SLE病人PBMC分泌IL—6活性水平的研究

为了研究了ＳＬＥ病人ＰＢＭＣ分泌ＩＬ－６的能力，我们采用ＭＴＴ比色法，用ＩＬ－６依赖细胞株（７ＴＤ１）分别测定了１５例活动期，１５例非活动期ＳＬＥ病人和１５例正常对照者ＰＢＭＣ培养上清ＩＬ－６的活性水平。结果表明ＳＬＥ患者ＰＢＭＣ自发分泌ＩＬ－６的水平显著高于正常人（Ｐ〈０．０２或Ｐ〈０．００１），活动期患者高于非活动期患者（Ｐ〈０．００１）。ｐＨＡ－Ｍ刺激培养体系后，ＳＬＥ患者及正常人ＰＢＭＣ培     

硝酸甘油对哮喘患者一氧化氮内皮素的影响及机制

目的　了解哮喘患者肺泡巨噬细胞（ＡＭ） 、支气管上皮细胞（ＢＥＣ） 源性一氧化氮（ＮＯ）、内皮素（ＥＴ）的分泌状态及硝酸甘油（ＮＴＧ）对哮喘患者ＡＭ、ＢＥＣ产生ＮＯ、ＥＴ的影响及机制。方法　分离纯化了１５ 例轻、中度哮喘发作期患者、７ 名健康受试者ＡＭ、ＢＥＣ,并分为哮喘未干预组、哮喘ＮＴＧ干预组和健康对照组,用放射免疫法和镀铜镉还原法分别测定ＡＭ、ＢＥＣ培养４８ 小时上清液中ＥＴ、ＮＯ·２／ＮＯ·３ 浓度,用原位杂交的方法检测ＡＭ、ＢＥＣｉＮＯＳｍＲＮＡ、ＥＴｍＲＮＡ 的表达。结果　（１） 健康受试者ＡＭ、ＢＥＣ分泌少量ＮＯ和ＥＴ及少量ｉＮＯＳｍＲＮＡ 、ＥＴｍＲＮＡ表达;（２）哮喘患者ＡＭ、ＢＥＣ源性ＮＯ、ＥＴ水平及ＡＭ、ＢＥＣｉＮＯＳｍＲＮＡ、ＥＴｍＲＮＡ表达与各组比较差异有显著性（ Ｐ均＜ ０－０５）;（３）ＮＴＧ 促进哮喘患者ＡＭ、ＢＥＣ源性ＮＯ产生（ Ｐ均＜０－０５）,明显抑制ＥＴ产生和ＥＴｍＲＮＡ 的表达,与对照组比较差异均无显著性（ Ｐ均＞ ０－０５） ,ＮＴＧ同时抑制哮喘患者ＡＭ、ＢＥＣｉＮＯＳｍＲＮＡ的表达,与健康对照组、哮喘未干预组比较差异有显著性（Ｐ均＜０－０５） ;（４） 除哮喘ＮＴＧ     

哮喘患者外周血单个核细胞中白细胞介素—6mRNA表达与气道炎症的 …

目的 探讨外周血单个核细胞（ＰＢＭＣ）中白细胞介素（ＩＬ）－６ｍＲＮＡ表达与血嗜酸细胞阳离子蛋白（ＥＣＰ）和１秒用力呼气容积占用力肺活量比值（ＦＥＶ１％）的关系。方法 对１２例过敏性哮喘发作期患者、８例缓解期患者及９例健康人，采用ＲＴ－ＰＣＲ法和图像分析半定量法检测ＰＢＭＣ中ＩＬ－６ｍＲＮＡ的表达水平及ＥＣＰ和ＦＥＶ１％。结果 发作期患者ＩＬ－６ｍＲＮＡ的表达明显高于缓解期患者和健康人（Ｐ〈０．０     

SLE 患者 PBMC 分泌IL-3 活性水平的初步研究

目的：为了研究ＳＬＥ患者ＰＢＭＣ分泌ＩＬ３的能力。方法：采用ＭＴＴ比色方法，用ＩＬ３依赖株（ＴＦ１）分别测定了１５例活动期、１５例缓解期ＳＬＥ患者和正常对照者ＰＢＭＣ培养上清中ＩＬ３的活性水平。结果：ＳＬＥ患者ＰＢＭＣ自发分泌ＩＬ３的活性水平显著高于正常人（Ｐ＜０００１），活动期和缓解期患者之间无显著性差异（Ｐ＞００５），经ＰＨＡ刺激后ＳＬＥ患者和正常人ＰＢＭＣ分泌ＩＬ３的活性水平均显著增加（Ｐ＜０００１），在ＳＬＥ病人中，以伴发显著血小板减少的５例患者ＰＢＭＣ分泌ＩＬ３的活性水平最低。结论：ＩＬ３可能参与ＳＬＥ的致病过程，且和ＳＬＥ患者血小板减少有关。     

哮喘患者外周血单个核细胞中白细胞介素-6 mRNA表达与气道炎症的研究

目的探讨外周血单个核细胞（ＰＢＭＣ）中白细胞介素（ＩＬ）６ｍＲＮＡ表达与血嗜酸细胞阳离子蛋白（ＥＣＰ）和１秒用力呼气容积占用力肺活量比值（ＦＥＶ１％）的关系。方法对１２例过敏性哮喘发作期患者、８例缓解期患者及９例健康人，采用ＲＴＰＣＲ法和图像分析半定量法检测ＰＢＭＣ中ＩＬ６ｍＲＮＡ的表达水平及ＥＣＰ和ＦＥＶ１％。结果发作期患者ＩＬ６ｍＲＮＡ的表达明显高于缓解期患者和健康人（Ｐ＜００１），而缓解期患者无明显升高。发作期患者ＩＬ６ｍＲＮＡ的表达水平与血清ＥＣＰ呈中度正相关（ｒ＝０６７９，Ｐ＜００１），而与ＦＥＶ１％呈中度负相关（ｒ＝－０５８９，Ｐ＜００５）。结论过敏性哮喘发作期患者ＰＢＭＣ中ＩＬ６ｍＲＮＡ表达水平显著增强。其气道炎症程度与ＩＬ６基因转录增强有关     

SLE患者血清中白细胞介素-2受体和睾酮水平的变化

选择５９例系统性红斑狼疮（ＳＬＥ）患者和２０例正常人,检测白细胞介素－２受体（ｓＩＬ－２Ｒ）和睾酮（Ｔｃ）水平。用疾病活动评分（ＳＬＡＭ）判断疾病活动性,并对ｓＩＬ－２Ｒ和Ｔｃ水平的变化及两者回的相关性进行分析。结果ＳＬＥ患者血清ｓＩＬ－２Ｒ水平显著高于正常人（Ｐ〈０．００１）,Ｔｅ显著低于正常人（Ｐ〈０．００１）,ＳＬＥ活动期ｓＩＬ－２Ｒ水平与ＳＬＡＭ指数显著高于非活动期（Ｐ〈０．０１）,Ｔｅ显     

结缔组织病患者血清层粘连蛋白及Ⅳ型胶原测定的价值

目的为探讨层粘连蛋白（ＬＮ）、Ⅳ型胶原（Ⅳ·Ｃ）与活动期系统性红斑狼疮（ＳＬＥ）、系统性硬化症（ＳＳｃ）及皮肌炎（ＤＭ）的关系。方法采用放免方法对２０例活动期ＳＬＥ、６例ＳＳｃ、５例ＤＭ及２０例正常人血清ＬＮ及Ⅳ·Ｃ含量进行了测定。结果活动期ＳＬＥ及ＤＭ患者血清ＬＮ和Ⅳ·Ｃ水平明显升高（Ｐ＜００１）、ＳＳｃ组中Ⅳ·Ｃ明显升高（Ｐ＜００１）。结论ＬＮ、Ⅳ·Ｃ与ＡＮＡ、ｄｓＤＮＡ抗体及补体一起可能是评估ＳＬＥ活动的重要参数。它提示在ＳＬＥ、ＳＳｃ及ＤＭ的发展过程中ＬＮ和Ⅳ·Ｃ可能具有免疫致病性     

结缔组织病患者清层粘连蛋白及IV型胶原测定的价值

目的 为探讨层粘连蛋白（ＬＮ）、Ⅳ型胶原（Ⅳ·Ｃ）与活动期系统性红斑狼疮（ＳＬＥ）、系统性硬化症（ＳＳｃ）及皮肌炎（ＤＭ）的关系。方法 采用放免方法对２０例活动期ＳＬＥ、６例ＳＳｃ、５例ＤＭ及２０例正常人血清ＬＮ及Ⅳ·Ｃ明显升高（Ｐ〈０．０１）。结论 ＬＮ、Ⅳ·Ｃ与ＡＮＡ、ｄｓＤＮＡ抗体及补体一起可能是评估ＳＬＥ活动的重要参数。它提示在ＳＬＥ、ＳＳｃ及ＤＭ的发展过程中ＬＮ和Ⅳ·Ｃ可能具有免疫致病性     

系统性红斑狼疮患者外周血淋巴细胞早期凋亡的研究

目的：为了解观察ＳＬＥ患者休内细胞凋亡情况，并其外周血淋巴细胞早期凋亡率进行了检测，方法：应用最新的ＡｎｎｅｘｉｎＶ检测试剂盒及流式细胞仪检测早期凋亡细胞。结果：ＳＬＥ活动期患者外周血淋巴细胞早期凋亡率（１２．６１±６．３２，ｎ＝２）明显高于稳定期患者（４．４７±３．３９，ｎ＝８，Ｐ〈０．０１）及正常人（５．１３±３．３７，ｎ＝１１，Ｐ〈０．００１），但与ＳＬＡＭ评分无相关，活动期患者初发病组（１     

动脉平滑肌细胞一氧化氮合酶与高血压发病的关系

目的 探讨高血压病患者血管平滑肌细胞（ＶＳＭＣ）一氧化氮合酶（ＮＯＳ）活性和ＮＯＳ基因表达的特点及其与高血压发病的关系。方法 采用复合胶原酶法分离培养高血压病患者和血压正常者动脉ＶＳＭＣ,并以血红蛋白分光光度法、原位反转录聚合酶链反应检测ＶＳＭＣ的ＮＯＳ活性和诱生型ＮＯＳ（ｉＮＯＳ）ｍＲＮＡ表达量。结果 （１）高血压组ＶＳＭＣ的ＮＯＳ活性和ｉＮＯＳ ｍＲＮＡ表达量均显著低于正常组（Ｐ〈０．０１）;     

Elevation of plasma osteopontin concentration is correlated with disease activity in patients with systemic lupus erythematosus

BACKGROUND: Osteopontin (OPN) is an extracellular matrix cell adhesion phosphoprotein with immunological activities including stimulation of macrophage chemotaxis, T-helper type 1 lymphocyte response and B-cell antibody synthesis. Overexpression of OPN has been associated with the development of the autoimmune/lymphoproliferative syndrome. METHODS: We measured the plasma concentration and ex vivo production of OPN, and the plasma proinflammatory IL-18 concentration in 54 SLE patients with or without renal impairment (RSLE group and SLE group, respectively) and 26 sex- and age-matched control (NC) subjects using an enzyme-linked immunoabsorbent assay. RESULTS: Plasma OPN concentrations were significantly higher in RSLE and SLE patients than in the NC group (both P<0.001). Increase in OPN concentration correlated positively and significantly with SLEDAI score in all SLE patients (r = 0.308, P = 0.023). The ex vivo production of OPN upon mitogen activation of peripheral blood mononuclear cells was significantly higher in the RSLE and SLE groups than in the NC group (both P<0.001). In RSLE patients, plasma OPN concentration showed a significant positive correlation with proinflammatory cytokine IL-18 concentration (r = 0.404, P = 0.037). CONCLUSION: The above results suggest that the production of OPN is associated with the inflammatory process and SLE development, and may serve as a potential disease marker of SLE.     

Overexpression of osteopontin in rheumatoid synovial mononuclear cells is associated with joint inflammation, not with genetic polymorphism

OBJECTIVE: Osteopontin (OPN) is thought to play an important role in rheumatoid synovitis. We investigated the expression of OPN in rheumatoid synovial fluid mononuclear cells (SFMC) and its potential association with genetic polymorphism of the OPN gene and joint inflammation in rheumatoid arthritis (RA). METHODS: 1. The expression of OPN mRNA in peripheral blood mononuclear cells (PBMC) and SFMC of patients with RA was analyzed quantitatively by real-time polymerase chain reaction (PCR). Results were analyzed in paired PBMC and SFMC and control PBMC. 2. Six single nuclear acid polymorphisms of the OPN gene were genotyped in a cohort of 192 Chinese patients with RA and controls (n = 288) by restriction fragment length polymorphism PCR or direct DNA sequencing. 3. SF derived from RA patients was examined for the stimulating effect on mRNA expression of the OPN gene in PBMC. RESULTS: The expression of OPN gene was significantly increased in SFMC and, to a lesser degree, in PBMC of patients with RA compared to control PBMC (p < 0.01). However, the prevalence of OPN genotype and allele frequencies at the selected positions did not differ significantly between RA patients and the control group (p > 0.05). Further characterization indicated that SF known to contain a variety of proinflammatory factors significantly stimulated mRNA expression of OPN in PBMC obtained from RA patients or healthy controls. CONCLUSION: Overexpression of OPN mRNA in SFMC is associated with proinflammatory factors produced in inflamed joints, but not with OPN genetic polymorphisms. OPN gene polymorphisms do not correlate with susceptibility to RA.     

Dysregulated expression of interleukin-23 and interleukin-12 subunits in systemic lupus erythematosus patients

The aim of this study was to investigate the regulation of interleukin (IL)-12 and IL-23 expression in the autoimmune disease, systemic lupus erythematosus (SLE). mRNA from healthy subjects and SLE patients were prepared from peripheral blood mononuclear cells (PBMC) and quantitative real-time polymerase chain reaction was performed to quantify IL-23 specific subunit P19, IL-12 specific subunit P35, and their common subunit P40. IL-12 specific subunit P35 mRNA expression in untreated and treated SLE patients was significantly lower than healthy controls (P = 0.015 and 0.000, respectively). Compared with untreated SLE patients, treatment of SLE patients with corticosteroids or corticosteroids plus another immunosuppressor significantly suppressed P40 and P19 expression (P = 0.002 and 0.015, respectively). The mRNA levels of p19, p40, and p35 in active SLE patients (SLEDAT > 10) were significantly higher compared with those in the inactive SLE patients (SLEDAI ≤ 10) (P = 0.000, 0.000, and 0.017, respectively). These results suggest that deficiency of IL-12 and possibly upregulation of IL-23 may contribute to SLE pathogenesis and both cytokines may be therapeutic targets in SLE.     

外周血单个核细胞巨噬细胞移动抑制因子表达及其与狼疮活动

目的 观察系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)巨噬细胞移动抑制因子(MIF)mRNA表达及血MIF水平的变化,并分析其与SLE疾病活动性、血清免疫学指标的相关关系。方法 采用逆转录-PCR技术及ELISA方法分别测定34例活动期和21例静止期SLE患者的PBMC中MIF mRNA表达及血清MIF浓度,并以18例正常人作对照,观察SLE患者血MIF的改变及其与疾病活动性、血清免疫学指标的相关关系。结果 SLE患者PBMC中MIF mRNA表达及血MIF水平较正常人显著升高(P<0.01),且活动期患者增高更为明显,显著高于静止期患者(P<0.01)。SLE患者血MIF水平及PBMC中MIF mRNA表达与疾病活动指数、抗ds-DNA抗体、血补体C3、C4及IgG水平明显相关,但与抗核抗体无关。结论 SLE患者血MIF mRNA表达及血MIF水平显著增高,其变化可反映SLE患者疾病的活动性。     

系统性红斑狼疮患者外周血单个核细胞环氧合酶同工酶mRNA的表达

目的　探讨原生型环氧合酶 (COX 1)和诱生型环氧合酶 (COX 2 )在系统性红斑狼疮(SLE)患者外周血单个核细胞 (PBMC)中的表达水平及其在SLE发病机制中的作用及其意义。方法　应用逆转录 聚合酶链反应 (RT PCR)检测了 34例活动期SLE患者和 30名正常人PBMC中COX 1和COX 2mRNA的表达水平。结果　活动期SLE患者COX 1和COX 2的阳性表达率与正常对照组相比差异无显著性 (P >0 0 5 ) ;活动期SLE患者PBMC中COX 2mRNA的平均表达水平 (0 6 6±0 2 7)明显高于正常对照组 (0 43± 0 16 ) ,差异有非常显著性 (P <0 0 1) ;活动期SLE组COX 1的平均表达水平与正常对照组处于同一表达水平 ,差异性无统计学意义 (P >0 0 5 )。结论　活动期SLE患者外周血单个核细胞中COX 2mRNA的表达增加 ,通过诱导前列腺素 (PGs)的产生参与SLE的发病过程。为特异性COX 2抑制剂应用于SLE的治疗提供了理论依据。     

系统性红斑狼疮患者血浆骨桥蛋白水平与疾病活动性及脏器损害的相关性研究

目的 检测系统性红斑狼疮(SLE)患者血浆中骨桥蛋白(OPN)的水平,分析其与SLE活动性及脏器损害间的关系.方法 选择68例SLE患者和36名健康体检者,采用酶联免疫吸附试验(ELISA)检测其血浆中的OPN水平.记录采血时患者的系统性红斑狼疮疾病活动指数(SLEDAI)、实验室指标等.结果 SLE患者OPN血浆水平明显高于正常对照组[(4.5±2.0)ng/ml与(1.6±0.7)ng/ml,P＜0.01 ].SLE疾病活动组高于非活动组[(5.3±2.0)ng/ml与(3.4±1.3)ng/ml,P＜0.01];SLE患者伴有肾脏损害组高于无肾脏损害组[(5.8±2.1)ng/ml与(3.5±1.3)ng/ml,P＜0.01],有肺间质病变、胃肠道病变和心包炎的高于无脏器损害[(4.8±1.2)、(6.3±1.4)、(5.4±2.6)ng/ml与(3.5+1.3)ng/ml,P＜0.05],抗心磷脂抗体阳性患者OPN水平高于抗心磷脂抗体阴性患者[(5.3±2.4)ng/ml与(4.2±1.7)ng/ml,P＜0.05].OPN血浆水平与SLEDAI呈正相关(r=0.523,P＜0.01),与尿蛋白定量(24 h)呈正相关r=0.403,P=0.001),与补体C3呈负相关r=-0.398,P=0.001),与红细胞沉降率(ESR)、抗dsDNA抗体、抗Sm抗体、IgG、IgA、IgM、球蛋白及关节炎等无相关性(P＞0.05) .结论 OPN可能参与SLE的发病,临床上可作为疾病活动及脏器损伤的观察指标.     

The effect of hyaluronic acid on osteopontin and CD44 mRNA of fibroblast-like synoviocytes in patients with osteoarthritis of the knee

The aim of this study was to investigate the effect of hyaluronic acid (HA) on the expression of osteopontin (OPN) and CD44 mRNA of fibroblast-like synoviocytes (FLS) in the patients with osteoarthritis (OA) of the knee. FLS were obtained from the knees of 10 patients with OA. Real-time quantitative polymerase chain reaction (Q-PCR) was used to assess the expression of the OPN mRNA and CD44 mRNA. The relative OPN mRNA expression in HA group (6.47 ± 2.30-fold) was significantly higher than in the control group (P = 0.045, P < 0.05), while in HYL group (0.65 ± 0.21-fold) it was lower than in the control group (P = 0.037, P < 0.05), and the difference in OPN mRNA expression between HA group and HYL group also showed statistically significant (P = 0.001, P < 0.05); however, there was no significant difference between each group of the relative CD44 mRNA expression (P > 0.05). Our study showed that HA can upregulate OPN mRNA expression in OA fibroblast-like synoviocytes, and the high expression of OPN mRNA in OA may be a result of increased HA level of OA synovitis; however, HA cannot affect the CD44 mRNA expression in OA fibroblast-like synoviocytes, and the high expression of CD44 mRNA in OA may be not a result of increased HA level of OA synovitis.     

淋巴细胞趋化因子受体mRNA在系统性红斑狼疮患者外周血单个核细胞的表达

目的探讨淋巴细胞趋化因子受体(XCR1)mRNA在系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的表达,及其与疾病的相关性。方法收集93例确诊的SLE患者和30名健康对照组,收集PBMC,提取RNA。应用反转录-聚合酶链反应(RT-PCR)方法检测研究对象XCR1mRNA表达水平。结果!"XCR1mRNA在PBMC中表达水平,患者组(1.57±0.84)与正常对照组(0.19±0.14),两者差异无统计学意义(P>0.05)。活动期(2.09±1.38)与对照组比较,两者差异有统计学意义(t=2.392,P=0.02);活动期与非活动期(0.31±0.24)比较,差异有统计学意义(t=3.091,P=0.003);非活动期与对照组比较,差异无统计学意义(P>0.05)。"#SLE患者组XCR1mRNA水平与疾病活动度评分(SLEDAI)关系:SLE患者组PBMC中XCR1mRNA表达水平与SLEDAI作直线相关性分析,XCR1mRNA水平与SLEDAI(r=0.540,t=5.445,P=0.000)呈正相关。"$SLE患者PBMC中XCR1mRNA表达与临床表现及实验室检查相关性:SLE抗SSA抗体阳性患者(17/68,1.4±1.2)比阴性患者(51/68,3.5±4.9),其XCR1mRNA表达水平降低,差异有统计学意义(t=2.78,P=0.007)。结论XCR1mRNA表达水平在PBMC中活动期SLE患者比非活动期及对照组增高,与疾病的活动性(SLEDAI)正相关,非活动期与对照组差异无统计学意义。SLE抗SSA抗体阳性患者比阴性患者,其XCR1mRNA表达水平降低,两组均较健康对照组增高。提示XCR1参与疾病的发展,与疾病的活动性相关,XCR1可能参与抗SSA抗体的形成过程,造成组织损伤。     

SLE患者外周血CD4+ CD25+调节性T细胞及相关基因Foxp3的变化

目的研究系统性红斑狼疮（SLE）合并狼疮性肾炎患者经肾上腺糖皮质激素（简称激素）冲击治疗后，外周血CD4^＋CD25^＋调节性T细胞（regulatoryTcell，Treg）及相关基因Foxp3表达的变化，从而探讨CD4^＋CD25^＋Treg和Foxp3与SLE发病的相关性。方法采用流式细胞术检测SLE患者外周血单个核细胞（PBMC）中CD4^＋CD25^＋T、CD4^＋CD25highT细胞数量的变化，RTPCR检测PBMC中CD4^＋CD25^＋Treg功能相关基因Foxp3mRNA的表达。结果激素冲击治疗后的SLE患者CD4^＋CD25^＋T／CD4^＋T及CD4^＋CD25highT／CD4^＋T比值高于正常对照组；Foxp3mRNA水平表达与对照组差异不显著。结论CD4^＋CD25^＋Treg数量和功能的变化可能参与SLE的发病，激素可能通过提升CD4^＋CD25^＋Treg治疗SLE。     

骨桥蛋白在系统性红斑狼疮中的临床意义

目的 研究血清骨桥蛋白(OPN)与系统性红斑狼疮(SLE)临床表现、实验指标、疾病活动的相关性,探讨血清OPN在SLE中的意义及SLE的发病机制.方法 用酶联免疫吸附(ELISA)法检测68例SLE患者和20名正常对照组血清中OPN的水平,并监测SLE患者的临床表现及实验室指标,分析其与OPN的相关性.结果 68例SLE患者中79%血清OPN阳性,20名正常对照组0PN均阴性,SLE组患者血清OPN阳性率及OPN水平显著高于正常对照组(P＜0.01).血清OPN阳性组的SLE患者与OPN阴性组的SLE患者相比,在年龄、性别及病程上差异均无统计学意义(P＞O.05 .血清OPN阳性组SLE患者发热、脱发、白细胞降低、肝脏损害、补体C4降低、免疫球蛋白IgA增高、蛋白尿及抗dsDNA抗体阳性的发生率明显高于OPN阴性组(P＜0.05). SLE患者血清OPN阳性率明显高于抗dsDNA抗体、抗sm抗体的阳性率,差异具有统计学意义(P＜0.01). SLE患者血清OPN水平与SLE疾病活动指数(SLEDAI)积分呈明显的正相关(r=0.292;P＜0.05);SLE活动期患者血清OPN阳性率及水平明显高于SLE缓解期患者,两组比较差异具有统计学意义(P＜O.05).结论 SLE患者血清中OPN水平升高,与SLE疾病活动性密切相关,可作为SLE的病情活动指标.