Intrathecal bone marrow stromal cells inhibit neuropathic pain via TGF-β secretion

Neuropathic pain remains a pressing clinical problem. Here, we demonstrate that a local, intrathecal (i.t.) injection of bone marrow stromal cells (BMSCs) following lumbar puncture alleviates early- and late-phase neuropathic pain symptoms, such as allodynia and hyperalgesia, for several weeks in murine chronic constriction injury (CCI) and spared nerve injury models. Moreover, i.t. BMSCs reduced CCI-induced spontaneous pain and axonal injury of dorsal root ganglion (DRG) neurons and inhibited CCI-evoked neuroinflammation in DRGs and spinal cord tissues. BMSCs secreted TGF-β1 into the cerebrospinal fluid, and neutralization of TGF-β1, but not IL-10, reversed the analgesic effect of BMSCs. Conversely, i.t. administration of TGF-β1 potently inhibited neuropathic pain. TGF-β1 acted as a powerful neuromodulator and rapidly (within minutes) suppressed CCI-evoked spinal synaptic plasticity and DRG neuronal hyperexcitability via TGF-β receptor 1–mediated noncanonical signaling. Finally, nerve injury upregulated CXCL12 in lumbar L4–L6 DRGs, and this upregulation caused migration of i.t.-injected BMSCs to DRGs through the CXCL12 receptor CXCR4, which was expressed on BMSCs. BMSCs that migrated from the injection site survived at the border of DRGs for more than 2 months. Our findings support a paracrine mechanism by which i.t. BMSCs target CXCL12-producing DRGs to elicit neuroprotection and sustained neuropathic pain relief via TGF-β1 secretion.     

TNF-α contributes to spinal cord synaptic plasticity and inflammatory pain: distinct role of TNF receptor subtypes 1 and 2

Tumor necrosis factor-alpha (TNF-α) is a key proinflammatory cytokine. It is generally believed that TNF-α exerts its effects primarily via TNF receptor subtype-1 (TNFR1). We investigated the distinct roles of TNFR1 and TNFR2 in spinal cord synaptic transmission and inflammatory pain. Compared to wild-type (WT) mice, TNFR1- and TNFR2-knockout (KO) mice exhibited normal heat sensitivity and unaltered excitatory synaptic transmission in the spinal cord, as revealed by spontaneous excitatory postsynaptic currents in lamina II neurons of spinal cord slices. However, heat hyperalgesia after intrathecal TNF-α and the second-phase spontaneous pain in the formalin test were reduced in both TNFR1- and TNFR2-KO mice. In particular, heat hyperalgesia after intraplantar injection of complete Freund’s adjuvant (CFA) was decreased in the early phase in TNFR2-KO mice but reduced in both the early and later phase in TNFR1-KO mice. Consistently, CFA elicited a transient increase of TNFR2 mRNA levels in the spinal cord on day 1. Notably, TNF-α evoked a drastic increase in spontaneous excitatory postsynaptic current frequency in lamina II neurons, which was abolished in TNFR1-KO mice and reduced in TNFR2-KO mice. TNF-α also increased N-methyl-d-aspartate (NMDA) currents in lamina II neurons, and this increase was abolished in TNFR1-KO mice but retained in TNFR2-KO mice. Finally, intrathecal injection of the NMDA receptor antagonist MK-801 prevented heat hyperalgesia elicited by intrathecal TNF-α. Our findings support a central role of TNF-α in regulating synaptic plasticity (central sensitization) and inflammatory pain via both TNFR1 and TNFR2. Our data also uncover a unique role of TNFR2 in mediating early-phase inflammatory pain.     

Effect of triptolide on secretion of inflammatory cellular factors TNF-α and IL-8 in peritoneal macrophages of mice activated by lipopolysaccharide

BACKGROUND:

Research has been carried out to look for safe and effective anti-inflammation drugs from traditional Chinese herbal medicine. As a powerful research technology of life science, molecular biology has entered many areas of traditional Chinese medicine. This study aimed to investigate the effect of triptolide on tumor necrosis factor-a (TNF-α) and interleukin-8 (IL-8) of peritoneal macrophages activated by lipopolysaccharide (LPS) in mice.

METHODS:

Peritoneal elicited macrophages were separated, purified and activated by LPS in mice, then cultured in vitro with triptolide at different concentrations. The activity of TNF-α and the level of IL-8 of cellular supernatants were determined by MTT colorimetric assay and ELISA, respectively.

RESULTS:

The activity of TNF-α in macrophages was significantly inhibited (P<0.01) by triptolide (10^-1-10¹μg/ml) during 4-24 hours in a time- and dose-dependent manner. The level of IL-8 in macrophages was significantly inhibited (P<0.01) by triptolide (10^-1-10¹μg/ml) in 12 hours in a dose-dependent manner.

CONCLUSION:

Triptolide could inhibit the activity of TNF-α and the level of IL-8 in macrophages activated by LPS.KEY WORDS: Triptolide, Lipopolysaccharide, Macrophage, Tumor necrosis factor-α, Interleukin-8     

TRAF6 upregulation in spinal astrocytes maintains neuropathic pain by integrating TNF-α and IL-1β signaling

The proinflammatory cytokines tumor necrosis factor (TNF) α and interleukin (IL) 1β have been strongly implicated in the pathogenesis of neuropathic pain, but the intracellular signaling of these cytokines in glial cells is not fully understood. TNF receptor-associated factor 6 (TRAF6) plays a key role in signal transduction in the TNF receptor superfamily and the IL-1 receptor superfamily. In this study, we investigated the role of TRAF6 in neuropathic pain in mice after spinal nerve ligation (SNL). SNL induced persistent TRAF6 upregulation in the spinal cord. Interestingly, TRAF6 was mainly colocalized with the astrocytic marker glial fibrillary acidic protein on SNL day 10 and partially expressed in microglia on SNL day 3. In cultured astrocytes, TRAF6 was upregulated after exposure to TNF-α or IL-1β. TNF-α or IL-1β also increased CCL2 expression, which was suppressed by both siRNA and shRNA targeting TRAF6. TRAF6 siRNA treatment also inhibited the phosphorylation of c-Jun N-terminal kinase (JNK) in astrocytes induced by TNF-α or IL-1β. JNK inhibitor D-NKI-1 dose-dependently decreased IL-1β–induced CCL2 expression. Moreover, spinal injection of TRAF6 siRNA decreased intrathecal TNF-α– or IL-1β–induced allodynia and hyperalgesia. Spinal TRAF6 inhibition via TRAF6 siRNA, shRNA lentivirus, or antisense oligodeoxynucleotides partially reversed SNL-induced neuropathic pain and spinal CCL2 expression. Finally, intrathecal injection of TNF-α–activated astrocytes induced mechanical allodynia, which was attenuated by pretreatment of astrocytes with TRAF6 siRNA. Taken together, the results suggest that TRAF6, upregulated in spinal cord astrocytes in the late phase after nerve injury, maintains neuropathic pain by integrating TNF-α and IL-1β signaling and activating the JNK/CCL2 pathway in astrocytes.     

Adalimumab – a new TNF-α antibody for treatment of inflammatory joint disease

Tumour necrosis factor alpha (TNF-α) is a pro-inflammatory cytokine with various roles in inflammatory processes. Several TNF blockers are currently approved for use in rheumatoid arthritis (RA) as well as in other inflammatory arthropathies. The latest of these compounds is the human monoclonal antibody, adalimumab, which was obtained using phage display technology and successfully produced in a mammalian expression system. Clinical application of this compound led to significant improvement in patients suffering from RA, both as monotherapy and in combination with various disease modifying antirheumatic drugs (DMARDs), including methotrexate (MTX). Moreover, radiographic progression is significantly inhibited and quality of life improved. This article summarises the available information.     

Macrophage inflammatory protein-1α mediates the development of neuropathic pain following peripheral nerve injury through interleukin-1β up-regulation

In the present study, we investigated the role of the macrophage inflammatory protein-1α (MIP-1α) in the pathogenesis of neuropathic pain following partial sciatic nerve ligation (PSL) in mice. MIP-1α mRNA and its protein were dramatically up-regulated after PSL, and MIP-1α was localized on macrophages and Schwann cells in the injured sciatic nerve (SCN). PSL-induced long-lasting tactile allodynia and thermal hyperalgesia were prevented by the perineural injection of anti-MIP-1α (2 ng). Intraneural (20 ng) and perineural (100 ng) injection of recombinant MIP-1α elicited tactile allodynia and thermal hyperalgesia in sham-operated limb. MIP-1α receptors (CCR1 and CCR5) mRNA and their proteins were also up-regulated in the SCN after PSL, and were localized on macrophages and Schwann cells. PSL-induced tactile allodynia was attenuated by perineural injection (0.2 nmol) of siRNA against CCR1 and CCR5. On the other hand, PSL-induced thermal hyperalgesia was prevented by siRNA against CCR5, but not CCR1. Interleukin-1β (IL-1β) mRNA and its precursor protein in macrophages and Schwann cells were also up-regulated in the SCN after PSL, and PSL-induced neuropathic pain was prevented by the perineural injection of anti-IL-1β (2 ng). PSL-induced IL-1β up-regulation was suppressed by anti-MIP-1α and siRNA against CCR1 and CCR5. Perineural injection of nicotine (20 nmol), a macrophage suppressor, prevented PSL-induced neuropathic pain and suppressed MIP-1α and IL-1β expressions. In conclusion, we propose a novel critical molecule MIP-1α derived from macrophages and Schwann cells that appears to play a crucial role in the development of neuropathic pain induced by PSL.     

前列腺素E1对ARDS病人TNF-α IL-1β IL-6 IL-8的影响

目的 观察PGE1对急性呼吸窘迫综合征(RDS)病人细胞因子TNF-α、IL-1β、IL-6、IL-8的影响,探讨PGE1治疗ARDS的可能作用机制。方法 61例ARDS病人随机分为PGE1治疗组和对照组,应用放射免疫技术测定ARDS机械通气即刻、48h、5d时细胞因子TNF-α、IL-1β、IL-6、IL-8的水平。结果 PGE1治疗组在ARDS机械通气48h、5d时细胞因子TNF-α、IL-1β、IL-6、IL-8活性比对照组显著降低(P<0.01)。结论 PGE1可降低ARDS病程细胞因子活性。     

血糖波动与危重患者CRP TNF-α IL-6水平关系及预后的相关性研究

目的 探讨血糖波动与危重患者血清C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平关系及和预后的相关性.方法 选择ICU危重患者60例.①按APACHEⅡ评分分为三组.采用动态血糖监测系统(CGMS)监测患者入院72 h内平均血糖波动幅度(MAGE);测定空腹血清CRP、TNF-α和IL-6水平;观察28 d患者的预后情况.②回顾分析:根据危重患者入科第1个24 h测得MAGE值按由小到大排序,分四组,比较MAGE与APACHEⅡ评分和病死率的关系.结果 ①三组患者入科后MAGE与血清CRP、TNF-α、IL-6水平明显升高,与正常对照组比较差异有统计学意义(P＜0.01),且MAGE与血清CRP、TNF-α及IL-6水平呈正相关(P＜0.01).②经回顾分析提示,入科MAGE最高的第Ⅳ组患者,APACHEⅡ评分及病死率也最高(P＜0.01),并且入科MAGE与APACHEⅡ评分呈正相关(P＜0.01).结论 危重患者MAGE与血清炎症因子水平密切相关,且与患者危重程度一致,对判断预后具备一定意义.     

急性心肌梗死患者急诊介入前后冠状窦血TNF-α IL-6及CRP水平的动态变化

目的 检测急性心肌梗死(AMI)患者急诊介入治疗前后冠状窦局部肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及C-反应蛋白(CRP)水平的动态变化,并探讨其临床意义.方法 选取入院3～5 h已确诊的AMI患者96例,其中84例有条件置入支架实现再通(再通组),12例因三支病变严重行保守治疗(未通组).再通组分别于冠状动脉开通前及开通后0、10、30、60、120、180 min各时间点抽取冠状窦血,应用ELISA法检测血清中TNF-α和IL-6的水平,以散射比浊法定量测定血清CRP的水平.结果 再通组冠状窦内TNF-α和CRP水平在开通初期呈升高趋势,随着血管再通其水平逐渐下降,并分别于开通后30 min出现波峰;在开通后120 min和180 min时,再通组冠状窦内TNF-α和CRP水平较未通组明显降低;在开通后各时间点,再通组IL-6水平与未通组比较差异均无统计学意义,两组均呈上升趋势.结论 与外周静脉血比较,冠状窦内TNF-α、IL-6及CRP的检测能更准确、更及时地反映局部急性炎症反应的变化,且介入治疗使梗阻血管再通后,能明显改善局部急性炎症反应,但再灌注和支架会对梗阻血管造成一定损伤.     

Amlodipine and atorvastatin improve ventricular hypertrophy and diastolic function via inhibiting TNF-α, IL-1β and NF-κB inflammatory cytokine networks in elderly spontaneously hypertensive rats

This study aimed to examine the effects of amlodipine and atorvastatin alone or in combination on the regulation of inflammatory cytokines and the underlying mechanisms in elderly spontaneously hypertensive (SH) rats. The level of serum hs-CRP was detected with ELISA. The serum TNF-α and IL-1β levels were assessed by radioimmunity assay (RIA). Cardiac inflammatory cell infiltration was observed by HE staining. The protein levels of TNF-α, IL-1β, of NF-κB P65 and IκBα were detected by immunoblotting. The intracellular localization of NF-κB p65 was observed using immunohistochemistry. Amlodipine or atorvastatin obviously ameliorated the myocardial inflammatory cell infiltration in SH rats, which was further improved by combinatorial treatment with amlodipine and atorvastatin. Either amlodipine or atorvastatin decreased plasma IL-1β content in SH rats, but there was no significant difference when compared with untreated SH rats. However, the combination of amlodipine and atorvastatin significantly decreased plasma IL-1β level in SH rats. Moreover, amlodipine or atorvastatin intervention significantly reduced myocardial TNF-α and IL-1β protein levels in SH rats, which was further suppressed by the combination of amlodipine and atorvastatin. In addition, amlodipine or atorvastatin inhibited the activity of NF-κB signaling in SH rats, which was further suppressed by combinatorial treatment. Furthermore, amlodipine or atorvastatin restored the activity of IκB-α in SH rats, which was enhanced by combinatorial treatment. Our results demonstrated amlodipine and atorvastatin improved ventricular hypertrophy and diastolic function possibly through the intervention of TNF-α, IL-1β, NF-κB/IκB inflammatory cytokine network. Our study suggests that amlodipine combined with atorvastatin may have additive effect on inhibiting inflammatory response.     

Changes in Glucose,TNF-α and IL-6 Blood Levels in Middle-aged Women Associated with Aerobic Exercise and Meditation Training

[Purpose] The purpose of this study was to investigate the effects of exercise therapy on glucose, TNF-α and IL-6 blood levels in middle-aged women. [Subjects] A total of 46 participants were assigned to four groups: Type D personality+Exercise (n=12), Type D+no-Exercise (n=12), not-Type D+Exercise (n=12), and not-Type D+no-Exercise (n=10). [Methods] Blood glucose was measured by the hexokinase method. An enzyme-linked immunosorbent assay (ELISA) was used to measure the circulating plasma levels of TNF-α and IL-6 (Quantikine HS, R&D Systems, Minneapolis, USA). An aerobic exercise program and meditation were conducted in parallel by the Exercise groups for 10 months. Stretching was performed for 10 min as a warm-up, and then walking and running on a treadmill at 60 to 70% of HRmax were performed for 40 min three times a week. Blood samples were processed according to standard laboratory procedures. [Results] Fasting glucose showed a significant interaction effect among groups, group×time, and post-test. TNF-α showed a significant difference among groups, and was lower in the not-Type D personality+Exercise group than in the other three groups. IL-6 showed a significant difference among the groups. [Conclusion] In conclusion, personality may affect the established effect of exercise on cytokine activity. Type D personality was independently associated with significant immune activation, and increase in TNF-α activity was observed among Type D participants.Key words: Glucose, TNF-α, IL-6 blood levels     

TNF-α −308G > A and IL-6 −174G > C polymorphisms in Tunisian patients with coronary artery disease

ObjectiveOur aim was to evaluate the contribution of tumor necrosis factor (TNF)-α ?308G > A and interleukin (IL)-6 ?174G > C gene promoter variants to the presence of coronary artery disease (CAD) in Tunisians.Design and methodsStudy subjects comprised 418 angiographically proven CAD patients and 406 age-, gender-, and ethnic origin-matched controls. Genotyping was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis.ResultsThere were no significant differences in the allelic distribution of TNF-α ?308A (19.6% vs. 19.0%, P = 0.73), and IL-6 ?174C (15.6% vs. 14.3%, P = 0.47) promoter polymorphisms between CAD patients and control subjects, respectively. In addition, single locus analysis revealed no differences in genotype frequencies between the two study groups, and the combined distribution of both genotypes did not differ significantly between controls and CAD patients (P > 0.05).ConclusionThere is no allelic or genotypic association of TNF-α ?308G > A and IL-6 ?174G > C promoter polymorphisms with CAD in Tunisians, thereby confirming an ethnic-selective contribution of both gene variants to CAD presence.     

肺咳颗粒联合抗生素治疗风热袭肺夹滞型肺部感染患儿的疗效及对血清IL-6 TNF-α INF-γ水平的影响

目的 探讨肺咳颗粒联合抗生素治疗风热袭肺夹滞型肺部感染患儿的临床疗效及对血清白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(INF-γ)水平的影响。方法 将108例风热袭肺夹滞型肺部感染患儿按随机数字表法分为研究组和对照组,各54例。对照组患儿静脉滴注阿奇霉素治疗,研究组患儿在对照组基础上口服肺咳颗粒治疗,两组患儿均治疗1周。观察临床疗效,比较两组患儿治疗前后中医症候相关评分、肺功能以及血清IL-6、TNF-α、INF-γ水平变化。结果 研究组患儿治疗总有效率为98.15%,对照组为85.19%,研究组高于对照组（P<0.05）。治疗后,两组患儿中医症候积分均较治疗前降低,且研究组低于对照组（P<0.01）;两组患儿潮气量、用力呼气25%流速均较治疗前升高,且研究组高于对照组（P<0.01）;两组患儿血清IL-6、TNF-α、INF-γ水平均较治疗前降低,且研究组低于对照组（P<0.01）。两组不良反应发生率比较,差异无统计学意义（P>0.05）。结论 肺咳颗粒联合抗生素治疗儿童风热袭肺夹滞型肺部感染的临床疗效显著,可有效控制患儿症候,...