高级氧化蛋白产物在系统性红斑狼疮早发动脉粥样硬化中的临床意义

Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.     

高级氧化蛋白产物在系统性红斑狼疮早发动脉粥样硬化中的临床意义

Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.     

系统性红斑狼疮患者循环annexin Ⅱ水平的变化及临床意义

目的 探讨系统性红斑狼疮(SLE)患者血自细胞annexinⅡ的表达水平及其在SLE中的临床意义.方法 采用流式细胞术检测SLE患者35例,慢性肾炎患者10例,糖尿病肾病(DN)患者10例,健康对照组20名血白细胞annexinⅡ的表达水平,并分析其与临床指标间的相关性.采用t检验、方差分析及直线相关性分析进行统计学分析.结果 SLE、DN组患者血白细胞annexin Ⅱ水平[(7.1±2.9)%,(8.0+3.7)%]均显著低于健康对照组[(10.6±1.6)%](P<0.01,P<0.05);有活动性[(SLE疾病活动指数(SLEDAI)评分≥9分]的SLE患者annexinⅡ水平[(5.6±2.4)%]显著低于非活动性(SLEDAI评分<9分)的SLE患者[(7.8±2.8)%](P<0.05);SLE患者annexinⅡ水平与尿蛋白/肌酐呈负相关(r=-0.382,P<0.05),与血白蛋白呈正相关(r=0.439,P<0.01),与SLEDAI呈负相关(r=-0.417,P<0.05),与D-二聚体呈负相关(r=-0.336,P<0.05).结论 SLE患者血白细胞annexin Ⅱ表达水平降低,参与了SLE患者高凝和继发性纤溶亢进状态的发生发展的病理生理过程,可作为一种早期反映SLE血栓前状态的良好指标,对判断SLE的活动性及疗效有一定的帮助.

Abstract：

Objective To compare the level of Annexin Ⅱ in patients with systemic lupus erythematosus(SLE),diabetic nephropathy(DN),chronic glomerulonephritis and normal controls,and explorle the significance of the annexin Ⅱ in SLE.Methods Thirty-five cases of patients with SLE,ten cases of patients with DN,ten cases of patients with chronic glomerulonephritis were enrolled in this study,twenty cases of healthy controls were also enrolled.Circulating annexin Ⅱ in white blood cells was detected bv flow cytometry.Student's t test,variance analysis and Lineat correlation analysis were used for statistial analysis.Resuits Compared with healthy controls,the level of annexin Ⅱ in white blood cells in SLE patients (7.1±2.9)%and DN patients(8.0±3.7)%were significantly lower than that of the healthy controls(P<0.01,p<0.05).In the SLE group,the level of annexin Ⅱ of patients who had more active disease(SLEDAI≥9)decreased more thall those with less active disease(SLEDAI<9),(P<0.05).A positive correlation was found between annexin Ⅱ and serum albumin level(r:0.439,P<0.01),but negative correlation was found between annexin and urine protein/urine creatinine(r=-0.382,P<0.05),SLEDAI(r=-0.417,P<0.05),D-dimer(r=-0.336.p＜0.05) levels.Conclusion The level of annexin Ⅱ is decreased in patients with SLE,so it can renectthe abnormality of coagulation and fibrinolytic systems,and it may be used as a good indicator for prothrombotic status in SLE patients.It can be helpful to evaluatethe activity of the disease and the therapeutic efficacy.     

系统性红斑狼疮患者外周血单个核细胞CD226基因表达的研究

目的 研究CD226基因在系统性红斑狼疮(SLE)患者与健康人外周血单个核细胞(PBMCs)的表达及其与CD226-Gly307Ser多态性、疾病活动度的相关性.方法 应用实时荧光定量聚合酶链反应检测SLE患者组90例及健康对照组30名PBMCs CD226 mRNA表达水平,同时应用单因素方差分析CD226基因表达量在3种基因之间是否存在差异,并且探讨其与临床指标的相关性以及CD226-Gly307Ser多态性的3种基因型进行Pearson相关性分析.结果 SLE组与健康对照组相比,CD226基因表达水平明显降低(P＜0.01);SLE组3种基因型之间的CD226 mRNA表达量差异无统计学意义(6.8±1.1与26.5±6.7,P＞0.05);红细胞沉降率、尿蛋白定量(24 h)、抗核抗体滴度、SLE疾病活动指数(SLEDAI)及血清补体C3水平与CD226基因表达均没有相关性.结论 在中国湖北汉族人群中,CD226-Gly307Ser多态性与SLE发病相关;T危险等位基因未影响CD226的mRNA表达水平,CD226分子在自身免疫性疾病中作用需进一步探讨.

Abstract：

Objective To investigate the expression level of CD226 mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and explore the relation between the gene expression and disease activity, and the relation between the gene expression and Gly307Ser polymorphism of CD226 was also examined. Methods CD226 gene was measured with real-time polymerase chain reaction (qRT- PCR) in PBMCs. The expression levels of CD226 gene in PBMCs were compared between 90 SLE patients and 30 healthy individuals. One-way ANOVA and pearson correlation were used for statistical analysis. Results The expression level of CD226 in the PBMCs of SLE patients (6.8±1.1) was significantly decreased compared to healthy individuals (26.5±6.7) (P＜0.01), while there was no association between mRNA level and genotype (P＞0.05). No correlation between ESR, CRP, ANA, SLEDAI scores, C3 and the expression level of CD226 gene was discovered. Conclusion In Hubei Chinese Han population, CD226-Gly307Ser locus is associated with the development of SLE, while T allele does not impact the expression of CD226 gene, thus the role of CD226 gene in autoimmune diseases should be explored in the future.     

羟氯喹抑制紫外线诱导的系统性红斑狼疮CD4+T细胞白细胞介素-10和干扰素-γ的表达

目的 研究紫外线对系统性红斑狼疮(SLE)CD4+T细胞因子的影响和羟氯喹的抑制作用.方法 选择SLE 30例,健康对照10名.磁珠分选SLE患者和健康人的CD4+T细胞,紫外线311 nm窄谱中波紫外线暴露,加入羟氯喹共培养,酶联免疫吸附试验(ELISA)检测培养上清白细胞介素(IL)-10和干扰素-γ的表达水平.采用t检验进行统计学分析.结果 SLE患者CD4+T细胞IL-10表达高于健康对照[(27±4)和(18±3) pg/ml,P=0.011];经45、100 mJ/cm2紫外线暴露后,SLE活动患者CD4+T细胞IL-10表达升高[(27±4)和(77±42) pg/ml,(40±18)和(77±42) pg/ml,P=0.022,P=0.048],经100 mJ/cm2紫外线暴露后,活动患者CD4+T细胞IL-10表达高于稳定患者[(77±42)和(24±4)pg/ml,P=0.029];羟氯喹降低SLE活动患者CD4+T细胞IL-10和干扰素-γ表达[(2.6±4.0)和(17.9±2.3)pg/ml,P=0.018,P=-0.017)];羟氯喹降低经45,100 mJ/cm2紫外线暴露后SLE活动患者T细胞IL-10表达[(40±18)和(22±6)pg/ml,(77±42)和(21±5) pg/ml,P=0.037,P=0.04];羟氯喹降低经100 mJ/cm2紫外线暴露的SLE活动和稳定患者T细胞干扰素-γ表达[(18±3)和(13±14) pg/ml,(19±7)和(12±5) pg/ml,P=0.013,P=0.049].结论 紫外线加重SLE患者体内Th1/Th2细胞因子的比例失衡;羟氯喹抑制了紫外线诱发SLE患者干扰素-γ和IL-10的表达.

Abstract：

Objective To explore the role of hydroxychloroquine (HCQ) in ultraviolet B (UVB)- induced expression of interleukin (IL)-10 and interferon (IFN)-γ from CD4+T cells in patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 healthy controls were enrolled in the study. CD4+ T cells were isolated using magnetic beads from SLE patients and healthy controls. HCQ was added in culture media before and after irradiation with UVB 311 nm narrow band ultraviolet B (NB-UVB). The levels of IL-10 and IFN-γ in the supernatant were detected with enzyme-linked immunosorbent (ELISA). Comparisons between groups were performed by t-test. Results The level of IL-10 was higher in SLE patients [(27±4) pg/ml] than that in healthy controls [(18±3) pg/ml, P=0.011]. After exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages, the level of IL-10 was increased significantly in patients with active disease (P=0.022, P=0.048). After exposure of CD4+T cells to UVB in 100 mJ/cm2 dosages, the levels of IL-10 was higher in patients with active disease [(77±42) pg/ml] than patients with stable disease [(24± 4) pg/ml, P=0.029]. When CD4+ T cell were cultured with HCQ, IL-10 and IFN-γ levels in patients with active disease [(2.6±4.0), (17.5±2.3) pg/ml] were decreased significantly (P=0.018, P=0.017). HCQ reversed UVB-induced IL-10 expression in active SLE patients after exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages (P=0.037, P=0.04). HCQ also reversed UVB-induced IFN-7 expression in active SLE patients and stable SLE patients after exposure to CD4+T cells with UVB in 100 mJ/cm2 dosages (P=0.013, P= 0.049). Conclusion UVB can aggravate the imbalance of Th1 and Th2 cytokines. HCQ inhibits UVB-induced IL-10 and IFN-7 expression of CD4+T cells in patients with SLE, especially in patients with active disease.     

老年原发性非小细胞肺癌患者外周血T细胞膜型和可溶性CD28的表达

Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.     

老年原发性非小细胞肺癌患者外周血T细胞膜型和可溶性CD28的表达

Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.     

老年原发性非小细胞肺癌患者外周血T细胞膜型和可溶性CD28的表达

Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.     

干扰素诱导基因与系统性红斑狼疮的相关性研究

目的 探讨实时定量测定干扰素诱导基因[抗黏病毒1(MX1)基因,2′5′-寡腺苷酸合成酶1(OAS1),干扰素诱导蛋白44(IFI44)基因]表达水平与系统性红斑狼疮(SLE)临床表现及病情活动度的相关性.方法 收集100例SLE患者,40例非SLE其他自身免疫性疾病患者,40名健康对照的临床资料,取外周血抽提总RNA并反转录成cDNA,运用SYBR green dye Ⅰ实时定量聚合酶链反应(QT-PCR)在ABI PRISM 7000基因测序仪上检测患者和对照组的MXI、OAS1和IFI44定量表达水平(以△CT值表示)的差异,并与各临床指标及病情活动度进行相关性分析.采用方差分析和Spearman相关分析进行统计学处理.结果 ①SLE患者总体的MX1、OAS1、IFI44 mRNA △CT值(3.4±1.8,4.2±1.5,8.8±2.2)明显高于非SLE患者组(2.4±0.4,3.4±0.7,5.4±2.1)和健康对照组(2.3±1.2,2.6±0.7,5.2±2.0);②SLE患者组的OAS1、IFI44 mRNA △CT值在不同活动度的SLE组内[非活动组(3.8±1.4,3.2±1.8);轻度活动组(4.8±1.5,8.0±2.2);重度活动组(6.0±1.4,12.1±2.4)]差异具有统计学意义;③SLE患者组的OAS1、IFI44 mRNA△CT值与SLE疾病活动指数(SLEDAI)积分之间有相关性(r=0.338,0.380);④在有关节炎的SLE患者中,其MX1、OAS1、IFI44表达水平升高,与无关节炎患者比较差异具有统计学意义;⑤在狼疮肾炎组内,IFI44mRNA △CT值表达(2.2±1.1)与非狼疮肾炎组(3.2±2.1)比较差异有统计学意义;⑥SLE组内3个基因表达存在高相关性且具有统计学意义(P均＜0.05).结论 干扰素诱导基因MX1、OAS1、IFI44在SLE患者中均有表达上调现象,OAS1、IFI44 mRNA实时定量表达水平对SLE患者的病情活动度判断有意义,且三者在SLE的发生发展中亦存在密切联系.     

干扰素调节因子5在系统性红斑狼疮的表达及临床相关性研究

目的 比较干扰素调节因子5(IRF5)mRNA在系统性红斑狼疮(SLE)患者和健康对照组的表达水平,分析IRF5表达与SLE疾病活动性及自身抗体和临床症状的相关性.方法 Ficoll密度梯度离心法分离SLE患者及健康对照外周血单个核细胞(PBMC),Trizol法分离提取总RNA,反转录mRNA为eDNA;实时定量聚合酶链反应(PCR)法测定SLE患者和正常对照组IRF5表达量;并分析SLE患者IRF5表达量与疾病活动性及临床症状的相关性.结果 SLE组IRF5表达量(2.1+2.2)高于正常对照组(1.5±1.2),但差异无统计学意义(P=0.161);SLE患者IRF5表达量与其疾病活动指数(SLEDAI)显著相关(r=0.616,P<0.01);SLE患者中抗dsDNA抗体阳性组IRF5表达量(3.2±2.8)明显高于抗体阴性组(1.3±1.6).差异有统计学意义(P=0.018);SLE患者有发热、神经精神症状者IRF5表达量明显高于无此类临床症状者.结论 IRF5在SLE患者中表达偏高,且与SLEDAI显著相关,其可能通过调节下游基因的转录表达诱一导免疫失调,并由此参与SLE的发病过程.     

Th17细胞和调节性T细胞在系统性红斑狼疮患者中的研究

目的 研究系统性红斑狼疮(SLE)患者外周血Th17细胞、调节性T细胞及联系两者的细胞因子白细胞介素(IL)-6的变化及其与病情活动的相关性,探讨Th17细胞、调节性T细胞在SLE发病机制中的作用.方法 收集103例SLE患者及28名健康者,SLE活动性的判断采用SLE疾病活动指数(SLEDAI)评分方法,其中非活动组(SLEDAI≤9分)患者37例,活动组(SLEDAI＞9分)患者66例.用四色分选流式细胞仪检测外周血Th17细胞、调节性T细胞百分数,用流式细胞仪微球捕获芯片技术(CBA)检测血清IL-6浓度,并分析其与病情活动的相关性.统计学处理采用t检验、秩和检验和Spearman相关分析.结果 ①SLE患者组Th17细胞[(1.2±1.1)%]、IL-6[(35±92)pg/ml]显著高于健康对照组[(0.6±0.4)%、(6±3)pg/ml],而调节性T细胞[(1.6±1.2)%]显著低于健康对照组[(2.6±1.8)%].②SLE活动组Th17细胞[(1.6±1.7)%]显著高于SLE非活动组[(1.0±0.7)%]与健康对照组(P均＜0.05),SLE活动组调节性T细胞[(1.5±1.3)%]显著低于SLE非活动组[(2.1±2.0)%]与健康对照组(P均＜0.05),SLE活动组Th17/调节性T细胞(0.68±0.34)显著高于SLE非活动组(0.24±0.20)与健康对照组(0.13±0.09,P均＜0.05),SLE活动组IL-6[(41±22) pg/ml]显著高于SLE非活动组[(32±28) pg/ml]与健康对照组(P均＜0.05);SLE非活动组与健康对照组之间Th17细胞、调节性T细胞、Th17/调节性T细胞、IL-6差异均无统计学意义(P＞0.05).③Th17细胞百分数与SLEDAI呈正相关,调节性T细胞百分数与SLEDAI呈负相关,Th17/调节性T细胞与SLEDAI呈正相关,IL-6浓度与SLEDAI评分之间呈正相关,Th17细胞与调节性T细胞呈负相关.结论 SLE患者外周血Th17细胞、调节性T细胞及IL-6浓度较健康对照组显著增高,并与疾病活动性密切相关,说明Th17细胞、调节性T细胞可能在SLE的发生、发展中起重要作用.

Abstract：

Objective To study the changes of Th17 cell, regulatory T cell (Treg) and interleukin (IL)-6 in the peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with disease activity. Methods Percentage of Th17 and Treg in the peripheral blood of 103 patients with SLE and 28 healthy volunteers were detected by flow cytometry. The concentration of IL-6 in SLE patients and healthy volunteers was detected by cytometric bead array (CBA). The disease activity of SLE was measured by SLEDAI. SLE patients were divided into two groups: stable SLE (SLEDAI≤ 9, n=37) and active SLE (SLEDAI＞9, n= 66). The change of Th17, Treg, IL -6 and their relationship with disease activity were analyzed. Nonparamentric tests, t -test and spearman correlation were used for statistical analysis. Results The percentage of Th17 cells and the concentration of IL-6 in the peripheral blood in patients with SLE was higher than that in normal controls [respectively for (1.2±1.1)%, (35±92) pg/ml and (0.6±0.4)%, (6±3) pg/ml, P＜0.05]. However, the percentage of Treg in patients with SLE was lower than that in normal controls [respectively for (1.6±1.2)%,(2.6±1.8)%, P＜0.05]. The percentage of Th17, Th17/Treg IL-6 level in active SLE patients was higher than those in inactive SLE and those in normal controls (P＜0.05). However, the percentage of Treg in active SLE was lower than that in stable SLE patients and that in normal controls (P＜ 0.05). The percentage of Th17, Th17/Treg and concentration of IL-6 was positively correlated to disease activity(P＜0.05). But the percentage of Treg had negative correlation with the percentage of Th17 and disease activity (P＜0.05). Conclusion Th17, Treg and serum IL-6 in SLE patients are abnormal and they maybe contribute to the pathogenesis of SLE.     

Toll样受体9与系统性红斑狼疮患者的临床特点相关性

目的 探讨Toll样受体9（toll-like receptor 9,TLR9）在系统性红斑狼疮（systemic lupus erythematosus,SLE）患者中的表达情况及其与临床的关系.方法 采用细胞内流式细胞术对48例SLE组、36例疾病对照组（包括类风湿关节炎、干燥综合征、系统性硬化症、肌炎/皮肌炎）和25名健康对照组患者外周血B淋巴细胞中TLR9表达情况进行比较,并记录SLE患者临床表现和实验室检查,对TLR9的表达与临床特点的相关性进行分析.结果 SLE活动期组外周血CD19＋B细胞TLR9表达率（54.18±17.97）高于SLE非活动期（32.97±9.85）、疾病对照组（30.88±9.12）及健康对照组（29.75±7.49）（均P〈0.01）.且其表达率与SLE疾病活动指数评分（r=0.763）、抗双链DNA抗体滴度（r=0.781）呈正相关（均P〈0.01）.应用中小剂量糖皮质激素对SLE活动期患者TLR9在CD19＋B细胞的表达无显著差异（P〉0.05）.结论 TLR9参与了SLE的发病,可以作为判断疾病活动性及临床预后的指标.     

Increased apoptotic peripheral blood neutrophils in systemic lupus erythematosus: relations with disease activity, antibodies to double stranded DNA, and neutropenia

OBJECTIVE: To quantify the percentage of apoptotic peripheral blood neutrophils in systemic lupus erythematosus (SLE) and to determine the relations with disease activity and neutropenia. METHODS: Neutrophil apoptosis in SLE patients (n =50) was assessed by flow cytometry using annexin V binding and fluorescent labelled anti-fas. Rheumatoid arthritis (RA, n =20) and inflammatory bowel disease patients (IBD, n =20) were studied as disease controls. RESULTS: The percentage of apoptotic neutrophils, determined by annexin V binding, was increased in peripheral blood of SLE patients (median = 3.25%) compared with normal healthy donors (n =20, median = 1.20%) and disease controls (RA: median = 1.15%) (IBD: median = 1.15%). SLE neutrophil apoptosis correlated positively with lupus disease activity measured by SLAM score. SLE patients with increased antibodies to dsDNA (>10 mg/ml) had increased apoptotic neutrophils. Eight of 14 neutropenic SLE patients had increased apoptotic neutrophils. Increased neutrophil fas expression compared with normal controls was observed in SLE, RA, and IBD. CONCLUSION: Neutrophil fas expression is increased non-specifically in inflammatory disease. Increased circulating apoptotic neutrophils in SLE correlate positively with disease activity (SLAM) and may contribute to autoantigen excess including dsDNA.     

系统性红斑狼疮患者外周血CD4+CD25highFoxp3+调节性T细胞数量和Foxp3mRNA的表达水平与疾病活动性的相关性

目的 研究系统性红斑狼疮(SEE)患者CD4+CD25highFoxp3+调节性T细胞的数量及其功能基因Foxp3 mRNA的表达水平与SLE疾病活动性和肾脏损伤的相关性.方法 采用四色流式细胞术以Foxp3-异硫氰酸荧光素(FITC )/CD25-藻红蛋白/CD4-多甲藻叶绿素蛋白(PerCP)/CD3-藻蓝蛋白7抗体组合检测40名健康对照者及42例SLE患者外周血CD4+CD25highFoxp3+调节性T细胞的数量,实时荧光定量聚合酶链反应(PCR)检测特异性转录因子Foxp3 mRNA的表达水平,并分析其与SLE患者疾病活动指数(SLEDAI)、补体C3及血清抗双链DNA(dsDNA)抗体的关系.统计学方法采用t检验和Spearman相关分析.结果 活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量显著低于健康对照组[(4±3)％与(7±4)％,P＜0.05],稳定期与健康对照组差异无统计学意义(P＞0.05);活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于稳定期患者[(4±3)％,(9±6)％与(5±4)％,(10±6)％,P均＜0.05];活动期SLE患者外周血Foxp3 mRNA的表达水平明显低于稳定期和对照组(P＜0.01,P＜0.05);SLE患者并发肾病组外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于SLE非肾病组(P＜0.05).相关分析显示,SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与SLEDAI呈负相关(r=-0.5782,P＜0.05);CD4+CD25highFoxp3+调节性T细胞/CD4+比值与SLEDAI呈负相关(r=-0.4913,P＜0.05),与补体C3呈正相关(r=0.3687,P＜0.05);SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与Foxp3 mRNA的表达水平呈正相关(r=0.6142,P＜0.0l).结论 SLE患者外周血CD4+CD25highFoxp3+调节性T细胞和Foxp3 mRNA的变化可能是导致SLE疾病发生和发展的关键因素之一,与疾病的活动性有密切关系.     

炎症性肠病患者CD40-CD154共刺激途径的激活及其与内镜下疾病活动程度的相关性研究

目的探讨CD40-CD154共刺激途径在炎症性肠病患者外周循环和肠黏膜中的表达,比较炎症性肠病患者CD40-CD154表达和正常对照者的差异,分析CD40-CD154表达与内镜下疾病活动性的相关性。方法研究对象为克罗恩病患者62例、溃疡性结肠炎患者64例和正常对照者56例。对炎症性肠病患者和正常对照者,分别应用酶联免疫吸附试验（ELISA）、SYBR—green real time PCR方法、免疫组化法,分析血浆中、外周血单个核细胞中、肠黏膜组织中CD40-CD154的表达情况。结果克罗恩病、溃疡性结肠炎患者血浆、外周血单个核细胞及肠黏膜组织中,CD40和CD154的表达均显著高于正常对照者（P均〈0．05）,但外周循环和肠黏膜中CD40及CD154的表达和内镜下疾病活动性无关（P均〉0．05）。结论炎症性肠病患者血浆、外周血单个核细胞和肠黏膜中均存在CD40-CD154途径的激活,但CD40-CD154的高表达不能反映内镜下疾病活动程度。     

趋化因子CXCL13在系统性红斑狼疮患者的血浆表达水平及临床意义

目的 检测系统性红斑狼疮(SLE)患者血浆趋化因子CXCL13表达,分析CXCL13表达与SLE疾病活动性及脏器受累的关系,探讨CXCL13在SLE发病机制中的可能作用.方法 酶联免疫吸附法(ELISA)检测60例SLE患者血浆CXCL13水平,并与21名健康体检者对照.组间计量资料比较采用Mann-Whitney检验,组间计量资料相关性分析采用Pearson相关性分析.结果 血浆趋化因子CXCLl3在SLE患者组表达水平明显高于健康对照组[(501±446)和(102±121) pg/ml,P＜0.01].有肾脏及中枢神经系统受累的SLE患者较无相应器官受累的SLE患者有着较高水平的血浆CXCL13表达(P=0.038,0.026).另外,分析发现血浆CXCL13表达水平与SLE疾病活动指数呈正相关(r=0.585,P=0.001).结论 趋化因子CXCL13在SLE发病机制中具有重要作用,可能参与了SLE肾脏及中枢神经系统受累过程;CXCL13可作为判断SLE病情活动性的指标.     

信号转导与转录激活因子2基因在外周血中的高表达与系统性红斑狼疮的病情活动相关

目的 探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中信号转导与转录激活因子2(STAT2)实时定量表达与SLE疾病特异性和病情活动度的相关性.方法 收集144例SLE患者、27例非SLE患者与58名健康对照者的临床资料,取外周血抽提总RNA并逆转录成cDNA,运用SYBR green dye Ⅰ实时定量聚合酶链反应(real-time MR)法检测患者组和对照组的STAT2 mRNA表达水平的差异,并与病情活动度进行分组比较,分析其意义.结果 SLE患者组的STAT2定量表达(5.2±1.7)高于非SLE患者组和健康对照组(4.3±1.1,4.5±1.2,P均<0.01);SLE活动组的STAT2表达(5.2±1.5)高于非活动组(4.8±2.9,P<0.01);SLE患者组的STAT2表达与SLE疾病活动指数(SLEDAI)-2000和尿蛋白值之间呈正相关(r=0.317,0.309,P均<0.01),与血清补体C3水平呈负相关(r=-0.449,P<0.01).结论 转录因子STAT2在外周血细胞中的异常表达与SLE的发病机制有关,STAT2 mRNA定量表达水平的升高与SLE患者病情活动相关.     

血浆可溶型人类白细胞抗原-G在系统性红斑狼疮患者中的表达分析

目的 检测系统性红斑狼疮(SLE)患者血浆可溶性人类白细胞抗原-G(sHLA-G)水平,并分析其与SLE脏器受累及疾病活动性的关系,探讨sHLA-G在SLE发病机制中的可能作用.方法 酶联免疫吸附法(ELISA)检测96例SLE患者血浆sHLA-G水平,并与74名健康体检者对照.采用t检验,直线相关回归方法 和X2检验,P<0.05为差异有统计学意义.结果血浆sHLA-G水平在SLE患者为(230±192)U/ml;显著高于健康体检者的(118±38)U/ml(t=5.07,P=0.0001);血浆sHLA-G水平与SLE疾病活动性指数(SLEDIA)无明显相关性(r=0.157,P=0.141);但血浆sHLA-G水平增高的SLE患者较血浆sHLA-G水平正常患者SLEDAI高(11±5与8±5,P=0.027),易出现中枢神经系统受累(24.2%与4.8%,P=0.007).结论 血浆sHLA-G水平增高SLE患者病情较重、中枢神经系统受累较多,提示sHLA-G在SLE病理过程中可能发挥重要作用.