白介素—1受体拮抗剂对实验性新月 体肾炎骨调素表达的调节作用

探讨白介素－１受体拮抗对实验性新月体肾炎脏骨调不表达的调节作用，进一步阐明ＩＬ－１在实验性新月性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鼠基底膜肾毒血清制备。     

CD44及其配体骨调素在大鼠实验性新月体肾炎中的表达及作用

目的探讨ＣＤ４４及其配体骨调素（ＯＰＮ）在实验性新月体肾炎发展过程中的表达和作用。方法应用兔抗鼠肾毒性血清制作新月体肾炎模型，免疫组化双标记及原位杂交技术检测ＣＤ４４、ＯＰＮ及其它指标。结果ＣＤ４４和ＯＰＮ在新月体肾炎的发展过程中表达明显升高，并和巨噬细胞在肾脏组织的局部浸润、新月体的形成及肾功能的损害密切相关。结论在实验性新月体肾炎模型，ＣＤ４４和ＯＰＮ在肾小球实质细胞和小管间质细胞显著表达，并可能在新月体肾炎的发病过程中发挥重要的作用。     

骨调素对巨噬细胞在新月体肾炎肾脏组织局部浸润的影响

目的观察骨调素（ＯＰＮ）在大鼠实验性新月体肾炎肾脏组织中的表达及功能性抑制ＯＰＮ的活性对巨噬细胞在肾脏局部浸润的影响。方法ＯＰＮ在肾脏组织蛋白和基因水平的表达分别应用免疫组化双标记及原位杂交技术。结果在实验性新月体肾炎肾脏组织内ＯＰＮ的表达明显升高，并与肾脏局部巨噬细胞的浸润、肾脏病理损害程度及肾功能的下降明显相关。应用抗ＯＰＮ抗体治疗以后可显著抑制ＯＰＮ在肾脏组织的表达，同时巨噬细胞在肾脏组织局部的浸润也明显减少，肾脏病理损害减轻、肾功能明显改善。结论ＯＰＮ的高表达是实验性新月体肾炎发病过程中巨噬细胞在肾脏组织局部浸润的重要介导因子，抑制ＯＰＮ的表达和功能可显著抑制肾脏局部巨噬细胞的聚集及改善肾功能。     

白介素-1受体拮抗剂对椎间盘基质代谢的影响

目的:探讨白介素-1受体拮抗剂(IL-1Ra)对双后肢大鼠椎间盘基质代谢的影响。方法:建立双后肢大鼠模型36只,随机分为实验组和对照组,每组18只,实验组从造模当天即予腹腔注射IL-1R(a50ng/kg),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只大鼠,完整取出L4/5椎间盘固定、切片,予SABC法进行免疫组化染色,并使用图像分析系统对纤维环中Ⅱ型胶原染色进行灰度值扫描,同理取出L5/6椎间盘,使用间苯三酚法测量蛋白多糖含量。结果:术后第1个月,实验组椎间盘髓核中Ⅱ型胶原灰度值及蛋白多糖含量分别为83.67±7.97和2.376±0.161,对照组分别为87.25±8.70和2.297±0.101,两组间无显著性差异(P>0.05);第3个月时实验组Ⅱ型胶原染色的灰度值及蛋白多糖含量分别为107.42±6.50和2.093±0.131,对照组分别为145.91±7.42和1.039±0.092,两组间比较有非常显著性差异(P<0.01);第6个月时实验组Ⅱ型胶原染色的灰度值及蛋白多糖含量分别为129.83±4.03和1.796±0.065,对照组分别为203.76±5.98和0.654±0.048,两组间比较有非常显著性差异(P<0.01)。结论:IL-1Ra对双后肢大鼠椎间盘基质降解有明显的抑制作用。     

反义寡核苷酸对间质性肾炎肾小管上皮细胞骨调素表达的调节

目的 探讨骨调素（ＯＰＮ）反义寡核苷酸对间质性肾炎肾小管上皮细胞ＯＰＮ表达的影响。方法 以一侧输尿管梗阻（ＵＵＯ）所致的间质性肾炎为研究模型,肾动脉内注射全硫代修饰的ＯＰＮ反义寡核苷酸,免疫组织化学双染及原位杂交技术检测ＯＰＮ的表达及巨噬细胞的浸润。结果 ＯＰＮ及其ｍＲＮＡ在间质性肾炎的发展过程中表达明显升高,巨噬细胞的局部浸泣均发生在过度表达ＯＰＮ的肾小管周围;ＯＰＮ反义寡核苷酸能显著抑制肾小管     

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。     

黑皮素-1受体对皮肤色素的调控作用

获得性皮肤色素沉着或色素脱失是整形美容外科常见的现象，目前还没有有效的方法解决此类问题，主要原因之一是对于调控皮肤色素沉着的机制还不完全清楚，随着黑皮素受体克隆的成功，人们对皮肤色素沉着的机制有了进一步的了解，下面我们对黑皮素-1受体（melanoeontin-1 receptor,MC-IR)在皮肤色素沉着中的调控作用作一综述。     

不同类型新月体肾炎患者肾小球中C-C类趋化因子的表达及其意义

目的：研究不同类型新月体肾炎患者肾小球中C－C类趋化因子单核细胞趋化蛋白－1（MCP－1）、巨噬细胞炎性蛋白－1α和β（MIP－1α、MIP－1β）的表达,分析其与肾小球中浸润的单核细胞及Bowman囊壁是否断裂之间的关系,从而探讨其在新月体肾炎发生和发展中的作用。方法：选择不同类型新月体肾炎患者32例,其中I型8例,Ⅱ型12例,Ⅲ型12例。应用免疫组织化学方法分别观察细胞性和纤维细胞性新月体肾炎的肾小球中CD68、MCP－1、MIP－1α和MIP－1β的表达。结果：3种类型新月体肾炎肾小球中CD68、MCP－1、MIP－1α和MIP－1β的表达均明显增加;新月体肾炎肾小球中MCP－1和MIP－1α的表达与单核细胞浸润之间呈正相关（相关系数r分别为0．568和0．749,P均＜0．01）;I型新月体肾炎不仅肾小球中MCP－1和MIP－1α的表达均高于Ⅱ型和Ⅲ型新月体肾炎。而且Bowman囊壁断裂的发生率也较Ⅱ型和Ⅲ型新月体肾炎患者明显升高（P＜0．05）。结论：肾小球内MCP－1、MIP－1α和MIP－1β表达增加是新月体形成的重要始动因素,MCP－1和MIP－1α不仅介导新月体肾炎肾小球中单核细胞的浸润,而且可能还与Bowman囊壁断裂的发生有关。不同类型新月体肾炎患者肾小球内上述趋化因子表达的差异及由此所致病变程度的不同,可能直接影响到患者的预后。     

反义骨调素寡核苷酸对大鼠肾系膜细胞骨调素mRNA表达及细胞黏附能力的影响

目的 观察反义骨词素(OPN)寡核苷酸(AS—ODN)对大鼠肾系膜细胞(1097)骨调素mRNA表达及细胞黏附功能的影响。方法 针对OPN的cDNA序列设计合成一条与OPNcDNA序列互补的AS—ODN，所有碱基均经硫代修饰并将寡核苷酸与阳离子脂质体(DOTAP)结合。不同浓度寡核苷酸处理1097细胞48h，用Trizol试剂提取细胞总RNA。采用RNA斑点杂交和RT-PCR技术检测OPNmRNA的表达。采用胶原凝胶黏附法检测细胞的黏附功能。结果 在无血清条件下培养，1097细胞OPNmRNA表达水平很低，加入小牛血清后，其OPNmRNA表达水平明显上调。反义OPN寡核苷酸处理的1097细胞OPNmRNA表达水平较低，明显低于1640培养基对照组，且具有AS-ODN的浓度依赖性，其最低抑制浓度为2．5μmol／L。而经顺义OPN寡核苷酸或错义OPN寡核苷酸处理的1097细胞OPNmRNA表达水平仍然较高，无明显抑制作用。反义OPN寡核苷酸处理的细胞黏附胶原凝胶的能力较弱，易被洗涤下来；而顺义或错义OPN寡核苷酸处理的细胞黏附凝胶的能力仍然较强。结论 小牛血清能诱导大鼠肾系膜细胞OPN的上调。反义OPN寡核苷酸能特异性地抑制大鼠肾系膜细胞OPNmRNA的表达和对胶原凝胶的黏附能力。     

Stimulation of PAI-1 in rabbit anti-GBM glomerulonephritis

It has previously been shown in human disease and animal models of glomerulonephritis (GN) that fibrin deposition is associated with a net reduction of glomerular fibrinolytic activity as a result of reduced expression of plasminogen activators and increased expression of plasminogen activator inhibitor type 1 (PAI-1). Conditioned media (CM) prepared from cultured glomeruli of normal rabbits and rabbits 24 (Day 1) and 96 (Day 4) h after induction of anti-GBM GN were compared for their effects on the synthesis of fibrinolytic molecules in human endothelial cells (EC). Only CM from Day 4 GN rabbits showed PAI-1 protein stimulatory activity of up to 148% ( P <0.05; n =3) above that of untreated EC. This was also seen at the mRNA level. Glomerulonephritis Day 4 CM showed significantly higher amounts of tumour necrosis factor (TNF) and thrombin and transforming growth factor-β (TGF-β) bioactivity in comparison to glomerular CM from normal rabbits. After high performance liquid chromatography (HPLC) of Day 4 GN CM, PAI-1 stimulatory activity was found to correlate with the presence of interleukin 1 (IL-1), TNF and TGF-β. These results suggest a correlation between severity of anti-GBM GN in a rabbit model, increased PAI-1 synthesis and increased expression of TNF and TGF-β. This may potentiate glomerular fibrin and extracellular matrix deposition in anti-GBM GN, leading to glomerular crescent formation and eventual renal failure.     

Stimulation of PAI-1 in rabbit anti-GBM glomerulonephritis

SUMMARY: It has previously been shown in human disease and animal models of glomerulonephritis (GN) that fibrin deposition is associated with a net reduction of glomerular fibrinolytic activity as a result of reduced expression of plasminogen activators and increased expression of plasminogen activator inhibitor type 1 (PAI-1). Conditioned media (CM) prepared from cultured glomeruli of normal rabbits and rabbits 24 (Day 1) and 96 (Day 4) h after induction of anti-GBM GN were compared for their effects on the synthesis of fibrinolytic molecules in human endothelial cells (EC). Only CM from Day 4 GN rabbits showed PAI-1 protein stimulatory activity of up to 148% ( P <0.05; n = 3) above that of untreated EC. This was also seen at the mRNA level. Glomerulonephritis Day 4 CM showed significantly higher amounts of tumour necrosis factor (TNF) and thrombin and transforming growth factor-β (TGF-β) bioactivity in comparison to glomerular CM from normal rabbits. After high performance liquid chromatography (HPLC) of Day 4 GN CM, PAI-1 stimulatory activity was found to correlate with the presence of interleukin 1 (IL-1), TNF and TGF-β. These results suggest a correlation between severity of anti-GBM GN in a rabbit model, increased PAI-1 synthesis and increased expression of TNF and TGF-β. This may potentiate glomerular fibrin and extracellular matrix deposition in anti-GBM GN, leading to glomerular crescent formation and eventual renal failure.     

Important role for macrophages in induction of crescentic anti-GBM glomerulonephritis in WKY rats.

BACKGROUND: A crucial role for CD8(+) cells in induction of crescentic anti-glomerular basement membrane (GBM) glomerulonephritis (GN) in WKY rats was demonstrated in studies showing that depletion of CD8(+) cells completely suppressed glomerular accumulation of monocytes/macrophages (Mo/Mphi), crescent formation and proteinuria. Because these studies did not definitively identify CD8(+) cells as the cause of tissue injury, we examined the roles of Mo/Mphi in the development of anti-GBM GN. METHODS: We examined correlations between the amount of urinary protein and the numbers of glomerular CD8(+) cells or Mo/Mphi in rats after administrating different doses of anti-GBM antibody (5.0, 7.5, 10.0 and 25.0 microl/100 g body weight). The roles of Mo/Mphi in induction of GN were examined in animals by depleting Mo/Mphi in the glomerulus. To do this, rats were injected intravenously with liposome-encapsulated dichloromethylene diphosphonate (liposome-MDP) from day 3 to day 7 after anti-GBM antibody injection and they were then sacrificed at day 8. RESULTS: Liposome-MDP treatment significantly reduced the number of ED-1(+) Mo/Mphi accumulated in glomeruli from 32.1 +/- 1.2 to 1.4 +/- 0.3/glomerular cross-section (mean +/- SD, P < 0.01), and the amount of urinary protein from 103.8 +/- 19.8 to 31.8 +/- 15.9 mg/day (P < 0.01), as well as the incidence of crescentic glomeruli from 91.3 +/- 2.7 to 23.3 +/- 7.6% (P < 0.01) at day 8. This treatment also reduced the number of CD8(+) cells accumulating in the glomeruli from 5.4 +/- 0.7 to 0.5 +/- 0.1/glomerular cross-section (P < 0.01). Upregulation of glomerular intercellular adhesion molecule 1 (ICAM-1) and monocyte chemoattractant protein 1 (MCP-1) mRNA expression was suppressed by Mo/Mphi depletion. CONCLUSION: These results indicate that Mo/Mphi play an important role in the induction of crescentic anti-GBM GN and glomerular injury.     

白细胞介素1受体拮抗剂等位基因与紫癜性肾炎和IgA肾病的相关联系

目的 为了进一步阐明紫癜性肾炎(HSPN)及IgA肾病(IgAN)二者在发病机理上可能存在的联系。方法 用PCR方法对43例HSPN,97例IgAN患者和98名正常人IL-l受体拮抗剂(IL-lra)基因数目可变的串联重复(VNTR)多态性进行了分析。结果 HSPN患者白细胞介素l受体拮抗剂等位基因(IL IRN 2)的携带率明显高于正常人(P<0.02)和IgAN患者(P<0.05)。在IgAN患者中表现为反复发作性肉眼血尿者其IL IRN 2等位基因的携带率明显高于其它临床类型IgAN患者(P<0.o1),而与HSPN无显著性差异(P>0.05)。结论 HSPN患者IL-lra基因特定的遗传背景在其发病机理中可能起一定作用。IgAN患者中表现为反复发作性肉眼血尿者较其它临床类型IgAN患者与HSPN有更多的相似之处,而ILlRN 2等位基因高携带率可能是二者发病机理的共同点之一。     

Intrathecal co-administration of NMDA antagonist and NK-1 antagonist reduces MAC of isoflurane in rats

BACKGROUND: Intravenous administration of N-methyl-D-aspartate (NMDA) receptor antagonists and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists reportedly reduce the minimum alveolar anaesthetic concentration (MAC) for inhalation anaesthetics. If pain perception can be prevented by the intrathecal administration of antinociceptive receptor antagonists, these agents may reduce the requirements for inhalation anaesthetics. We studied the effect of intrathecal administration of an AMPA/kainate receptor antagonist, a metabotropic glutamate (mGlu) receptor antagonist and co-administration of NMDA and a neurokinin-1(NK-1) receptor antagonist drugs at low doses on the MAC. METHODS: After Wistar rats (n=36) were fitted with indwelling intrathecal catheters, the MAC of isoflurane was determined following intrathecal administration of a non-NMDA receptor antagonist (CNQX) at 10 microg, a mGlu receptor antagonist (AP3) at 10 microg, or a combination of NMDA receptor antagonist (APV) at 0.01 microg to 1 microg with NK-1 receptor antagonist (CP96345, CP) at 0.1 microg to 10 microg. Subsequently, a reversal dose of intrathecal NMDA with substance P (SP) was administered, and the MAC of isoflurane was redetermined. Conscious rats (n=15) were also examined for the presence of locomotor dysfunction following the intrathecal co-administration of APV and CP. RESULTS: Neither CNQX nor AP3 reduced the MAC of isoflurane. APV at 0.01 microg plus CP at 1 microg, as well as APV at 0.1 microg plus CP at 10 microg, reduced the MAC of isoflurane, with respective reductions of 7.6% and 14%; (P<0.05). Co-administration of NMDA plus SP reversed the decrease in the MAC of isoflurane. Locomotive activity was not changed. CONCLUSIONS: The NMDA receptor and the NK-1 receptor are important determinants of the MAC of isoflurane, exerting this influence by inhibition of pain transmission in the spinal cord, while mGlu and AMPA receptors have no effect on the MAC of isoflurane.     

The leukotriene B4 receptor antagonist ONO-4057 inhibits mesangioproliferative changes in anti-Thy-1 nephritis.

OBJECTIVE: ONO-4057 is a specific leukotriene B4 (LTB4) receptor antagonist which inhibits human neutrophil aggregation, chemotaxis and degranulation induced by LTB4. This study was conducted to evaluate the role of LTB4 in glomerulonephritis, and to examine whether ONO-4057 moderated anti-Thy-1 nephritis. METHODS: Experiment 1: Sixty Wistar rats were divided into three groups. Rats of Group A (n = 20) underwent intraperitoneal administration of placebo as a control group, rats of Group B (n = 20) first underwent intraperitoneal administration of 100 mg/kg ONO-4057 and rats of Group C (n = 20) first underwent intraperitoneal administration of 300 mg/kg ONO-4057 daily from day 3 before anti-Thy-1 antibody (OX7) injection to day 14 after OX7 injection, respectively. Experiment 2: Forty rats were divided into two groups. ONO-group (n = 20) was treated with 300 mg/kg BW of ONO-4057 and placebo-group (n = 20) with placebo daily from days 1 to 13 after OX7 injection. Urine and blood samples were collected and the kidneys were extirpated from five rats of each group sacrificed at 3 h, 24 h, day 7 or day 14 after the injection of OX7 in both experiments. Urinary protein excretion, renal function and pathological findings were analysed in each group of both experiments. RESULTS: (1) Glomerular infiltration by polymorphonuclear leucocytes (PMNs) and macrophages at 3 h was less in Groups B and C than in Group A, and matrix scores at day 7 were lower in Groups B and C than in Group A. Injury scores did not differ among the groups. (2) Urinary protein excretion at day 7 was less in Group C than in Group A. (3) Neither pathological findings nor urinary protein excretion differed between ONO-group and placebo-group. CONCLUSION: These results suggest that LTB4 is associated not with the pathogenesis of complement-dependent mesangial cell lysis but with that of mesangial proliferative change in anti-Thy-1 nephritis.     

Impact of clinical and histopathological factors on outcome of egyptian patients with crescentic glomerulonephritis

This study included 128 patients with crescentic glomerulonephritis (CGN) having sufficient clinical and histopathological data and were followed up in our institute for a mean period of 34 +/- 28 months. There were 49 males and 79 females with mean age 22.7 +/- 14 years. We studied the effect of clinical, laboratory and histopathological parameters on kidney function and patient survival at the end point of the study. The multivariate analysis revealed that serum creatinine at presentation, nephrotic range proteinuria during the follow up period, percentage of glomeruli affected by crescents, percentage of fibrous crescents and absence of cellular infiltration were significant risk factors affecting the kidney function at termination of the study. The only risk factor which correlated significantly with the patient mortality was the serum creatinine at last follows up.     

Glomerular expression of lipocortin-1 in human glomerulonephritis

Summary: Lipocortin-1 (LC-1), a Ca⁺⁺-dependent phospholipid binding protein, is believed to be involved in anti-inflammatory actions of glucocorticoids. to prove the hypothesis that steroid-resistant glomerulonephritis would show increased expression of LC-1, we evaluated the expression of LC-1 in various types of glomerulonephritis. Frozen samples of seven normal kidneys and 30 kidney biopsy tissues were stained with indirect immunofluorescent method. In the normal tissues, minimal change disease ( n =9), lupus nephritis ( n =5) and IgA nephropathy ( n =6), glomeruli did not stain for LC-1. Positive reactions for LC-1 were observed along the peripheral capillary walls in all five patients with membranous nephropathy with out hepatitis B surface antigen (HBsAg). In the patients with membranous nephropathy (MN) who also had chronic liver disease and HBsAg ( n =3), only weak reactions for LC-1 were found along the capillary walls and mesangial area in 1 patient. Patients with membranoproliferative glomerulonephritis ( n =2) showed positive reactions for LC-1 along the capillary walls. Fourteen patients with minimal change disease or lupus nephritis were treated with prednisolone. Ten patients showed substantial reduction of proteinuria, but four patients did not; however, staining for LC-1 was not negative in the kidney tissues of both steroid-responsive and steroid-resistant patients. These findings suggest that LC-1 does not mediate the action of glucocorticoids in human glomerulonephritis.