白介素-1受体拮抗剂对双后肢大鼠椎间盘细胞凋亡的影响

目的：探讨白介素-1受体拮抗剂（IL-1Ra）对双后肢大鼠椎间盘细胞凋亡的影响。方法：建立双后肢大鼠模型54只，随机分为试验组、空白对照组和阴性对照组，每组18只．试验组从造模当天即予腹腔注射IL-1Ra（50ng／kg体重），隔天1次重复注射至处死，空白对照组不予任何处理，阴性对照组隔天1次予腹腔注射等量生理盐水。于造模后1、3、6个月每组分别处死6只大鼠，完整取出L4／5椎间盘后进行固定、切片，使用末端脱氧核件酸转移酶介导的dUTP缺口末端标记法（TUNEL法）检测各组大鼠椎间盘细胞的凋亡情况，并使用透射电镜观察凋亡细胞的超微结构。取L5/6椎间盘使用流式细胞仪检测各组大鼠椎间盘细胞的凋亡情况。结果：术后1个月试验组、空白对照组及阴性对照组椎间盘细胞的凋亡无显著性差异（P〉0．05）：术后3个月试验组与空白对照组及阴性对照组相比细胞凋亡明显减少，有显著性差异（P〈0．05）；术后6个月试验组与空白对照组及阴性对照组相比细胞凋亡率有非常显著性差异（P〈0．01）。结论：IL-1Ra对双后肢大鼠椎间盘细胞凋亡有明显的抑制作用。     

白介素-1受体拮抗剂(IL-1Ra)与椎间盘基质代谢

临床上因椎间盘退变而引起腰腿痛患病率高达60％～80％，而由于腰椎间盘突出引起坐骨神经痛的患者占总人口数的5％。腰椎间盘突出症已成为临床常见病、多发病，给患者带来巨大的痛苦，同时也消耗了大量的医疗资源和费用。虽然目前对椎间盘退变的确切机理尚不清楚，但随着近年来对椎间盘的分子生物学、免疫学及生物力学等认识的进展，以及实验设备的改善和实验手段的改进，对椎间盘退变机理的研究正在不断深入，特别是对椎间盘基质的代谢及其调节的认识取得长足的进步。本文就椎间盘基质的组成及退变时成分的改变和IL-1Ra对退变椎间盘基质代谢的调节做相关总结。     

白介素-1受体拮抗剂对椎间盘纤维环胶原纤维代谢的影响

目的 探讨白介素-1受体拮抗剂(IL-1Ra)对双后肢大鼠椎间盘纤维环胶原代谢的影响.方法 建立双后肢大鼠模型36只,随机分为试验组和对照组,每组18只.试验组从造模当天即予腹腔注射IL-1Ra(50ng/kg体重),隔天重复注射至处死;对照组18只不予任何处理.于造模后1、3、6个月每组分别处死6只,完整取出L3～4椎间盘固定、切片,予SABC法进行免疫组化,使用图像分析系统对髓核中Ⅰ、Ⅱ型胶原染色进行面积扫描,并使用扫描电镜观察纤维环胶原纤维超微结构.结果 术后第1个月试验组与对照组椎间盘纤维环中Ⅰ、Ⅱ型胶原面积差异无显著性(P＞0.05),第3个月及第6个月试验组与对照组Ⅰ、Ⅱ型胶原面积相比差异有显著性(P＜0.01).结论 IL-1Ra对双后肢大鼠椎间盘纤维环中Ⅰ、Ⅱ型胶原的代谢有重要的作用.     

白介素-1受体拮抗剂对大鼠急性脊髓损伤后神经细胞凋亡和caspase-3表达的影响

目的：观察白介素-1受体拮抗剂（IL—1Ra）对脊髓损伤（SCI）后继发性神经细胞凋亡和caspase-3表达的影响。方法：60只SD大鼠，随机分为假手术组、脊髓损伤组、IL-1Ra治疗组和生理盐水对照组，每组15只。采用Allen’s法建立大鼠急性SCI动物模型．IL—1Ra治疗组和生理盐水对照组于SCI后立即硬膜下腔内分别注射IL-1Ra（10μg／10μl）和等量生理盐水，于伤后24h以损伤为中心（假手术组取相应部位），切取长约8mm脊髓组织．用蛋白印迹法和免疫组化染色法检测caspase-3表达的变化：采用实时定量PCR法检测caspase-3mRNA的表达情况；用原位末端标记法（TUNEL）检测SCI后神经细胞凋亡情况。结果：SCI后24h，SCI组与假手术组比较受损伤的脊髓组织中caspase-3mRNA和蛋白表达水平均显著升高（P〈O．05），且TUNEL阳性细胞明显增多（P〈O．01）：IL—1Ra治疗组大鼠受损伤节段脊髓组织caspase-3表达及TUNEL阳性细胞数较生理盐水对照组）均明显减少（P〈O．01）。结论：IL-1Ra治疗可减少急性SCI后脊髓组织中caspase-3的表达和神经细胞凋亡的发生。     

被动吸烟大鼠椎间盘中白介素-1β和白介素-1受体Ⅰ的表达及意义

目的：观察被动吸烟大鼠椎间盘中自介素-1β（IL-1β）和白介素-1受体Ⅰ（IL-1RⅠ）的表达，探讨被动吸烟诱发椎间盘退变的机制。方法：取4周龄SD大鼠60只，随机分为6组，第1组不予吸烟，10只；第2-4组分别吸烟2周、4周、8周，第5、6组吸烟8周后予停止吸烟4周、8周，每组10只。在相应时间点处死动物，取L4/5椎间盘行HE染色和IL-1β及IL-1RⅠ免疫组化染色，观察椎间盘退变情况和IL-1β及IL-1RⅠ的表达情况。结果：HE染色显示，吸烟2周时椎间盘无明显退变，吸烟4周时椎间盘出现2～3级退变，吸烟8周时椎间盘出现3-4级退变，停止吸烟后4周退变无明显修复；停止吸烟后8周退变部分修复。免疫组化染色显示，吸烟2周组IL-1β及IL-1RⅠ染色阳性细胞率增加，吸烟4周组染色阳性细胞率大于2周组；吸烟8周组达（76±3．2）％和（46±2．8）％，停止吸烟后4周开始下降，8周时降至（66±2．9）％和38±2．2％。结论：被动吸烟可以诱发大鼠椎间盘退变，且随吸烟时间延长退变加重；其导致退变的机制可能与上调椎间盘内IL-1β和IL-1RⅠ的表达有关。     

白细胞介素-1受体拮抗剂对兔椎间盘髓核前列腺素和5-羟色胺代谢的影响

目的:研究白细胞介素-1受体拮抗剂(IL-1Ra)对兔椎间盘髓核前列腺素E2(PGE2)、前列腺素F1琢(PGF1琢)和5-羟色胺(5-HT)代谢的影响。方法:取日本大白兔椎间盘髓核,制成匀浆后进行髓核组织的体外培养,观察不同浓度IL-1Ra和同一浓度IL-1Ra不同培养时间培养液中PGE2、PGF1琢和5-HT的含量。结果:IL-1Ra(100、200、400ng/ml)实验组较对照组培养液中PGE2、PGF1琢和5-HT的含量显著减少(P<0.01),呈现明显的浓度依赖性;IL-1琢(2.0ng/ml)组和IL-1Ra(200ng/ml)组随培养时间延长,PGE2、PGF1琢和5-HT的含量增加,每个时间段实验组与对照组浓度之差的差异有显著性意义(P<0.01)。结论:IL-1Ra可以拮抗IL-1琢对椎间盘髓核细胞PGE2、PGF1琢和5-HT合成的促进作用,呈现明显的浓度依赖性。IL-1Ra可以较长时间拮抗IL-1琢生物学作用,但本实验未能证明此作用具有时间依赖性。     

椎间盘突出症与白细胞介素-1受体拮抗剂

在下腰痛和/或下肢放射性疼痛的病因中，椎间盘突出是常见的病因之一。而腰椎间盘突出的大小并不一定与疼痛的程度成正比[1]。突出的椎间盘组织所致脊神经根受压不是引起神经根痛和功能障碍的唯一因素。来源于腰椎间盘的组织降解产物及其通过免疫或非免疫途径产生的生物活性物质如：细胞因子、前列腺素E2（PGE2）、磷脂酶A2（PLA2）等引起的炎症反应和/或免疫反应使神经根出现炎症损害，导致神经根对机械压迫更敏感，这种观点更令人信服［2～6］。近来大量研究表明，白细胞介素鄄1（IL鄄1）在突出的椎间盘组织中具有很高的活性，且能刺…     

白介素-1受体桔抗剂对实验性新月体肾炎骨调素表达的调节作用

目的探讨白介素-1受体拮抗剂（IL-lra）对实验性新月体肾炎肾脏组织骨调素（OPN）表达的调节作用，进一步阐明IL-1在实验性新月体肾炎发病机制中的作用。方法加速型实验性新月体肾炎模型应用兔抗鼠肾小球基底膜肾毒血清制备。第1组为模型组：注射肾毒血清后第0～7天，不加任何干扰治疗；另外2组为治疗组及治疗对照组：从第7～14天，分别给予IL-lra和生理盐水持续静脉点滴。结果模型组OPN的表达显著增高。生理盐水治疗组，OPN在肾小球和小管-间质细胞的表达更高，并和巨噬细胞的局部浸润及肾功能密切相关。而IL-lra治疗组，OPN的表达显著下调，巨噬细胞浸润明显减少，部分逆转肾功能。结论应用IL-lra抑制IL-1的活性可显著下调骨调素的表达和保护肾功能，提示IL-lra的治疗作用可能是通过抑制骨调素的表达，减少巨噬细胞在肾脏局部浸润的机制。     

老年男性骨密度与白介素-1受体拮抗剂及有关生化指标关系的研究

目的 探讨白介素－1及其受体拮抗剂在老年男性骨代谢中的作用。方法 老年男性组133例，老年前期男性组35例。酶联免疫法（ELISA）测定血浆白介素－1β（IL－1β）、IL－1受体拮抗剂（IL－1Ra)、血清骨钙素（BGP）、尿I型胶原C末端肽（CTX）。放免法叫雌二醇（E2）、总睾酮（T）。结果 与老年前期男性相比，老年男性组L2－4、股骨颈、大转子的骨密度、E2、T、IL－1Ra/IL-1比值、BGP较低，而CTX较高，差异有显性，L2－4、股骨颈、Ward's骨密度、BGP与IL－1Ra/IL-1β呈正相关。CTX与IL－1Ra/IL-1β负相关、与IL－1β正相关。L2－4、大转子的骨密度、BGP与E2正相关；股骨颈、大转子骨密度、BGP与T正相关。IL－1Ra、IL－1Ra/IL-1β与E2正相关（P＜0．05）。结论 老年男性骨代谢处于高分解、低合成的负平衡状态，骨密度低于老年前期的男性，性激素降低影响IL－1Ra、IL－1两的平衡的可能起重要作用。     

白细胞介素1受体拮抗剂对人肾成纤维细胞增殖及产生纤连蛋白的影响

目的：研究白细胞介素1受体拮抗剂（IL－1β诱导人肾成纤维细胞（KFB）增殖及产生纤连蛋白（FN）的影响。方法：体外培养KFB,经IL－1β刺激后,分别采用^3H-TdR掺入法和ELISA法测定KFB的增殖及其FN的分泌。结果IL－1β（25－100ng/ml)可促进KFB增殖（P＜05）及分泌FN（P＜0．05）,IL－Ira(125-1000ng/ml)及IL－1ra(250-1000ng/ml)可以分别抑制IL－1β（50ng/ml)引起的KFB增殖（P＜0．05）及FN分泌（P＜0．05）。结论：IL-1β在肾间质纤维化中起了一定的致病作用,IL-lra sk c beggon IL-β的作用,从而为IL－l防治肾间质纤维化提供一定的实验依据。     

Successful treatment of Erdheim-Chester disease by interleukin-1 receptor antagonist protein

Erdheim-Chester disease is a rare non-langerhans systemic histiocytosis of unknown origin, associated with bone diseases and severe visceral complications. Therapies have been disappointing. A recombinant form of interleukin-1 receptor antagonist (anakinra) has been used in a few cases when usual treatment fails. We report a new case of successfully interleukin-1 receptor antagonist treatment in Erdheim-Chester disease.     

Serum levels of interleukin-1 receptor antagonist (IL-1ra) in thyroid cancer patients

Background and aims There is growing evidence that cytokines and their antagonists are important in the pathogenesis of various malignancies. While there are several reports on interleukin-1 receptor antagonist (IL-1ra) gene polymorphism and tissue expression, there is only little data available on the impact of IL-1ra serum levels. Therefore, we performed a prospective study, analyzing IL-1ra in thyroid cancer patients. Materials and methods We measured preoperative IL-1ra serum levels of 52 consecutive patients with thyroid cancer, 15 with benign adenoma and 27 healthy volunteers. The final histological diagnosis revealed 21 patients with papillary and 8 patients with follicular carcinoma (FTC), while 12 cases of medullary and 11 cases of anaplastic carcinoma (ATC) were observed. Results Compared to the control group, serum concentrations of IL-1ra were significantly higher in ATC and FTC patients. Concerning gender differences, this effect reached significance only in women with ATC and FTC. Except for the stage IV disease in ATC, there was no correlation between IL-1ra levels and International Union Against Cancer staging. Conclusion The findings of our study indicate that IL-1ra may play an important role in the development of ATC and FTC. Future efforts should focus on the possible application of IL-1ra as a biomarker for the above-mentioned thyroid malignancies.     

Persistent bone-sparing effect of interleukin-1 receptor antagonist: A hypothesis on the role of IL-1 in ovariectomy-induced bone loss

The recent finding that treatment with the interleukin-1 (IL-1) inhibitor, interleukin-1 receptor antagonist (IL-1ra) decreases bone loss and bone resorption in ovariectomized rats, strongly suggested that IL-1 mediates, at least in part, the effects of estrogen deficiency on bone resorption. Although in vitro studies have shown that IL-1 activates mature osteoclasts and stimulates osteoclastogenesis, the two main mechanisms by which estrogen deficiency stimulates bone resorption, it is still unclear whether IL-1 mediates both effects of estrogen deficiency in vivo. To investigate this matter, we have examined the changes in bone mineral density (BMD) which occur in ovariectomized rats after completion of 1 month of estrogen or IL-1ra treatment begun at the time of ovariectomy. Ovariectomy caused a marked decreased in BMD which was blocked by 17 estradiol and decreased by IL-1ra. Cessation of estrogen therapy was followed by a rapid induction of bone loss, indicating that estrogen blocks the activation and utilization of mature osteoclasts without depleting the bone microenvironment of osteoclast precursors and mature, inactive osteoclasts. In contrast, ovariectomized rats treated with IL-1ra maintained a stable bone density for the first 4 weeks after completion of the treatment. In these rats, bone loss resumed not earlier than 6 weeks after discontinuation of the IL-1ra treatment. Estrogen deficiency was necessary to unveil the bone-sparing effect of IL-1ra because in a control experiment in which rats were treated with IL-1ra for the 4 weeks before ovariectomy, BMD began to decrease immediately after ovariectomy. Based on these results we propose the hypothesis that in conditions of estrogen deficiency, the main effect of IL-1ra is to block the proliferation and differentiation of osteoclast precursors, an event that results in the depletion of mature, rapidly responsive osteoclasts. We also suggest that estrogen may have important direct effects on the regulation of osteoclast activity.     

Circulating interleukin-1 receptor antagonist (IL-1RA) serum levels in patients undergoing orthotopic heart transplantation

In a pilot study we determined the serum levels of circulating interleukin-1 receptor antagonist (IL-1ra) in patients undergoing orthotopic heart transplantation and in control patients scheduled for open heart surgery without allograft transplantation. Blood samples were obtained from 12 transplant recipients and 7 controls prior to the operative procedures to determine baseline values. Serum levels of IL-1ra were measured within 12 h of decrossclamping of the aorta and every 24 h for the following 14 days. Endomyocardial biopsies were obtained weekly for the 1st month after transplantation. Compared to baseline values, IL-1ra serum levels 12 h after decrossclamping of the aorta were significantly higher both in the control group (507 ± 165 vs 3980 ± 452 pg/ml, P < 0.01) and among the transplant recipients (413 ± 180 vs 4117 ± 459 pg/ml, P < 0.01) IL-1ra levels remained significantly elevated for 2 and 5 days, respectively. There were no significant differences in the IL-1ra serum levels between the two groups throughout the observation period. Endomyocardial biopsies of two patients showed acute allograft rejection, Billingham grade III a and III b, respectively. In both cases, the rejection episodes were accompanied by a renewed and more pronounced elevation in the IL-1ra serum levels beyond 4000 pg/ml for at least 2 days. These preliminary results indicate that IL-1ra may be a nonspecific immune marker during the first few days after orthotopic heart transplantation and cardiopulmonary bypass. Moreover, renewed, prolonged increases in IL-1ra appear to be associated with rejection. Further studies are needed to confirm the predictive value of IL-1ra in the detection of acute allograft rejection. Received: 26 May 1998 Received after revision: 21 August 1998 Accepted: 21 August 1998     

Induction of interleukin-1 and interleukin-1 receptor antagonist during contaminated in-vitro dialysis with whole blood

BACKGROUND: Previous studies on the permeability of cellulosic and syntheticdialysers for bacterial-derived cytokine-inducing substancesgave conflicting results. We tried to study this issue as closeto the in-vivo situation as possible. METHODS: An in-vitro dialysis circuit with whole human blood presentin the blood compartment of cuprophane (Cup), polysulphone (PS),and polyamide (PA) dialysers was employed; sterile filtratesderived from Pseudomonas aeruginosa cultures were added to thedialysate. We studied the induction of interleukin-1ß(IL-1ß) by plasma samples taken from the blood compartmentas well as the induction of IL-1ß and interleukin-1receptor antagonist (IL-1Ra) in mononuclear cells separatedfrom whole blood after circulation by radioimmunoassay and polymerasechain reaction. RESULTS: Plasma samples from the blood side of all dialysers inducedIL-1ß from non-circulated mononuclear cells afteraddition of pseudomonas filtrates to the dialysate; the maximalamount of IL-1ß induced by samples from the bloodcompartment was 4.8±1.2 ng/ml for Cup, 1.9±0.5ng/ml for PS, and 2.0±0.6 ng/ml for PA. Mononuclear cellsseparated after contaminated dialysis with all types of dialysersexpressed increased mRNA levels for IL-1ß and IL-1Ra.Production of IL-1Ra by cells separated after contaminated dialysiswas determined after Cup and PS dialysis; there was increasedproduction of IL-1Ra by these cells (Cup, 10.3±4.2; PS,7.3±2.5 ng/ml) compared to cells separated after steriledialysis (Cup, 5.6±2.1, P<0.05; PS, 4.5±1.1ng/ml, n.s.) or from non-circulated blood (Cup experiments,4.7±1.5, P<0.05; PS experiments, 4.1±1.2 ng/ml,n.s.). CONCLUSIONS: These data suggest penetration of cytokine-inducing substancesthrough both cellulosic and synthetic dialysers. Differencesbetween dialysers may exist regarding extent and time courseof penetration. The detection of cytokine mRNA as well as themeasurement of IL-1Ra synthesis is more sensitive substancesthrough dialyser membranes than the measurement of IL-1ßprotein synthesis.     

双后肢大鼠椎间盘退变动物模型的建立

目的　为椎间盘相关研究建立一种经济科学的椎间盘退变动物模型。方法　对 15 2只新生SD大鼠采用截除双前肢和特殊饲养的方法培育双后肢大鼠 ,以 10只正常同龄大鼠作为对照 ,18月龄时处死后通过光镜和电镜检查来观察两组大鼠L2～ 3 椎间盘的退变情况。结果　截除双前肢后 ,双后肢大鼠生长良好 ,术后 18个月存活 17只。光镜检查证实其椎间盘均发生了严重的髓核和纤维环退变。有 2例还出现了腰椎间盘突出。超微结构观察发现髓核中脊索细胞出现明显的退变和坏死 ,软骨样细胞中出现大量脂滴。基质中胶原纤维排列紊乱 ,出现大量板层样结构和团块。而正常大鼠的椎间盘仅发生轻度退变 ,胶原纤维排列整齐。结论　本造模方法简单经济、成功率高、重复性好 ,建立的双后肢大鼠模型符合人体椎间盘退变规律。     

白细胞介素-6对椎间盘细胞中蛋白多糖代谢影响的实验研究

目的:研究白细胞介素-6(interleukin-6,IL-6)对椎间盘纤维环和髓核细胞中蛋白多糖代谢的影响。方法:自然流产的胎儿4例,4h内无菌取出椎间盘,分别进行纤维环和髓核细胞的体外培养。在纤维环细胞的培养液中分别加入IL-60(对照组)、400、800ng/ml,培养24h后,用Alcian法检测培养液中硫酸软骨素的含量。在培养中的髓核细胞中加入IL-60(对照组)、100、400、800ng/ml,培养24h,然后测量培养液中硫酸软骨素的含量。结果:在纤维环细胞中加入IL-6组较不加IL-6对照组培养液中硫酸软骨素的含量增加,在髓核细胞组中加入IL-6组和对照组的差别无显著性。结论:IL-6可以刺激椎间盘纤维环细胞中蛋白多糖的合成,但对髓核细胞中蛋白多糖的合成没有明显的作用。     

Administration of interleukin-1 receptor antagonist ameliorates renal ischemia-reperfusion injury.

Interleukin (IL)-1 is a major contributor to inflammation and apoptosis during ischemia/reperfusion (I/R) injury. Its deleterious effects are primarily mediated by the activation of nuclear factor-kappaB (NF-kappaB). Receptor-binding and signaling of IL-1 can be blocked by the IL-1 receptor antagonist (IL-1ra). The aim of our study was to characterize effects and mechanisms of IL-1ra administration on inflammation, apoptosis, and infiltration in renal I/R injury. Renal ischemia was induced in Lewis rats by clamping of the left renal artery for 45 min. Kidneys were removed for histological and molecular analysis 24 h or 5 days after reperfusion. IL-1ra ameliorated I/R induced renal injury and inflammation. Furthermore, the number of apoptotic tubular cells was lower in IL-1ra-treated animals 24 h after ischemia, which was paralleled by a Bax/Bcl-2 mRNA ratio towards anti-apoptotic effects. IL-1ra reduced the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA at 24 h and 5 days and that of intracellular adhesion molecule-1 (ICAM-1) expression at 24 h in the ischemic reperfused kidneys. Our results indicate that IL-1ra treatment ameliorates renal I/R injury and this protective effect might be mediated by reduced induction of NF-kappaB mediated MCP-1, ICAM-1, and a decreased ratio between Bax and Bcl-2 mRNA expression.