) this action of Stx-1 may relate in part to inhibition of protein synthesis but cannot be fully attributed to this effect.     

不同CO2气腹条件对腹腔IL-1β和IL-6的影响

目的探讨不同CO2气腹条件对腹腔IL-1β和IL-6的影响。方法将56只雌性SD大鼠按随机数字表法分成7组,1组为对照组,另6组为气腹组,设定CO2气腹条件为压力0．67kPa,气流量1．01Mmin,时间分别为1、2和3h（C1h、C2h和C3h组）,同时设立C8p组（压力1．07kPa,气流量1．01Mmin,时间1h）、C2f组（压力0．67kPa,气流量2．0L／min,时间1h）和C3f组（压力0．67kPa．气流量3．0L／min,时间1h）。实验后12h采集腹腔液．ELISA方法检测IL-1β和IL-6浓度。结果CO2气腹引起腹腔炎性反应,随着气腹时间和气流量的增加,C2h、C3h和C3f组腹腔液IL-1β和IL-6浓度升高,并显著高于C1h组（P〈0．05）。随着压力的增加,IL-1β和IL-6浓度虽然升高,但与C1h组差异无统计学意义（P〉0．05）。结论腹腔炎性反应与CO2气腹时间和气流量有关。但在工作压力范围内．炎性反应与压力无关,腹腔镜手术应在工作压力范围内进行．采用较小的气流量并同时缩短手术时间。     

他汀药物治疗对尿毒症患者外周血单个核细胞功能的影响

目的：探讨小剂量他汀治疗对尿毒症患者外周血单个核细胞（PBMC）功能的影响。方法：选择尿毒症患者55例,随机分为他汀治疗组（氟伐他汀胶囊,20mg/d）和对照观察组,采用放射免疫法测定患者血清和PBMC培养上清中白细胞介素-1（IL-1）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）含量。另选择20例健康人作为正常对照组。结果：与正常组比较,尿毒症患者血清炎症细胞因子（IL-1、IL-6、TNF-α）含量显著升高（P〈0.05）,PBMC分泌上述炎症细胞因子的能力也显著增强（P〈0.05）。与尿毒症患者对照组比较,他汀治疗组患者血清炎症细胞因子水平明显下降（P〈0.05）,PBMC基础分泌和血管紧张素Ⅱ刺激分泌炎症细胞因子的能力被显著抑制（P〈0.05）。小剂量他汀治疗对血糖、血脂（总胆固醇、低密度脂蛋白胆固醇）均无影响。结论：小剂量他汀治疗可显著抑制尿毒症患者外周血PBMC的活化。     

Cerebrospinal fluid cytokine levels after surgery with spinal or general anesthesia

BACKGROUND AND OBJECTIVES: Studies show that pain may cause neuroinflammatory changes in the spinal cord. These inflammatory changes could be caused by circulating factors such as plasma cytokines or could be a primary neuroimmune response of the central nervous system following peripheral nerve injury. To identify the possible effects of peripheral trauma and pain on the cytokine environment of the spinal cord, interleukin-6 (IL-6) and interleukin-10 (IL-10) concentrations in plasma and cerebrospinal fluid (CSF) were measured before and after hip replacement surgery. METHODS: The investigation used a prospective, observational design to measure cytokine levels in samples of plasma and CSF by enzyme-linked immunosorbent assay (ELISA). Samples were taken from surgical patients before and after surgery under general anesthesia or spinal anesthesia performed with or without a spinal catheter. Reference samples were also obtained from healthy control subjects. RESULTS: Both plasma and CSF levels of IL-6 increased substantially after major surgery with either general or spinal anesthesia. No significant correlation was observed between plasma IL-6 and CSF IL-6 levels, suggesting a central origin for increased CSF cytokine levels. IL-10 did not change in plasma or CSF after surgery. Plasma and CSF IL-6 and IL-10 cytokine levels were very low or undetectable in healthy controls. CONCLUSIONS: Major orthopedic surgery leads to elevated CSF levels of the proinflammatory cytokine, IL-6. The origin of increased CSF IL-6 may be central because there was no significant correlation with plasma levels.     

The cytokine response in a patient undergoing surgery for an ACTH-producing tumor

The administration of glucocorticoids has been used to inhibit the cytokine production in patients undergoing cardiac surgery during cardiopulmonary bypass. In this case report the cytokine response in a 43-year-old man undergoing operation for an ACTH-producing lung tumor is presented. There had been clinical symptoms for about 18 months and 8 months before operation elevated serum concentrations of ACTH and cortisol were measured. The patient underwent thoracotomy with removal of the tumor and the cytokine response in relation to surgery was measured as plasma concentrations of IL-lα, IL-6, TNFα and TNFβ. With the long-lasting high endogenous glucocorticoid production low or undetectable cytokine levels were expected in this patient. However, a marked and prolonged IL-6 and TNFβ response was measured while IL-1α and TNFα were detectable in only a few samples. The cytokine response in this patient was different from the response normally found in patients undergoing major surgery.