TNF-α在强直性脊柱炎患者血清中的表达

肿瘤坏死因子一仅（TNF-α）是一种具有杀伤肿瘤细胞功能的促炎细胞因子,主要由活化的单核巨噬细胞产生,具有多种生物学功能,在炎症反应、免疫调节中起重要作用。在免疫病理发病机制的研究中发现,强直性脊柱炎（AS）的发病与促炎细胞因子表达的上调有关,其中较为重要并为多数学者所公认的细胞因子是TNF-α。     

湖南地区强直性脊柱炎患者TNFRⅡ基因多态性研究

目的 通过对湖南地区强直性脊柱炎(AS)汉族患者及健康人群DNA中肿瘤坏死因子受体(TNFR)Ⅱnt587位点的多态性分析,研究TNFRⅡnt587位点基因多态性与强直性脊柱炎发病的相关性.方法 采用聚合酶链反应-限制性片段长度多态性法(PCR-RFLP)对100例AS患者和100名健康对照TNFRⅡnt587位点进行基因分型检测.数据经SPSS处理,分析两组中基因型、等位基因频率差异.结果 100例患者中TNFRⅡn687位点T/T型43例(43.0％),T/G型32例(32.0％),G/G型25例(25.0％);100例健康对照中TNFRⅡnt587位点T/T型56例(56.0％),T/G型34例(34.0％),G/G型10例(10.0％),AS患者中TNFRⅡnt587 G频率(41.0％)较正常对照(27.0％)明显增高(x2=8.734,P=0.003).与对照组相比,AS患者TNFRⅡnt587位点基因型为G/G时比数比(OR=3.256)明显高于T/G型(OR=1.226)和T/T型(OR=1).结论 湖南地区汉族人群中TNFRⅡnt587位点多态性与AS发病具有相关性,且G/G基因型在AS发病中具有高风险性.     

肿瘤坏死因子-α 5 旁侧基因多态性与强直性脊柱炎相关性

目的探讨肿瘤坏死因子α基因多态性与强直性脊柱炎(AS)发生的易感关系。方法采用序列特异引物PCR方法(SSP-PCR)检测了136例AS患者和127例正常人的TNF-α5旁侧-857C/T,-863C/A和-1031T/C三个基因多态性位点,并比较了部分不同地区人群之间的基因型与等位基因频率。结果AS患者TNF-α-857基因型(CC,TC和TT)分布频率分别是64.0%,35.3%和0.7%;TNF-α-863基因型(CC,CA和AA)分布频率分别是68.4%,30.0%和1.5%;TNF-α-1031基因型(TT,TC和CC)分布频率分别是66.2%,32.4%和1.5%;TNF-α-857等位基因频率C和T分别是81.6%和18.8%;TNF-α-863等位基因频率C和A分别是83.5%和16.5%;TNF-α-1031等位基因频率T和C分别是:82.4%和17.6%;正常对照者TNF-α-857基因型(CC,TC和TT)分布频率分别是70.9%,27.6%和1.6%;TNF-α-863基因型(CC,CA和AA)分布频率分别是68.5%,29.9%和1.6%;TNF-α-1031基因型(TT,TC和CC)分布频率分别是69.3%,29.1%和1.6%;TNF-α-857等位基因频率C和T分别是84.6%和15.4%;TNF-α-863等位基因频率C和A分别是83.5%和16.5%;TNF-α-1031等位基因频率T和C分别是:83.9%和16.1%;AS患者和正常对照者之间的基因型分布和等位基因频率比较差异无显著性,(P>0.05);本文中国汉族人TNF-α-857(C/T),-863(C/A),-1031(T/C)位点基因多态性分布同亚洲的日本、韩国人群基本一致,差异无显著性;-863(C/A),-1031(T/C)位点基因多态性分布同欧洲的英国、荷兰、瑞典人群比较差异也无显著性,但欧洲人群的TNF-α-857T的等位基因频率明显高于亚洲人群,差异有显著性(P<0.05)。结论TNF-α-857,-863和-1031三个基因多态性位点可能不是中国人患AS的主要易感位点基因。     

CD40-CD40L在强直性脊柱炎患者外周血淋巴细胞上的表达

目的探讨共刺激分子CD40CD40L在强直性脊柱炎(AS)患者的外周血淋巴细胞亚群上的异常表达与免疫功能紊乱的关系。方法用流式细胞仪采用直接免疫荧光法测定30例AS患者和20例健康对照人外周血淋巴细胞表面标志CD3、CD4、CD8、CD19的表达情况,CD40L在CD4 T和CD8 T细胞上的表达及CD40在CD19 B细胞上的表达。用速率散射比浊法测定血清中免疫球蛋白IgG,IgA和IgM的水平。结果①AS患者CD3 、CD3 CD4 、CD19 细胞较正常对照组显著增高(P<0.05),CD3 CD8 细胞较正常对照组显著降低(P<0.05);②AS患者CD4 T细胞和CD8 T细胞上的CD40L、CD19 B细胞上CD40的表达都较对照组显著增高(P<0.05);③AS患者血清中2种免疫球蛋白IgG、IgA的水平均较对照组显著增高(P<0.05)。结论CD40CD40L途径在AS免疫功能紊乱中起了重要作用。     

IL-13 induces expression of CD36 in human monocytes through PPARgamma activation

The class B scavenger receptor CD36 is a component of the pattern recognition receptors on monocytes that recognizes a variety of molecules. CD36 expression in monocytes depends on exposure to soluble mediators. We demonstrate here that CD36 expression is induced in human monocytes following exposure to IL-13, a Th2 cytokine, via the peroxisome proliferator-activated receptor (PPAR)gamma pathway. Induction of CD36 protein was paralleled by an increase in CD36 mRNA. The PPARgamma pathway was demonstrated using transfection of a PPARgamma expression plasmid into the murine macrophage cell line RAW264.7, expressing very low levels of PPARgamma, and in peritoneal macrophages from PPARgamma-conditional null mice. We also show that CD36 induction by IL-13 via PPARgamma is dependent on phospholipase A2 activation and that IL-13 induces the production of endogenous 15-deoxy-Delta12,14-prostaglandin J2, an endogenous PPARgamma ligand, and its nuclear localization in human monocytes. Finally, we demonstrate that CD36 and PPARgamma are involved in IL-13-mediated phagocytosis of Plasmodium falciparum-parasitized erythrocytes. These results reveal a novel role for PPARgamma in the alternative activation of monocytes by IL-13, suggesting that endogenous PPARgamma ligands, produced by phospholipase A2 activation, could contribute to the biochemical and cellular functions of CD36.     

Evaluation of serum paraoxonase and arylesterase activities in ankylosing spondylitis patients

OBJECTIVES

The aim of this study was to investigate the activities of serum paraoxonase and arylesterase in patients with ankylosing spondylitis with respect to those of healthy controls, to assess whether these enzyme levels are related to disease activity and functional capacity.

METHODS

The study included 32 patients with ankylosing spondylitis whose diagnoses were made according to the modified New York criteria as well as 25 healthy controls matched for age and sex. The Bath Ankylosing Spondylitis Disease Activity Index and the Bath Ankylosing Spondylitis Functional Index were applied to the ankylosing spondylitis patients. As laboratory parameters, the erythrocyte sedimentation rate and serum C-reactive protein level were measured in patients and control subjects. Paraoxonase and arylesterase enzyme activities were measured using appropriate methods.

RESULTS

No statistically significant differences (p>0.05) were found between the ankylosing spondylitis patients and controls in terms of serum paraoxonase or arylesterase levels. Furthermore, there was no correlation between clinical and laboratory parameters in patients with ankylosing spondylitis.

CONCLUSION

Serum paraoxonase and arylesterase levels in ankylosing spondylitis patients may not differ from those of healthy controls, and there is no significant correlation between antioxidant parameters and the Bath Ankylosing Spondylitis Disease Activity Index or Bath Ankylosing Spondylitis Functional Index scores in ankylosing spondylitis patients. Further research is needed to provide deeper understanding of this disease.     

ciglitazone对巨噬细胞CD36表达及胆固醇流入的影响

目的:观察过氧化物酶体增殖物激活型受体γ(PPARγ)的激动剂ciglitazone对THP-1巨噬细胞CD36表达及细胞胆固醇流入的影响。方法:以THP-1巨噬细胞为研究对象,用50 mg/L氧化低密度指蛋白(oxLDL)分别与不同浓度的PPARγ激动剂ciglitazone(0、5、10、15μmol/L)共同孵育THP-1巨噬细胞24 h后,采用液体闪烁计数法检测[3H]胆固醇流入,采用逆转录-聚合酶链式反应和Western blot分别检测CD36的表达。结果:oxLDL及不同浓度的ciglitazone与THP-1巨噬细胞共孵育后,与对照组比较(20.3%),加ciglitazone的处理组胆固醇流入随着浓度的增加而依次增加,分别为28.6%、37.2%、44.3%、48.7%,细胞CD36的表达上调。结论:ciglitazone可上调THP-1巨噬细胞CD36的表达,并增加细胞胆固醇流入。     

Serum from patients with ankylosing spondylitis can increase PPARD,fra-1, MMP7, OPG and RANKL expression in MG63 cells

OBJECTIVES:To explore the effects of serum from patients with ankylosing spondylitis on the canonical Wnt/β-catenin pathway and to assess whether the serum has an osteogenic effect in MG63 cells.METHODS:MG63 cells were cultured with serum from 45 ankylosing spondylitis patients, 30 healthy controls, or 45 rheumatoid arthritis patients. The relative PPARD, fra-1, MMP7, OPG and RANKL mRNA levels were measured using quantitative real-time polymerase chain reaction. Associations between gene expression and patient demographics and clinical assessments were then analyzed.RESULTS:MG63 cells treated with serum from ankylosing spondylitis patients had higher PPARD, fra-1, MMP7 and OPG gene expression than did cells treated with serum from controls or rheumatoid arthritis patients (all p<0.05). RANKL expression was higher in MG63 cells treated with serum from patients with ankylosing spondylitis or rheumatoid arthritis than in those treated with serum from controls (both p<0.05). The OPG/RANKL ratio was also higher in MG63 cells treated with serum from ankylosing spondylitis patients than in those treated with serum from controls (p<0.05). No associations were found between the expression of the five genes and the patient demographics and clinical assessments (all p>0.05).CONCLUSIONS: Serum from ankylosing spondylitis patients increases PPARD, fra-1, MMP7, OPG and RANKL expression and the OPG/RANKL ratio in MG63 cells; these effects may be due to the stimulatory effect of the serum on the Wnt pathway.     

Safety and clinical responses in ankylosing spondylitis after three months of etanercept therapy

We aimed to evaluate the safety and clinical responses in Korean ankylosing spondylitis (AS) patients after three months of etanercept therapy. AS patients satisfying the Modified New York Criteria were enrolled. They were assessed for safety and clinical responses at enrollment and after three months of etanercept therapy. A total of 124 patients completed the study. After three months, the rate of ASsessment in AS International Working Group 20% improvement (ASAS 20) response was 79.8%. The rates of ASAS 40 and ASAS 5/6 responses were 58.5 and 62.8%, respectively. Significant improvement of Korean version of Bath AS Disease Activity Index (KBASDAI) (p<0.0001), Bath AS Functional Activity Index (BASFI) (p<0.0001), and Bath AS Metrology Index (BASMI) (p=0.0009) were achieved after three months. Quality of life was also significantly improved after three months, as demonstrated by scores for SF-36 (p<0.0001) and EQ-5D (p<0.0001). Erythrocyte sedimentation rate and C-reactive protein were significantly decreased (p<0.0001, p<0.0001, respectively). None of the patients developed tuberculosis and there were no serious adverse event. AS patients with inadequate response to conventional therapy showed significant clinical improvement without serious adverse events after three months of etanercept therapy.     

强直性脊柱炎患者外周血CD4+调节性T细胞的变化及意义

目的 探讨强直性脊柱炎(ankylosing spondylitis,AS)患者外周血中CD4+调节性T细胞(regulatory T cells,Treg)的表达、功能及意义.方法 采用流式细胞术检测78例AS患者和50例健康志愿者外周血中CD4+CD25+CD127lo/- Treg、细胞毒性T细胞(cytotoxic T lymphocytes,CTL)和NK细胞,采用ELISA法检测血清中β型转化生长因子(transforming growth factor-β,TGF-β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达水平.从患者外周血单个核细胞中磁珠分选出CD4+CD25+ Treg细胞,混合淋巴细胞培养(mixed lymphocyte culture, MLC)分析其免疫抑制功能.结果 AS活动期患者外周血中CD4+CD25+CD127lo/-Treg占CD4+ T淋巴细胞的百分比为(4.36±1.21)%,MLC中CD4+CD25+ Treg抑制同种异体T淋巴细胞增殖功能低下,与其Treg分泌TGF-β减少相关,CD4+ Treg含量及功能均低于健康志愿者(P<0.05).AS患者外周血中CD4+CD25+CD127lo/- Treg水平与TGF-β呈正相关,与TNF-α呈负相关.结论 AS患者外周血中Treg表达水平低,且存在功能缺陷,导致体内诱导免疫耐受机能不足,可能参与AS免疫发病.     

Widespread scleredema accompanied with a monoclonal gammopathy in a patient with advanced ankylosing spondylitis

Scleredema is a rare cutaneous mucinosis characterized by chronic diffuse induration of the skin, and it is occasionally associated with a monoclonal gammopathy (MG). Ankylosing spondylitis (AS) is noted to be another, chronic systemic inflammatory disorder of the axial skeleton that may accompany the MG. However, patients with scleredema and AS accompanied with a MG have not been reported in the literature. We here report a 40-yr-old man with scleredema and advanced AS accompanied with a MG of IgA-kappa protein. Widespread, long-standing scleredema has been developed over 10 yrs after the initial manifestation of AS. It is uncertain whether the coexistence of scleredema and AS is more than coincidental.     

M2 polarization of monocytes in ankylosing spondylitis and relationship with inflammation and structural damage

The aim of this study was to evaluate the polarization of peripheral blood monocytes in the patients with ankylosing spondylitis (AS) and to determine the correlations between monocyte polarization and inflammation and structural damage. A total of 120 AS patients, 50 rheumatoid arthritis (RA) patients and 100 healthy controls were enrolled in the study. M1 (CD68+CD192+) and M2 (CX3CR1+CD163+) monocytes were characterized by flow cytometry. Demographic, clinical, radiographic and laboratory data were collected and analyzed. A large increase in M2 (CX3CR1+CD163+) monocytes was observed in AS, and M2/M1 ratio was 7.18 ± 6.12, 2.54 ± 3.14 and 35.61 ± 20.04 in control, RA and AS, respectively. The M2/M1 ratio correlated with modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) (r = 0.565; p < 0.001), ESR (r = ?0.321; p < 0.001, CRP (r = ?0.265; p < 0.001) and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) (r = ?0.201; p = 0.028). Anti‐TNF‐α therapy induced a significant reduction in the percentage of M1 monocyte, ESR, CRP and BASDAI in AS patients. The present results demonstrated that M2 type polarized monocytes are predominant in the peripheral blood in AS and the M2/M1 ratio is correlated with structural damage (mSASSS), inflammatory biomarkers (ESR and CRP) and BASDAI.     

过氧化物酶体增殖体活化受体γ、肝脏X受体α、视黄酸X受体α活化对THP-1细胞ATP结合盒转运蛋白和CD36mRNA表达的影响

目的:探讨核受体过氧化物酶体增殖体活化受体γ(PPARγ)、肝脏X受体α(LXRα)、视黄酸X受体α(RXRα)活化对THP1单核细胞ATP结合盒转运蛋白(ABC1)和CD36mRNA表达影响的差异及其意义。方法:体外培养的THP-1单核细胞,经氟波酯(PMA)诱导48h,使形成巨噬细胞,再分别用核受体PPARγ、LXRα、RXRα的相应配体曲格列酮、22(R)-羟化胆固醇[22(R)-HC],9-顺式-视黄酸(9-CRA)作用24h,收集细胞,提取总RNA,以β-actin作内参,用RT-PCR法检测ABC1和CD36mRNA表达的变化。结果:核受体PPARγ、LXRα、RXRα活化剂均可致ABC1mRNA表达增高,但对CD36mRNA的表达存在明显差异。PPARγ活化时CD36mRNA表达增高,LXRα活化时CD36mRNA表达并无明显增高,而RXRα活化时CD36mRNA表达降低。结论:核受体PPARγ、LXRα、RXRα活化对ABC1和CD36mRNA表达的影响存在差异,提示LXRα、RXRα可能是促进胆固醇逆向转运更理想的干预靶点。     

CD36与动脉粥样硬化的相关性研究

目的 探讨清道夫受体CD36在动脉粥样硬化患者血清中的变化及CD36在人主动脉粥样硬化组织中的表达情况,并研究两者的相关性及其临床意义.方法 选择心血管内科冠心病患者(CHD) 100例作为冠心病组,其中包括稳定型心绞痛(SAP)31例,不稳定型心绞痛(UAP) 29例,急性心肌梗死(AMI)40例.同时选择健康体检者50例作为对照组.ELISA法对两组血清进行CD36的浓度检测,检测其甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、hs-CRP、同型半胱氨酸(Hcy)含量,并对结果数据进行统计学分析.选择经冠脉造影诊断为冠心病的心脏手术患者的冠状动脉组织,同时选择因意外伤害需要心脏手术患者的冠状动脉组织作为对照组,免疫组化法检测两组组织中的CD36的表达.结果 ELISE法检测健康对照组CD36的含量为3.81 ±0.95pg/mL,而冠心病患者血清中CD36含量为25.17±3.76pg/mL,冠心病患者血清中CD36含量较健康对照组明显升高,差异有统计学意义(P＜0.05).其中稳定型心绞痛(SAP)CD36含量为18.68 ±2.17 pg/mL,不稳定型心绞痛(UAP)24.19±0.82 pg/mL,急性心肌梗死(AMI)27.17±5.83 pg/mL,不稳定型心绞痛和急性心肌梗死组CD36水平较稳定型心绞痛的明显升高(P＜0.01).动脉粥样硬化患者组CD36表达明显高于健康对照组,差异有统计学意义(x2 =23.018,P＜0.05).结论 清道夫受体CD36在动脉粥样硬化患者血清中浓度明显升高,且在动脉粥样硬化患者组织中高表达,临床上动脉粥样硬化患者检测CD36具有重要临床意义.     

强直性脊柱炎患者血清白细胞介素18检测的意义

目的探讨白细胞介素18(IL-18)在强直性脊柱炎(AS)发病中的作用及其血清水平检测的临床意义。方法采用酶联免疫吸附试验法(ELISA)检测53例活动期AS患者与40例健康对照组血清IL-18及肿瘤坏死因子α(TNF-α)的水平,并动态观察治疗前后IL-18及TNF-α的变化及其与病情活动指标之间的关系。结果AS患者治疗前血清IL-18水平明显高于健康对照组[(415.7±30.4)ng/L比(45.7±10.4)ng/L,P<0.05)],并与TNF-α、Bath强直性脊柱炎病情活动性指数(BASDAI)、晨僵时间、Bath强直性脊柱炎功能指数(BASFI)和Bath强直性脊柱炎脊柱活动测量指数(BASMI)呈正相关(r分别为0.52、0.63、0.48、0.58和0.48,P均<0.01)。治疗12周、24周后AS患者血清IL-18水平分别为(323.6±85.8)和(220.1±15.1)ng/L,均较治疗前明显下降,且治疗24周后下降更明显。结论IL-18可能参与AS发病机制,血清IL-18的检测可作为反映AS病情活动的指标之一。     

强直性脊柱炎患者血清白细胞介素18检测的意义

目的 探讨白细胞介素18(IL-18)在强直性脊柱炎(AS)发病中的作用及其血清水平检测的临床意义.方法 采用酶联免疫吸附试验法(ELISA)检测53例活动期AS患者与40例健康对照组血清IL-18及肿瘤坏死因子α(TNF-α)的水平,并动态观察治疗前后IL-18及TNF-α的变化及其与病情活动指标之间的关系.结果 AS患者治疗前血清IL-18水平明显高于健康对照组[(415.7±30.4)ng/L比(45.7±10.4)ng/L,P＜0.05)],并与TNF-α、Bath强直性脊柱炎病情活动性指数(BASDAI)、晨僵时间、Bath强直性脊柱炎功能指数(BASFI)和Bath强直性脊柱炎脊柱活动测量指数(BASMI)呈正相关(r分别为0.52、0.63、0.48、0.58和0.48,P均＜0.01).治疗12周、24周后AS患者血清IL-18水平分别为(323.6±85.8)和(220.1±15.1)ng/L,均较治疗前明显下降,且治疗24周后下降更明显.结论 IL-18可能参与AS发病机制,血清IL-18的检测可作为反映AS病情活动的指标之一.     

白介素6和白介素8对单核细胞组织因子表达的影响

白介素 6(ＩＬ 6)是一种多功能的细胞因子 ,除涉及感染、炎症等急性期反应外 ,对Ｔ、Ｂ细胞的发育、骨髓造血细胞增殖均有很重要的作用[1] 。自多发性骨髓瘤患者血浆ＩＬ 6水平增高被发现后 ,又发现ＩＬ 6血浆水平增高还见于弥散性血管内凝血 (ＤＩＣ) ,并且与ＤＩＣ的严重程度     

IL-23与IL-17在强直性脊柱炎患者中表达的初步研究

目的:通过研究IL-23与IL-17在强直性脊柱炎(AS)患者中的表达情况,为进一步阐明As发病机制和寻找新的治疗靶点提供理论依据.方法:AS患者与健康对照血清及培养的外周血单个核细胞(PBMCs)上清中IL-23与IL-17水平应用ELISA方法检测;应用RT-PCR方法检测PBMCs中IL-23p19 mRNA的表达.结果:39例活动期AS患者血清IL-23与IL-17水平分别为(1 159.71±139.45)pg/ml和(172.21±73.81)pg/ml,均较健康对照明显升高(P<0.001);AS患者培养的PBMCs上清IL-23与IL-17水平分别为(108.63±34.53)pg/ml和(134.59±38.32)pg/ml,明显高于健康对照(P<0.001),IL-23p19 mRNA表达明显高于健康对照,平均光密度分析差异有极显著统计学意义(P<0.001);IL-23可促进健康对照和AS患者的PBMCs IL-17的分泌,此作用在AS患者更显著.结论:IL-23与IL-17可能在AS的发病中发挥作用,IL-23可能通过诱导IL-17的产生而使后者在AS的发病中发挥作用.     

单核细胞与内皮细胞的相互作用对单核细胞CD36表达的影响

摘要目的:研究单核细胞、内皮细胞的相互作用对单核细胞清道夫受体CD36抗原表达的影响并在此基础上观察细胞因子在其中的介导作用。方法:使用单核细胞、内皮细胞单独培养及混合培养的方法形成不同的培养条件,通过ABC-ELISA法检测培养液中巨噬细胞集落刺激因子(M-CSF)的含量,使用流式细胞术检测单核细胞表面CD36抗原的表达程度。结果:单核细胞单独培养条件下CD36的表达量较低,在和内皮细胞混合培养后CD36表达量显著升高(P＜0.05),M-CSF和血小板源性生长因子(PDGF-BB)在单核细胞CD36表达增高的调节中起着一定的介导作用,但不占主要地位。结论:单核细胞与内皮细胞的相互作用可通过多种环节上调单核细胞CD36的表达,从而参与动脉粥样硬化的发展。