转化生长因子β1对长波紫外线照射皮肤成纤维细胞线粒体DNA 4 977 bp缺失的影响

背景:长波紫外线与人体皮肤光老化关系密切,线粒体的损伤是细胞衰老和死亡的分子基础.目的:观察长波紫外线照射对体外培养的皮肤成纤维细胞的线粒体脱氧核糖核酸缺失损伤的影响,以及转化生长因子β1对长波紫外线引起的线粒体DNA缺失有无保护作用.为皮肤光老化研究提供实验依据.设计、时间及地点:实验于2007-03/2008-04于解放军第四军医大学西京医院全军整形外科研究所完成.材料:转化生长因子β1为PerProtech公司产品;线粒体DNA 4 977 bp引物由上海生工合成;长波紫外线光源为北京光学仪器厂生产:紫外线辐照计为北京师范大学光电仪器厂生产.方法:收集20-23岁成年男性包皮环切术后的皮肤组织12例,体外培养成纤维细胞.将细胞分组为对照组、长波紫外线累计照射达到30,60,90 J,cm~2组,半定量PCR检测DNA4 977 bp缺失情况.不同质量浓度转化生长因子β1(0.1,1,1 0μg/L)干预长波紫外线累积照射达90 J/cm~2的皮肤成纤维细胞.主要观察指标:观察不同累积照射剂量产生的DNA 4 977 bp缺失:观察累积照射剂量90 J/cm~2后不同浓度转化生长因子干预对DNA4 977 bp缺失的影响.结果:体外培养的皮肤成纤维细胞经长波紫外线照射,长波紫外线累积剂量为长波紫外线60 J/cm~2后发生线粒体DNA4977 bp缺失,长波紫外线90 J/cm~2时缺失加重.吸光度值和PCR产物电泳及条带密度扫描结果显示,在照射前2 h加入不同剂量转化生长因子处理后,大剂量组(10 μg/L)线粒体DNA表达降低,中、小剂量组与长波紫外线照射组比较,差异无显著性意义.结论:一定剂量(10μg/L)转化生长因子对体外培养的成纤维细胞线粒体DNA缺失起到保护作用.     

转化生长因子β1对长波紫外线照射皮肤成纤维细胞线粒体DNA 4977 bp缺失的影响

背景：长波紫外线与人体皮肤光老化关系密切,线粒体的损伤是细胞衰老和死亡的分子基础。目的：观察长波紫外线照射对体外培养的皮肤成纤维细胞的线粒体脱氧核糖核酸缺失损伤的影响,以及转化生长因子β1对长波紫外线引起的线粒体DNA缺失有无保护作用。为皮肤光老化研究提供实验依据。设计、时间及地点：实验于2007-03/2008-04于解放军第四军医大学西京医院全军整形外科研究所完成。材料：转化生长因子β1为 PerProtech公司产品;线粒体DNA 4 977 bp引物由上海生工合成;长波紫外线光源为北京光学仪器厂生产;紫外线辐照计为北京师范大学光电仪器厂生产。 方法：收集20~23岁成年男性包皮环切术后的皮肤组织12 例,体外培养成纤维细胞。将细胞分组为对照组、长波紫外线累计照射达到30,60,90 J/cm2组,半定量PCR检测DNA 4 977 bp缺失情况。不同质量浓度转化生长因子β1（0.1,1,10 μg/L）干预长波紫外线累积照射达90 J/cm2的皮肤成纤维细胞。主要观察指标：观察不同累积照射剂量产生的DNA 4 977 bp缺失;观察累积照射剂量90 J/cm2后不同浓度转化生长因子干预对DNA4 977 bp缺失的影响。结果：体外培养的皮肤成纤维细胞经长波紫外线照射,长波紫外线累积剂量为长波紫外线60 J/cm2后发生线粒体DNA 4 977 bp 缺失,长波紫外线90 J/cm2时缺失加重。吸光度值和PCR产物电泳及条带密度扫描结果显示,在照射前2 h加入不同剂量转化生长因子处理后,大剂量组（10 μg/L）线粒体DNA表达降低,中、小剂量组与长波紫外线照射组比较,差异无显著性意义。结论：一定剂量（10 μg/L）转化生长因子对体外培养的成纤维细胞线粒体DNA缺失起到保护作用。     

线粒体DNA与细胞癌变关系的研究

研究表明 ,外源性遗传物质如 DNA病毒或 RNA病毒在细胞基因组中整合能诱导细胞恶性转化。但作为哺乳动物细胞唯一核外基因组的线粒体 DNA( mt DNA) ,是否也具有类似于肿瘤病毒那样的活性呢 ?关于此问题的研究已从理论上进行了大胆的推测 ,认为线粒体这一最容易受损伤的细胞器 ,它所释放的遗传物质有可能具有潜在的致癌性。但此观点尚缺乏充分的实验证据。为了探讨 mt DNA与细胞癌变的关系 ,本实验采用地高辛 ( DIG)标记的人 mt D-NA探针 ,对 6株人癌细胞系和 1例原代培养人皮肤成纤维细胞的染色体或间期细胞核进行了荧光原位杂交( …     

银杏叶提取物对老年大鼠脑组织线粒体DNA缺失的影响

目的：测定灌服银杏叶提取物后老年大鼠大脑皮质、海马和小脑线粒体DNA缺失情况，为研究银杏治疗老年痴呆的分子机制提供基础资料。方法：用碱裂解法抽提灌服银杏叶提取物的老年大鼠15只，对照组15只的大脑皮质、海马和小脑的mtDNA。PCR法扩增mtDNA部分片段，检测mtDNA缺失片段并测定其A值。结果：大鼠各脑区均有缺失型的mtDNA，缺失片段为4800bp，灌服银杏叶提取物的老年大鼠各脑区的缺失型mtDNA的光密度值：大脑皮质为0.29，海马为0．27，远远小于对照组大鼠各脑区的缺失型mtDNA的A值：大脑皮质为0．59，海马为0．53。结论：灌服银杏叶提取物的大鼠各脑区缺失型mtDNA的量明显减少，提示银杏叶提取物可能通过抑制氧自由基对线粒体DNA的损害而延缓老年大鼠脑衰老的进程。     

人肿瘤中线粒体DNA的缺陷

线粒体DNA(mtDNA)是母系遗传的胞浆DNA,以高拷贝存在于细胞中。自身结构特点决定其在肿瘤中的突变率显著高于核DNA(nDNA),所以研究者们认为mtDNA的缺陷可能促进肿瘤的发生和发展。该文就近年来肿瘤中mtDNA的常见缺陷作一综述,以探讨mtDNA缺陷与肿瘤的关系,为临床诊断和治疗肿瘤提供参考。     

缺失线粒体DNA的人喉癌细胞系的建立

目的为了研究线粒体DNA在人喉癌细胞的增殖、分化及凋亡中的影响及作用,培养线粒体DNA缺失的喉癌细胞系,为构建融合喉癌细胞,研究线粒体DNA突变和线粒体功能改变与喉癌发生之间的关系奠定基础.方法人喉癌细胞系JHUo11加入10%FBS的RPMI1640培养基中,然后在培养基中加入丙酮酸、尿苷、溴化乙锭(EtBr)以去除线粒体DNA,不加溴化乙锭的o11细胞作为对照.经过90天培养,线粒体DNA缺失的喉癌细胞通过健那绿染色后光镜观察和PCR法鉴定.结果 o11细胞经过75ng/μl溴化乙锭持续作用90天后可见细胞肿胀变圆,透光度增加,线粒体DNAPCR扩增未见目的条带,细胞可扩增出定位于mtDNA的500bp片段,健那绿染色对照细胞,可见散在蓝绿色线粒体,而加EB的细胞未见线粒体.结论喉癌细胞o11经过溴化乙锭持续作用90天培养出线粒体DNA缺失的ρ0o11喉癌细胞,这种缺失线粒体DNA的细胞系需补充外源性丙酮酸和尿苷来维持其生存,否则细胞很快死亡.通过观察细胞的增殖和分化发现,ρ0o11细胞生长速度比未经EB处理的对照组慢的多.     

急性淋巴细胞白血病线粒体DNA突变的研究

目的:研究急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)患者白细胞线粒体DNA D-loop区突变情况,以探讨线粒体DNA突变与急性白血病发生发展的关系。方法:采用聚合酶链反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)和DNA序列分析相结合的方法,对15例ALL病人治疗前、缓解期及复发时白细胞线粒体基因组的D-loop片段进行突变研究。结果:以缓解期为对照,发现8例(54%)ALL病人治疗前白细胞的线粒体DNA D-loop片段上出现突变,复发时又有相同或不同的突变出现;突变类型均为碱基替代突变,主要是C—T突变(43%);同时发现在线粒体DNA的某些位点发生突变的频率明显较高。结论:白细胞线粒体基因组的突变可能在白血病的发生发展过程中起重要作用。     

自由基氧化致线粒体DNA损伤与细胞凋亡

细胞凋亡与自由基氧化密切相关,自由基氧化可致线粒体DNA损伤而发生突变,引起细胞裹老甚至死亡;细胞死亡包括坏死和调亡,线粒体DNA损伤突变与细胞凋亡间有无关系？本文就此对自由基氧化、线粒体DNA损伤以及细胞凋亡相关基因及其与细胞凋亡之间的关系作一综述。     

银杏叶提取物对老年大鼠脑组织线粒体DNA缺失的影响

目的:测定灌服银杏叶提取物后老年大鼠大脑皮质、海马和小脑线粒体DNA缺失情况,为研究银杏治疗老年痴呆的分子机制提供基础资料。方法:用碱裂解法抽提灌服银杏叶提取物的老年大鼠15只,对照组15只的大脑皮质、海马和小脑的mtDNA。PCR法扩增mtDNA部分片段,检测mtDNA缺失片段并测定其A值。结果:大鼠各脑区均有缺失型的mtDNA,缺失片段为4800bp,灌服银杏叶提取物的老年大鼠各脑区的缺失型mtDNA的光密度值:大脑皮质为0.29,海马为0.27,远远小于对照组大鼠各脑区的缺失型mtDNA的A值:大脑皮质为0.59,海马为0.53。结论:灌服银杏叶提取物的大鼠各脑区缺失型mtDNA的量明显减少,提示银杏叶提取物可能通过抑制氧自由基对线粒体DNA的损害而延缓老年大鼠脑衰老的进程。     

衰老小鼠脑老化过程中线粒体结构功能与mtDNA缺失的生物学规律

目的:研究小鼠脑老化过程中线粒体的生物学变化规律,探讨衰老的机制。方法:实验于2002-01/2003-04在广州体育学院科学实验中心及中山大学医学院完成。利用电镜对线粒体进行观察计数,Clark氧电极法测定线粒体呼吸链细胞色素C氧化酶及NADH脱氢酶活性,分光光度法测定抗氧化酶活性,聚合酶链反应检测mtDNA3866bp片段缺失率。结果:与5月龄小鼠比较,20月龄的老年小鼠脑线粒体数量减少、体积增大,线粒体呼吸控制率减小,而ADP/O比值无显著性差异,呼吸链NADH脱氢酶、细胞色素C氧化酶活性下降,mtDNA3866bp片段缺失率增加,而抗氧化酶活性却增大。通过各指标间的相关分析发现呼吸控制率与mtDNA缺失率显著相关(r=0.739,P<0.01),NADH脱氢酶、细胞色素C氧化酶活性与mtDNA缺失率亦显著相关(r=0.582,P<0.05,r=0.810,P<0.01),但抗氧化酶活性与mtDNA缺失率无相关性(r=0.256,P>0.05)。结论:推测衰老时mtDNA的损伤积累可引起呼吸链酶复合物活性下降,导致呼吸链功能减退致生物衰老。     

短波紫外线照射后大鼠皮肤羟脯氨酸含量的变化

目的观察大鼠皮肤伤口受不同剂量短波紫外线照射后组织中羟脯氨酸含量的变化 ,探讨紫外线照射对胶原蛋白合成的影响。方法用 3 0只Wistar雄性大鼠建立短波紫外线照射新鲜伤口的动物模型 ,在每只大鼠颈背部做 3个直径 2cm的圆形皮肤全层伤口 ,其中 2个分别接受为 15MED( 15mJ/cm2 )和 60MED( 60mJ/cm2 )短波紫外线照射 ,另 1个作为对照不接受照射 ,然后采用化学法检测伤口肉芽组织中羟脯氨酸含量的变化。结果 15MED照射伤口在照射后 2 1— 2 8d时羟脯氨酸含量高于对照 (P <0 .0 5 ) ;60MED照射伤口在 2 8d时羟脯氨酸含量显著增高 ,与 15MED照射伤口和对照之间的差异具有高度显著性意义 (P <0 .0 1)。结论短波紫外线照射能促进伤口中羟脯氨酸的合成 ,从而增加胶原含量 ,促进伤口愈合 ;60MED短波紫外线照射的作用强于 15MED。     

Massive thymic deletion results in systemic autoimmunity through elimination of CD4+ CD25+ T regulatory cells

Incomplete deletion of KRN T cells that recognize the ubiquitously expressed self-antigen glucose-6-phosphate-isomerase (GPI) initiates an anti-GPI autoimmune cascade in K/BxN mice resulting in a humorally mediated arthritis. Transgenic (Tg) expression of a KRN T cell receptor (TCR) agonist under the major histocompatibility complex class II promoter resulted in thymic deletion with loss of anti-GPI T and B cell responses and attenuated arthritis course. However, double Tg mice succumbed to systemic autoimmunity with multiorgan inflammation and autoantibody production. Extensive thymic deletion resulted in lymphopenia and elimination of CD4+ CD25+ regulatory T cells (Tregs), but spared some CD4+ T cells expressing endogenous TCR, which oligoclonally expanded in the periphery. Disease was transferred by these T cells and prevented by cotransfer of CD4+ CD25+ Tregs. Moreover, we extended our findings to another TCR system (anti-hen egg lysozyme [HEL] TCR/HEL mice) where similarly extensive thymic deletion also resulted in disease. Thus, our studies demonstrated that central tolerance can paradoxically result in systemic autoimmunity through differential susceptibility of Tregs and autoreactive T cells to thymic deletion. Therefore, too little or too much negative selection to a self-antigen can result in systemic autoimmunity and disease.     

鱼藤酮诱导PC12细胞损伤作用途径

目的探讨鱼藤酮诱导PC12细胞损伤作用的可能途径，为神经防护药物的研发提供理论基础。方法将培养的PC12细胞分为正常对照组和0．5μmol&#183;L^-1鱼藤酮实验组，持续作用72h后MTT法检测细胞存活率、Hoechst 33342观测细胞凋亡、提取实验组和对照组细胞的总蛋白，应用荧光差异凝胶电泳系统（Differential Gel Electrophoresis，DIGE）获得差异蛋白点的表达信息，通过MALDI-TOF质谱鉴定差异蛋白点。结果MTT法显示鱼藤酮实验组较正常对照组细胞存活率明显降低（P〈0．01）；Hoechst33342荧光染色显示鱼藤酮实验组凋亡率明显高于对照组（P〈0．01）.DIGE分析软件提示实验组与对照组相比发现三个有意义的差异蛋白。通过质谱分析和数据库检索，鉴定分别为γ-烯醇化酶（γ-enolase）、磷酸丙糖异构酶1（Trpi1）和真核细胞翻译起始因子4A（elF4A）。结论γ-enolase、Tpi1和elF4A参与细胞损伤。可能成为神经防护药物作用的靶点。     

CD4+ CD25+ regulatory T cells control T helper cell type 1 responses to foreign antigens induced by mature dendritic cells in vivo

Recent evidence suggests that in addition to their well known stimulatory properties, dendritic cells (DCs) may play a major role in peripheral tolerance. It is still unclear whether a distinct subtype or activation status of DC exists that promotes the differentiation of suppressor rather than effector T cells from naive precursors. In this work, we tested whether the naturally occurring CD4+ CD25+ regulatory T cells (Treg) may control immune responses induced by DCs in vivo. We characterized the immune response induced by adoptive transfer of antigen-pulsed mature DCs into mice depleted or not of CD25+ cells. We found that the development of major histocompatibility complex class I and II-restricted interferon gamma-producing cells was consistently enhanced in the absence of Treg. By contrast, T helper cell (Th)2 priming was down-regulated in the same conditions. This regulation was independent of interleukin 10 production by DCs. Of note, splenic DCs incubated in vitro with Toll-like receptor ligands (lipopolysaccharide or CpG) activated immune responses that remained sensitive to Treg function. Our data further show that mature DCs induced higher cytotoxic activity in CD25-depleted recipients as compared with untreated hosts. We conclude that Treg naturally exert a negative feedback mechanism on Th1-type responses induced by mature DCs in vivo.     

电离辐射诱导HepG2细胞中Nucleolin和p53变化时程研究

目的检测电离辐射照射HepG2细胞后,其Nucleolin和p53蛋白的变化情况,结果将为进一步揭示电离辐射对p53信号传导通路的影响提供重要的细胞学基础,并为辐射生物效应研究提供新的生物学依据。方法电离辐射照射HepG2细胞后,分别提取总蛋白、胞浆蛋白和胞核蛋白,然后采用流式和WesternBlot的方法进行蛋白检测。结果HepG2细胞接受4Gy电离辐射照射后,其总蛋白中的Nucleolin和p53表达水平增高,且与照射时间呈正相关;HepG2细胞接受电离辐射照射后的2h、4h和8h胞核蛋白中的Nucleolin和p53表达水平下降,而胞浆蛋白中的Nucleoiin和p53表达水平对应增多。结论HepG2细胞受到一定剂量的电离辐射刺激后,Nucleolin和p53在肿瘤细胞内有明显的由胞核至胞浆的穿梭作用,二者的穿梭具有一定的协同性。     

三种不同方法体外诱导CD4+T细胞向Th17细胞分化与增殖的效率

背景:研究认为Th17细胞参与了许多炎症性疾病的发生,但其体外诱导分化效率尚不明确.目的:比较采用3种不同方法在体外诱导CD4+T细胞向Th17细胞分化与增殖的效率.方法:分别采取以下3种方法体外诱导Th17细胞:①给予CD3及CD28抗体刺激.并在培养体系中加入白细胞介素6及转化生长因子β.培养3d.②在方法①的基础上加入白细胞介素1β及肿瘤坏死因子α,培养3d.③在方法②的基础上第3天洗去细胞因子,培养2d;再次给予CD3及CD28抗体刺激,并在培养体系中加入白细胞介素23,培养3d.结果与结论:3种方法培养后CD4+T细胞中Th17细胞的比例分别为(8.5±2.8)%,(26.9±4.3)%,(44.3±5.5)%,三组之间差异有显著性意义(P<0.01).提示在培养体系中加入白细胞介素1β、肿瘤坏死因子α及白细胞介素23,更有利于增加CD4+T细胞向Th17细胞分化的比例.     

Intrathymic clonal deletion of V beta 6+ T cells in cyclophosphamide-induced tolerance to H-2-compatible, Mls-disparate antigens

When C3H (H-2k, Mls-1b) mice were primed intravenously with 10(8) viable spleen cells from AKR (H-2k, Mls-1a) and treated intraperitoneally with 200 mg/kg of cyclophosphamide (CP) 2 d later, not only a long-lasting skin allograft tolerance but also a tolerance in mixed lymphocyte reaction to Mls-1a-encoded antigens was established. The cellular mechanisms of CP-induced tolerance were examined by assessing the V beta 6-bearing T cells that are strongly correlated with reactivity to Mls-1a-encoded antigens bound to MHC class II molecules. At the relatively early stage (2 or 5 wk) after the CP treatment, CD4+-V beta 6+ T cells of C3H origin were preferentially eliminated in the lymph nodes of the tolerant mice, whereas CD8+-V beta 6+ T cells remained. On the other hand, neither CD4+CD8- nor CD4-CD8+ thymocytes bearing a high density of V beta 6 was detected in the chimeric thymus. Namely, in the thymus of the tolerant C3H mice, neither mixed chimerism nor the clonal deletion of the V beta 6-bearing T cells was observed on day 14, whereas both of them were observed on day 35. The clonal deletion and mixed chimerism in the thymus were lasting for greater than 10 wk after the CP treatment. Expression of V beta 6 on the peripheral T cells in the tolerant C3H mice gradually reduced in the process of time. These results strongly suggested that the clonal deletion in the thymus was one of the essential mechanisms in the CP-induced tolerance system.     

CD4+CD25+ regulatory T cells that secrete TGFbeta and IL-10 are preferentially induced by a vaccine vector

CD4CD25 T cells generated in a vaccine scenario can play a critical role in limiting antitumor therapy, thus having widespread implications for the immunotherapy-based treatment of cancer. The authors previously used Listeria monocytogenes to develop two vaccine constructs that express HPV-16 E7 protein and induce strong cellular immunity to HPV-E7-expressing tumors. Immunization of mice bearing established E7-expressing tumors with Lm-LLO-E7 induced regression of the tumors, whereas Lm-E7 showed little or no tumor regression. To investigate the possibility that regulatory CD4CD25 T-cell populations may be responsible for the differences in tumor regression, the authors characterized the role of these cells generated by the two vaccine systems. The authors compared the prevalence of CD4CD25 T cells in tumor-bearing vaccinated mice and demonstrate that Lm-E7-vaccinated mice have significantly increased numbers of CD4CD25 T cells in both the spleen and tumor-infiltrating lymphocytes compared with Lm-LLO-E7-vaccinated mice. The authors confirm that these increased numbers of CD4CD25 T cells are indeed suppressor in function by in vitro suppression assays and that the mechanism of action of the tumor-infiltrating cells involves the production of suppressor cytokines interleukin-10 and transforming growth factor beta. These results show that it is possible for a tumor vaccine system to generate tumor-infiltrating CD4CD25 regulatory T cells that critically affect tumor regression and the overall success of vaccine therapy.     

镓铝砷激光局部治疗对类风湿关节炎患者调节T细胞和局部炎性介质的影响

目的 探讨镓铝砷激光联合药物治疗类风湿关节炎(RA)的机制.方法 将22例RA患者分为观察组和对照组,每组11例.观察组采用镓铝砷激光+甲氨蝶呤治疗,对照组仅采用甲氨蝶呤治疗.采用流式细胞双抗染色法分别测定患者治疗前、后外周血和膝关节滑液中CD4+CD25+treg(调节性T细胞)的数量,采用酶联免疫吸附法测定治疗前、后患者膝关节滑液中前列腺素E2(PGE2)的含量.同时检测10名健康人(正常对照组)外周血CD4+CD25+treg细胞数量.结果 观察组膝关节局部症状改善优于对照组;2组治疗后膝关节滑液中PGE2含量分别为(3.82±1.34)和(1.69±0.98),均较治疗前下降,其中观察组下降幅度明显高于对照组,差异具有统计学意义(P＜0.05);患者外周血CD4+CD25+treg细胞数量为(3.84±3.20)%,明显低于正常对照组,差异存在统计学意义(P＜0.05).滑液中CD4+CD25+treg远多于外周血.观察组治疗后关节滑液中CD4+CD25+treg的数量为(9.78±10.28)%,与治疗前比较差异存在统计学意义(P＜0.05).结论 镓铝砷激光联合甲氨蝶呤治疗类风湿关节炎临床疗效优于单纯应用甲氨蝶呤,联合治疗的机制可能在于明显减少靶器官局部炎性介质和CD4+CD25+treg细胞数量.

Abstract：

Objective To explore the mechanism of combined treatment with methotrexate (MTX) and Ga-Al-As laser irradiation for rheumatoid arthritis (RA) and to assess the effectiveness of Ga-Al-As laser therapy for RA. Methods Twenty-two patients with RA were randomly and evenly divided into two groups: the treatment group treated with Ga-Al-As laser irradiation combined with MTX and the control group treated with MTX only. Ten age-matched normal subjects were observed as normal controls. The amount of CD4 + CD25 + regulatory T cells in peripheral blood (PB) of the normal controls and that in the PB and synovial fluid (SF) of the 22 patients before and after therapy were counted by flow cytometry. Meanwhile, the amount of prostaglandin E2 (PGE2) in synovial fluid of the patients was measured before and after treatment by enzyme-linked immunosorbent assay(ELISA). Results After combined treatment the clinical symptoms of the patients were improved significantly, and the amount of PGE2in SF decreased significantly. The count of CD4 + CD25 + regulatory T cells in PB of RA patients was ( 3.84 ±3.20) % , compared to ( 10.05 ± 7.04) % in healthy individuals. The count of CD4 + CD25 + regulatory T cells in SF of RA patients was ( 14.89 ± 12.30) % , much higher than that in PB. The count of CD4 + CD25 + regulatory T cells in SF decreased significantly in treatment group compared to control group (P ＜0.05). Conclusion Ga-Al-As laser irradiation eombined with MTX can effectively improve the clinical symptoms of RA patients. It may be related to the decrease of amount of PGE2 and count of CD4 + CD25 + regulatory T cell in PB and SF.