霉酚酸酯对MRL-lpr/lpr小鼠狼疮性肾炎的治疗作用

目的　采用MRL lpr/lpr自发狼疮小鼠观察霉酚酸酯 (MMF)对狼疮性肾炎的治疗作用 ,探讨其作用机制。方法　 12周龄雌性MRL lpr/lpr自发狼疮小鼠分别予以MMF(90mg·kg-1·d-1,po)、甲基强的松龙 (MPS) (2 5mg·kg-1·d-1,ip)、环磷酰胺 (CTX) (2 5mg·kg-1·w-1,ip)治疗 2 4周。分别测定3种不同药物对小鼠尿蛋白排泄量、动物死亡率的影响 ;放免法测定血清抗dsDNA抗体的变化 ;运用RT PCR和免疫组化方法观察治疗药物对小鼠肾组织表达ICAM 1、PAI 1的影响。结果　治疗 2 4周后 ,各治疗组与对照组相比 ,体重均有不同程度增长 (P <0 .0 5 ) ,尿蛋白排泄量明显下降 (P <0 .0 1) [治疗9个月时 ,对照组为 (11.9± 2 .8)mg/d ,MMF组 (5 .1± 1.2 )mg/d ,MPS组 (3.4± 0 .8)mg/d ,CTX组 (2 .9±0 .3)mg/d],动物累积死亡率明显降低 ,血清中抗dsDNA抗体水平均有一定程度的降低 (P <0 .0 1) [治疗 9个月后 ,对照组为 (6 1.6± 7.1) % ,MMF组 (2 8.7± 4.6 ) % ,MPS组 (37.6± 4.3) % ,CTX组 (4 0 .1± 5 .8) % ],肾间质炎细胞浸润、肾小球硬化及间质纤维化程度较对照组减轻 ;肾组织免疫组化、RT PCR结果表明 ,3种药物在基因转录、蛋白质表达水平可不同程度地降低肾组织内ICAM 1、PAI 1表达。结论　MMF对MRL lpr/lpr自发     

近端胃功能与功能性消化不良症状的关系

探讨近端胃的顺应性及感觉阈值与功能性消化不良 (functionaldyspepsia ,FD)患者消化不良症状发生间的关系。利用电子恒压器 (SVS/Barostat)对 2 2例FD患者及 8例健康志愿者 ,分别测出近端胃的顺应性、初始容积和压力及最大耐受容积和压力。FD组及对照组的顺应性分别为 (4 4. 4± 15.0 )mL/mmHg(1mmHg =133.32 2Pa)、(6 6.7± 7.7)mL/mmHg,2组的最大耐受容积分别是 (6 4 6. 6± 14 5.1)mL、(76 8.8± 4 7.0 )mL ,FD组的顺应性及最大耐受容积明显低于对照组 (P <0.0 5 )。按消化不良症状积分分成轻、中、重度 3组 ,中、重度组的顺应性 [(4 0 .8± 17.6 )mL/mmHg、(34.9± 12.4 )mL/mmHg]和最大耐受容积 [(6 30. 0± 92.3)mL、(5 87.5± 89.7mL) ]均低于轻度组 [(5 0.4± 35.4 )mL/mmHg、(70 5.8± 195.3)mL](P <0.0 5 )。按早饱症状积分分为轻、中、重度 3组 ,轻、中度组的初始容积 [(36 8.3±114.1)mL、(35 5.8± 6 3.8)mL]和最大耐受容积 [(5 90.0± 2 9.3)mL、(5 4 7.2± 87.9)mL]明显高于重度组 [(2 34.4±4 0.3)mL、(4 86. 7± 5 8.4 )mL](P <0.0 5 )。早饱积分、腹胀积分分别与初始容积 (r=- 0.6 35 6 ,P <0.0 1;r=- 0.4 192 ,P <0.0 5 )以及消化不良症状积分与顺应性 (r=- 0.35 6 1,P <0.0 5 )之     

树鼩血栓性脑缺血时单胺氧化酶活性的变化及银杏内酯B作用机制探讨

目的 :揭示血栓性脑缺血时缺血区和血清单胺氧化酶 (MAO)活性变化及对血小板活化因子 (PAF)受体拮抗剂银杏内酯B(GB)作用机理的探讨。方法 :建立光化学诱导树鼠句血栓性脑缺血模型 ,用酶比色法测量缺血后4、2 4及 72h中心区、半暗区、对侧区及血清的MAO活性 ,并用双缩脲法测定上述各区的蛋白含量。结果 :脑缺血后不同时间缺血中心区MAO活性显著低于假手术组 [(15 4 1± 1 6 3)× 10 3 U/g蛋白 ]或对侧区 [(15 4 7± 1 6 8)× 10 3 U/g蛋白 ],以 72h最为明显 [(2 19± 1 96 )× 10 3 U/g蛋白 ,P <0 0 1];此时缺血半暗区和血清MAO活性 [分别为 (2 5 30± 2 0 1)× 10 3 U/g蛋白和 (2 10 0 4± 2 6 6 7)× 10 3 U/L]明显高于假手术组 (P <0 0 1)。光化学反应后 6h舌下静脉一次注射PAF受体拮抗剂GB(5mg·kg-1)后 2 4h时 ,半暗区MAO活性明显低于缺血组 ,而中心区则高于缺血组 (P <0 0 1)。MAO活性变化与相应区域蛋白含量改变一致 (r=0 81,P <0 0 5 )。结论 :脑缺血后中心区、半暗区MAO活性改变是相应区域单胺类递质消长变化的主要原因 ,MAO活性变化与神经元蛋白质合成能力的改变有关 ;GB的脑保护作用与其拮抗PAF受体和调节MAO活性而促进递质平衡有关。     

抗CD86单克隆抗体对哮喘小鼠TH1/TH2细胞因子的影响

目的观察抗CD86单克隆抗体对卵蛋白干粉(OVA)致敏并刺激小鼠TH1和TH2细胞因子比例的变化,为防治哮喘提供实验依据.方法经OVA致敏的雌性Balb/c于激发前腹腔注射抗CD86单克隆抗体及同型对照IgG2α 50 μg,末次激发48 h后收集支气管肺泡灌洗液(BALF),以酶联免疫吸附法(ELISA)测定BALF中白细胞介素-4(IL-4)、白细胞介素-5(IL-5)和干扰素-γ(INF-γ)水平.结果以OVA致敏并激发的哮喘对照组小鼠BALF中IL-4、IL-5水平升高[分别为(64.23±3.91)pg/mL、(379.84±73.02)pg/mL],与PBS对照组[分别为(22±2)pg/mL、(245.75±50.01)pg/mL]比较,差异有显著性(P＜0.05);OVA致敏并激发的哮喘对照组小鼠BALF中INF-γ水平降低[(98.73±16.41)pg/mL],与PBS对照组[(218.35±48.63)pg/mL]比较,差异有显著性(P＜0.05).抗CD86单克隆抗体组小鼠BALF中IL-4、IL-5水平降低[分别为(35.78±4.88)pg/mL、(222.98±58.68)pg/mL],而INF-γ水平升高[(206.92±47.8)pg/mL],与哮喘对照组及同型对照IgG2α组(98.73±16.41)pg/mL、(102.32±15.49)pg/mL比较差异有显著性(P＜0.05).结论抗CD86单克隆抗体通过阻断共刺激信号,纠正TH1/TH2失衡,从而达到抗气道炎症的作用.     

乙型肝炎病毒基因型不同的患者肝功能相关指标及免疫功能变化分析

目的 探讨基因型不同的乙肝患者肝功能、病毒载量和免疫功能的差异及临床意义.方法 145例乙肝患者划分两个年龄段:小于35岁和大于35岁,全部应用实时荧光PCR法进行乙型肝炎病毒(HBV)基因分型检测;同时采用实时荧光PCR法检测血清HBV-DNA载量;用全自动生化分析仪和流式细胞仪检测各组肝功能相关指标及淋巴细胞亚群含量.结果 145例HBV患者仅为两种基因型,B基因型62例(42.76％),C基因型83例(57.24％).B型患者谷丙转氨酶(ALT)、谷草转氨酶(AST)和HBV-DNA均稍高于C型,分别是[(263.6±36.13) U/L比(243.1±37.69) U/L]、[(128.1±15.84) U/L比(123.6±19.1)U/L]和[(6.131±0.2133)比(5.875±0.1725)],进行统计学分析后差异均无统计学意义(P＞0.05),但划分年龄段后,大于35岁的患者,B型ALT、AST和HBV-DNA均显著高于C型(P＜0.05),分别是[(227.6±34.52) U/L比(144.8±19.92) U/L]、[(124.5±19.4) U/L比(79.79±12)U/L]和[(6.166±0.2582)比(5.228±0.2644)].淋巴细胞亚群分析可见,所有年龄段或大于35岁患者均是B型的CD4+T细胞比例显著低于C型[(32.97±0.95)％比(35.81±0.85)％](P＜0.05),CD8+T细胞显著高于C型[(30.19 ±0.97)％比(27.44±0.92)％](P＜0.05),而CD3+T细胞、B细胞、NK细胞在B型和C型患者中则差异无统计学意义(P＞0.05).结论 145例HBV患者B、C型的分布未见显著差异;肝功能、病毒载量及T细胞亚群在年龄较大的B、C两组患者中存在显著差异.     

低氧诱导因子-1α和内皮素-1基因在大鼠低氧性肺动脉高压中的表达

目的　探讨低氧诱导因子 1(HIF 1α)和内皮素 1(ET 1)基因表达在低氧性肺动脉高压发病过程中的变化和作用。方法　复制低氧性肺动脉高压大鼠模型 ,测定平均肺动脉压 ,弹力纤维染色显示腺泡内肺动脉 ,用放射免疫法测ET含量 ,原位杂交方法进行检测HIF 1αmRNA。结果　HIF 1αmRNA在低氧各组腺泡内肺动脉有表达 ,低氧 14d组 (0 2 5 6 9± 0 0 46 8)和低氧 2 8d组 (0 2 2 5 8±0 0 45 3)染色强于低氧 5d组 (0 145 5± 0 0 2 72 )和正常组 (0 110 9± 0 0 2 2 4) ;ET 1mRNA在低氧各组腺泡内肺动脉有表达 ,低氧 14d组 (0 412 2± 0 0 783)和低氧 2 8d组 (0 36 84± 0 0 72 9)染色强于低氧5d组 (0 2 0 17± 0 0 34 9)和正常组 (0 185 5± 0 0 36 1) ,HIF 1α和ET 1基因表达在H14d组和H2 8d组明显增加 (P <0 0 5 )。肺动脉血中ET 1含量在H14d组 [(15 8 78± 2 5 14)pg/ml]和H2 8d组 [(142 93± 2 3 38)pg/ml]明显高于H5d组 [(79 6 8± 12 5 4)pg/ml]和正常组 [(6 5 37± 10 82 )pg/ml](P <0 0 5 ) ;H14d组 [(34 0± 5 8)mmHg]和H2 8d组 [(2 9 0± 4 7)mmHg]的mPAP也明显高于H5d组[(19 0± 3 5 )mmHg]和正常组 [(17 0± 2 8)mmHg](P <0 .0 5 ) ,并且与肺动脉血中ET 1含量呈正比(rs=0     

罗红霉素对大鼠内毒素性血症的治疗作用

目的探讨罗红霉素对大鼠急性肺损伤模型的治疗作用.方法动物随机分为⑴正常对照组(NC),经胃灌入生理盐水10mL/kg;⑵内毒素组(LPS),静脉注射lipopolysaccharide(LPS,2mg/kg)建立大鼠急性肺损伤(ALI)模型,5min后灌胃生理盐水10mL/kg;⑶罗红霉素治疗组(Rx),静脉注射LPS2mg/kg5min后,经胃灌入罗红霉素20mg/kg.测定各组平均动脉压、动脉血氧分压、氧饱和度、二氧化碳分压、肺系数、肺水含量、肺通透性指数,观察肺组织病理学变化.结果注射LPS后大鼠平均动脉压立即下降,0.5min降至最低后回升,1.5h升高达峰值,之后逐渐下降,4h后LPS组大鼠平均动脉压[(56.64±7.45)mmHg]与NC组比较[(97.94±7.73)mmHg]显著下降(P＜0.01);而Rx组[(103.40±8.56)mmHg]变化不明显(P＞0.05).Rx组动脉血氧分压[(85.36±7.67)mmHg]、二氧化碳分压[(42.75±4.24)mmHg]和pH(7.37±0.03)分别与LPS组[(52.43±8.25)mmHg,(67.38±7.21)mmHg,7.28±0.06]相比有明显改善(P＜0.05),Rx组与LPS组相比肺系数(0.58±0.03vs0.67±0.04,P＜0.05);肺水含量(78.76±2.13vs82.81±3.87,P＜0.05);肺通透性指数(6.25±0.74vs12.47±0.89,P＜0.05)有明显下降,病理学观察Rx组肺组织病变减轻.结论罗红霉素能改善大鼠内毒素血症时血液动力学变化和气体交换能力,减轻大鼠内毒素造成的肺损伤,该治疗作用是否与罗红霉素调节机体细胞炎性因子与抗炎性因子平衡有关,尚需进一步探讨.     

内皮素和一氧化氮/一氧化氮酶检测对老年糖尿病与血管病变患者临床价值的研究

探讨内皮素(ET)、一氧化氮/一氧化氮酶( NO/NOS)在糖尿病与血管病变患者中的变化。ET采用放射免疫分析法,NO/NOS采用酶法。结果是:(1)糖尿病组ET水平(68.80±37.35)pg/mL)较对照组(41.70±21.50pg/mL)显著升高(P<0.001);NOS水平(2.75±1.49U/mL)明显高于对照组(1.12±0.56U/mL),P<0.01;NO水平(49.32± 16.32μmol/L)明显低于对照组(54.60 ±15.60μmol/L),P<0.05。(2)糖尿病合并血管病变组ET水(80.1140.25pg/mL)显著高于无并发症组(62.73±24.29pg/mL),P<0.001;NOS水平(4.02 ± 0.59U/mL)明显高于无并发症组(2.51±1.19U/mL),P<0.001;NO水平(42.25 ± 10.10/μmol/L)明显低于无并发症组(52.16±14.59mol/L,P<0.05;NO/NOS(11.99±7.05)明显低于无并发症组(26.9±13.15),P<0.001提示 ET、NO/NOS与糖尿病的发生和发展有关,联合检测不仅可作为糖尿病及其血管并发症的重要依据,还将有助于糖尿病合并血管病变的预防和治疗。     

白细胞介素-37在类风湿性关节炎患者血清表达水平的研究

目的 通过检测类风湿性关节炎(RA)患者血清中细胞因子白细胞介素(IL)-37、肿瘤坏死因子-α(TNF-α)、IL-18、炎性指标红细胞沉降率以及C-反应蛋白(C-reactionprotein,CRP)水平,观察RA患者临床数据包括压痛关节数,肿胀关节数以及DAS28评分等,探讨RA患者血清IL-37水平升高的意义以其在RA发病机制中可能的作用.方法 80例RA患者、80例健康对照患者,通过酶联免疫吸附试验(ELISA)法检测其血清中细胞因子水平.结果 RA患者血清中IL-37[(40.33±11.25)pg/mL]、TNF-α[(110.41 ±35.37) pg/mL]、IL-18[(121.73±29.22) pg/mL]水平以及红细胞沉降率(ESR)[(42.31±15.02) mm./h]、CRP[(38.31±17.22) mg/L]水平明显高于对照组IL-37[(18.21±5.72) pg/mL]、TNF-α[(30.19±6.82) pg/mL]、IL-18[(55.47±7.29) pg/mL]水平,组间差异有统计学意义(P＜0.05).IL-37的表达与TNF-α、IL-18水平呈正相关(相关系数r=0.981,P=0.001).结论 IL-37在RA患者体内高表达并与其它几种炎性因子的表达具有相关性,IL-37可能作为炎性抑制因子参与了RA的发生、发展.     

Egb761对迟发性运动障碍模型大鼠的预防和治疗作用及可能机制

目的:分析银杏叶提取物Egb761对迟发性运动障碍(TD)模型大鼠行为学和血清脑源性营养因子(BDNF)和总抗氧化能力(TAC)水平的影响,探索TD可能的防治机制。方法:将32只雄性Sprague-Daw ley(SD)大鼠随机分为对照组[腹腔注射生理盐水(NS)2 m L/kg+第5周始灌胃5 m L/kg NS]、TD组[腹腔注射氟哌啶醇(Hal)2 mg/kg+第5周始灌胃5 m L/kg NS]、Egb761预防组(PEgb761组)(腹腔注射Hal 2 mg/kg同时灌胃5 m L/kg Egb761溶液)、Egb761治疗组(T-Egb761组)(腹腔注射Hal 2 mg/kg+第5周始灌胃5 m L/kg Egb761溶液)。每组8只,共观察10周,每周第7天评定大鼠口周不自主运动(VCM)的严重程度。第10周末采用酶联免疫吸附法(ELISA)检测血清BDNF浓度,采用分光光度法检测血清TAC水平。结果:与对照组相比,第3周末TD和T-Egb761组VCM评分显著增加(P0.05),第5周末达峰值;第6周后T-Egb761组VCM评分逐步下降;第10周末T-Egb761组VCM评分(4.1±2.0)显著低于TD组(27.9±5.8)(P0.001),与对照组(3.5±1.9)无差异(P0.05);P-Egb761组与对照组10个评估点VCM评分无差异;第10周末,TD组血清BDNF水平[(6.9±1.0)pg/m L]低于对照组[(8.6±2.5)pg/m L]、T-Egb761组[(8.9±1.5)pg/m L]和P-Egb761组[(9.6±1.4)pg/m L];TD组TAC水平[(11.9±3.2)U/m L]低于对照组[(18.2±5.5)U/m L]和T-Egb761组[(19.4±4.4)U/m L](P0.05),与P-Egb761组差异无统计学意义(P0.05)。结论:Egb761可预防和显著缓解TD模型大鼠VCM症状,神经元的保护因素和自由基代谢异常可能在TD发生、发展过程中作用关键。     

Serum soluble interleukin-6 receptor in MRL/lpr mice is elevated with age and mediates the interleukin-6 signal

The characteristics of soluble interleukin-6 receptor (sIL-6R) in murine sera were examined. To investigate a relationship between serum sIL-6R level and autoimmune diseases, quantitative analysis of serum sIL-6R in MRL/lpr mice was performed by an enzyme-linked immunosorbent assay. The serum sIL-6R level in MRL/lpr mice of both sexes was below the detection limit (< 1.0 ng/ml) at 8 weeks of age, but it increased in accordance with age and reached 42 ± 9.3 ng/ml in female and 31 ± 13 ng/ml in male mice at 30 weeks of age. In MRL/+ mice, although an age-associated increase in serum sIL-6R level was observed, it was much less extensive than that in MRL/lpr mice. Elevated serum sIL-6R level at the age of 30 weeks was observed in female and male (NZB × NZW)F1 mice (32 ± 10 ng/ml and 17 ± 5.0 ng/ml, respectively), and male BXSB/Mpj Yaa mice (42 ± 18 ng/ml), suggesting that elevated serum sIL-6R in aged mice is one of the characteristics of autoimmune-prone mice. Quantitative analysis of serum IL-6 in MRL/lpr revealed that the serum sIL-6R level correlated well with the serum IL-6 level. We also showed that sIL-6R in the sera from MRL/lpr mice could mediate the IL-6 functions through the IL-6 signal-transducing receptor component gpl30, suggesting that elevated production of sIL-6R may partly contribute to development of autoimmune disease in MRL/lpr mice.     

Genetic dissection of the complex pathological manifestations of collagen disease in MRL/lpr mice

An MRL strain of mice bearing a Fas-deletion mutant gene, lpr, MRL/MpJ-lpr/lpr (MRL/lpr) develops collagen disease involving vasculitis, glomerulonephritis, arthritis and sialoadenitis, each of which has been studied as a model for polyarteritis, lupus nephritis, rheumatoid arthritis and Sjögren’s syndrome, respectively. Development of such lesions seems dependent on host genetic background since the congenic C3H/HeJ-lpr/lpr (C3H/lpr) mice rarely develop them. To identify the gene loci affecting each lesion, a genetic dissection of these complex pathological manifestations was carried out. First, histopathological features in MRL/lpr, C3H/lpr, (MRL/lpr × C3H/lpr) F1 intercross, and MRL/lpr × (MRL/lpr × C3H/lpr) F1 backcross mice were analyzed. Genomic DNA of the backcross mice were subjected to association studies by Chi-squared analysis for determining which polymorphic microsatellite locus occurs at higher frequency among affected compared to unaffected individuals for each lesion. As a result, gene loci recessively associated with each lesion were mapped on different chromosomal positions. We concluded that each of these lesions in MRL/lpr mice is under the control of a different set of genes, suggesting that the complex pathological manifestations of collagen disease result from polygenic inheritance.     

An immunomodulating anti-rheumatic drug, lobenzarit disodium (CCA): inhibition of polyclonal B-cell activation and prevention of autoimmune disease in MRL/Mp-lpr/lpr mice

In this study, we examined the effect of an immunoregulatory antirheumatic agent, lobenzarit disodium (CCA), on spontaneously developing glomerulonephritis in MRL/Mp-lpr/lpr (MRL/l) mice. Starting from 6 weeks of age, mice were given CCA orally 5 days a week at a dose of 2 or 10 mg/kg. A control group was given the same volume of distilled water. The CCA treatment suppressed the excretion of protein in the urine. At 40 weeks of age, the incidence of proteinuria was 10/10 in the controls, 6/10 in the 2-mg/kg treatment group, and 5/10 in the 10 mg/kg group. The life span was prolonged dose dependently. The 50% survival time was 33 weeks for the controls, 35.5 weeks for the 2-mg/kg group, and 41 weeks for the 10-mg/kg group. The serum levels of anti-ssDNA antibody, anti-TNP antibody, and rheumatoid factor (RF) of the Ig G isotype and immune complex were reduced compared with control group. But the antibodies of Ig M isotype were not reduced. The serum Ig G1, Ig G2, and Ig G3 were significantly lower in the CCA-treated mice than in the controls. But again the serum level of Ig M was unchanged. These effects of CCA may be based on the suppression of lymphadenopathy. CCA may correct abnormal B-cell growth and differentiation factor release by the MRL/l abnormal T cells. These results show that CCA inhibits the development of lupus nephritis in MRL/l mice through the amelioration of the abnormal immune response, polyclonal B-cell activation.     

三氧化二砷对MRL/lpr狼疮鼠T-bet/GATA3信号通路的影响

目的:本文旨在探讨三氧化二砷(arsenic trioxide,ATO)对MRL/lpr狼疮鼠T-bet/GATA3信号通路的影响及其作用机制。方法:将20周龄MRL/lpr狼疮鼠和正常C57BL/6J小鼠随机分组,分别接受ATO(0.4mg·kg~(-1)·d~(-1))和同体积生理盐水(NS)治疗2个月,分离脾脏淋巴细胞,然后用实时荧光定量PCR法检测T-bet、GATA3、IFN-γ、IL-4的mRNA水平及T-bet/GATA3 mRNA的比值,Western blot法检测T-bet和GATA3的蛋白表达,ELISA法检测血清中IFN-γ和IL-4的浓度。结果:(1)MRL/lpr狼疮鼠NS组T-bet、IFN-γ的mRNA及蛋白表达、Tbet/GATA3 mRNA比值均高于C57BL/6J小鼠NS组(P0.05),而GATA3、IL-4的mRNA及其蛋白表达均低于C57BL/6J小鼠NS组(P0.05);(2)在MRL/lpr狼疮鼠中,与NS组相比,ATO组T-bet、IFN-γ的mRNA及蛋白表达、T-bet/GATA3 mRNA比值均下降(P0.05),而2组中GATA3、IL-4的mRNA及蛋白表达无明显差异;(3)在C57BL/6J小鼠中,NS组和ATO组之间以上指标均无显著性差异。结论:ATO可能通过影响MRL/lpr狼疮鼠Tbet/GATA3信号通路,降低T-bet表达,从而减少Th1型细胞因子IFN-γ的表达和分泌。     

The effect of danazol in the MRL/lpr mouse model of autoimmune disease

The purpose of the study was to determine if danazol was efficacious in the treatment of lupus MRL/MpJ (lpr) mice, as measured by longevity, proteinuria and serum amyloid protein (SAP) levels. Danazol, administered at an oral dose of 100 mg/kg, significantly prolonged survival of female MRL/MpJ (lpr) mice but had no effect on the mortality of their male counterparts. Medication with danazol began 40 days after birth of the mice and resulted in a significant decrease in proteinuria in female but not male lupus mice. The concentration of SAP, an acute phase reactant, was significantly decreased in danzoltreated female lupus mice at 80, 100, 120, 140 and 160 days of age when compared to vehicle-treated control mice. SAP levels in male lupus mice treated with danazol were significantly lower than normal control levels only at the 120 and 160 day time points. Measurements of mortality, proteinuria and SAP concentration indicate that danazol at 100 mg/kg is orally active in the treatment of MRL/MpJ (lpr) female, but not male mice.     

Experimental treatment of autoimmune MRL-lpr/lpr mice with immunosuppressive compound FK506.

A newly developed immunosuppressive drug, FK506 (Fujisawa, Japan) is known to inhibit T-cell immunity. We have evaluated the action of this compound in MRL/lpr mice which develop a severe autoimmune disease. Eight-week-old female MRL/lpr of mice were treated subcutaneously with 2 mg/kg (high dose), 0.8 mg/kg (medium dose), 0.2 mg/kg (low dose) or solvent only (control) six times per week. Survival times of the mice were prolonged in the medium and the high dose treatment groups. The lymph node swelling was dramatically prevented with the high dose treatment. The increasing footpad swelling seemed to be also suppressed with the treatment. FACS analyses of the spleen cells revealed that FK506 reduced the percentage of double negative T cells (Thy-1.2+, Lyt-2-, L3T4-). Serological studies showed that anti-ssDNA and anti-dsDNA activities were significantly reduced by the high dose treatment, which is different from recent findings with Cyclosporine A. The high dose treatment also suppressed the total amount of IgG, even though the IgG concentration was rather increased by the medium dose treatment. Decreased proteinuria as well as pathological evaluations of the kidneys and lungs indicated that there were marked ameliorations in these organs with the treatment. These results suggest that FK506 could be potentially used for the treatment of autoimmune diseases.     

IL-33 Neutralization Suppresses Lupus Disease in Lupus-Prone Mice

IL-33 is a new member of the IL-1 family that plays a role in inflammation. In this study, we evaluated the potential of IL-33 inhibition as a treatment for systemic lupus erythematosus (SLE) using the lupus-prone model MRL/lpr mice and the underlying mechanisms of action. We treated mice with anti-mouse IL-33 antibody (anti-IL-33Ab) via intraperitoneal injection every other day from week 14 until week 20 for 6 weeks. A control group received the same amount of IgG control. Renal damage and mouse survival were compared. Cytokines, antibodies, immune complex, Tregs, myeloid-derived suppressor cells (MDSCs), and Th17 cells were also analyzed. Correlations between serum IL-33 and SLE disease activity index in human SLE were also investigated. MRL/lpr mice treated with anti-IL-33Ab showed reduced proteinuria and reduced serum anti-dsDNA levels. Nephritis, immune complex deposits, and the circulating antibodies and immune complex besides the mortality were significantly reduced by anti-IL-33Ab. Anti-IL-33Ab remarkably increased Tregs and MDSCs and reduced the Th17 cells and IL-1β, IL-6, and IL-17 levels in MRL/lpr mice. These results suggest that IL-33 inhibition may inhibit SLE via expansion of Tregs and MDSCs and inhibition of Th17 cells and proinflammatory responses, indicating that blockade of IL-33 has a protective effect on SLE.     

Short-Term Administration of Selected Anti-T-Cell Receptor Vβ Chain Specific MoAb Reduces Sialadenitis in MRL/lpr Mice

Sialadenitis develops spontaneously in MRL/Mp mice bearing a lymphoproliferative gene, lpr (MRL/ Mp- lpr/lpr ). Based on recent observations of an oligoclonal expansion of T-cell receptor (TCR) expressing Vβ chain families (Vβ4, Vβ8.1, 2, Vβ10b) in salivary glands of these mice we have initiated selective antibody therapy. Treatment with monoclonal antibodies (MoAb) specific for T cells expressing a mixture of TCR Vβ4, Vβ8.1, 2 and Vβ10b was applied to MRL/ lpr mice before and after the spontaneous development of Sialadenitis. The in vivo treatment with Vβ4, Vβ8.1, 2 and Vβ10b MoAb did not prevent the development of Sialadenitis. However, in animals with established Sialadenitis, treatment with the MoAb significantly decreased the inflammation compared with the control groups, Immuno-histochemical staining of cell phenotypes demonstrated a change in the ratio of CD4/CD8 in the animals with established Sialadenitis. Altogether, these findings illustrate that it is possible to modulate Sialadenitis and infiltrate cell phenotypes in vivo in MRL/ lpr mice with specific anti-TCR Vβ MoAb treatment.     

Induction of different types of glomerulonephritis by monoclonal antibodies derived from an MRL/lpr lupus mouse.

MRL/Mp-lpr/lpr(MRL/lpr) lupus mice develop glomerulonephritis in which the histopathological manifestations of the disease are characterized by diffuse cell-proliferative, crescentic, and/or wire loop-like lesions, resembling those of human lupus nephritis. Although these lesions are thought to be mediated by antibodies, little data is available to explain these regular variations in glomerular lesions induced by antibodies at the monoclonal level. We studied glomerular lesions of normal or severe combined immunodeficient mice injected with nephritogenic immunoglobulin G3-producing hybridoma clones (2B11.3 and 7B6.8), which we previously established from an unmanipulated MRL/lpr mouse. Both clones caused increased serum levels of immunoglobulin G3 with identical patterns over time and both induced glomerular deposits of immunoglobulin G3 and C3. However, 2B11.3 and 7B6.8 induced glomerular lesions that differed in their histopathological manifestations. The 2B11.3 clone generated cell-proliferative lesions associated with marked Mac-2-positive macrophage infiltrates, but the 7B6.8 clone induced lesions characterized by subendothelial hyaline deposits resembling wire loops. The latter was not associated with significant inflammatory cell infiltrates at any point throughout the progression of the lesion. Thus, our findings suggest that the histopathological variation in glomerulonephritis seen in MRL/lpr mice results from clonally expanded B cell clones that produce nephritogenic antibodies with different pathogenic potencies.     

三氧化二砷对MRL/lpr狼疮鼠IFN-γ基因表达的调控

目的: 探讨三氧化二砷(ATO)调控MRL/lpr狼疮鼠干扰素γ(IFN-γ)基因表达的机制。方法: 将20周龄MRL/lpr狼疮鼠和正常C57BL/6J小鼠无菌条件下取出脾脏,制成脾脏淋巴细胞悬液。体外经植物血球凝集素P(PHA-P,终浓度20 mg/L)和白细胞介素-2(IL-2,终浓度10⁶ IU/L)常规刺激48 h后,随机分为PBS组(空白对照)和 ATO(1.0 μmol/L)组,继续培养24 h。酶联免疫吸附法(ELISA)测定各组培养上清液IFN-γ的表达量,采用实时荧光定量PCR(Q-PCR)检测各组IFN-γ mRNA的表达情况,应用基于半定量PCR和Q-PCR的染色质免疫共沉淀(ChIP)技术检测各组细胞 IFN-γ 启动子区乙酰化组蛋白H3、H4(acH3, acH4)及启动子区结合RNA聚合酶Ⅱ(RNAPⅡ)的水平。结果: (1)MRL/lpr狼疮鼠PBS组IFN-γ的分泌和IFN-γ mRNA的表达均高于C57BL/6J小鼠PBS组(分别P<0.01和 P<0.05),并且 IFN-γ 启动子区acH3、acH4的水平及启动子区域富集RNAPⅡ水平高于C57BL/6J小鼠PBS组(均P<0.01)。(2)在MRL/lpr狼疮鼠中,与PBS组相比,ATO组IFN-γ的分泌和IFN-γ mRNA的表达下降(分别P<0.01, P<0.05),IFN-γ基因启动子区域acH3、acH4的水平及启动子区域富集RNAPⅡ水平也下降(均P<0.01)。(3)在C57BL/6J小鼠中,PBS组和ATO组之间以上指标均无差异。结论: ATO下调MRL/lpr狼疮鼠IFN-γ的分泌和IFN-γ mRNA的表达可能是通过降低基因启动子区域acH3、acH4的水平减弱了RNAPⅡ依赖的转录,而ATO对正常C57BL/6J小鼠无明显影响。