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.

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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.     

三七皂苷对系统性红斑狼疮患者外周血淋巴细胞P-糖蛋白及激素效应的影响

目的 探讨系统性红斑狼疮(SLE)患者糖皮质激素耐药机制以及三七皂苷逆转耐药的作用.方法 直线相关分析SLE患者外周血淋巴细胞P-糖蛋白与SLE疾病活动指数(SLEDAI)、临床疗效的相关性.用三七皂苷干预患者外周血淋巴细胞,维拉帕米为对照,流式细胞术检测淋巴细胞P-糖蛋白、罗丹明123以及联合甲泼尼龙干预的细胞凋亡率.采用方差分析或t检验进行统计学分析.结果 SLE患者外周血淋巴细胞P-糖蛋白与SLEDAI呈正相关(r=0.490,P<0.05),缓解组P-糖蛋白(12.2±2.5)%与部分缓解或无效组(16.5±4.0)%差异有统计学意义.三七皂苷干预组P-糖蛋白(11.2±3.1)%和罗丹明123(70.1±5.8)%与对照组[分别为(15.3±2.9)%,(53.9±5.2)%]比较差异有统计学意义.三七皂苷可协同甲泼尼龙诱导淋巴细胞凋亡.结论 三七皂苷下调P-糖蛋白表达和转运活性的双重作用可能是协同甲泼尼龙诱导淋巴细胞凋亡的重要机制.

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Objective To investigate the mechanism of glucocorticoid resistance in patients with systemic lupus erythematosus(SLE) and the effects of Panax Notoginseng Saponins(PNS) on reversing glucocorticoid resistance in lymphocytes. Methods The relevance between P-glycoprotein (P-gp, %) and SLEDAI integrals or clinical effects was analyzed by linear correlation analysis. The lymphocytes from SLE patients were intervened with PNS or verapamil. P-gp of the lymphocytes and Rhodamine 123 (Rh123, %) accumulated in the lymphocytes were assayed by flow cytometry. The percentage of the apoptosis of the lymphocytes stimulated with PNS or verapamil combined with methylprednisolone was identified by double-tagging with Annexin V and propidium iodide (PI) and detected by flow cytometry. Variance analysis or Student's t test was used for statistics. Results The P-gp in the peripheral blood lymphocytes of SLE patients was significantly related to SLEDAI integrals. The difference of P-gp between lymphocytes in the completely remission cases (12.2±2.5)% and lymphocytes in the partial remission or non-effective cases (16.5±4.0)% was statistically significantce. The difference of P-gp and Rh123 between lymphocytes treated with PNS [(11.2±3.1)%,(70.1 ±5.8)% respectively ] and lymphocytes of the control group [ (15.3±2.9)%, (53.9±5.2)% respectively ]was statistically significant. The lymphocytes apoptosis rate in the lymphocytes stimulated with methylprednisone combined with PNS increased significantly compared to the lymphocytes stimulated with methylprednisolone only. Conclusion The action of PNS in suppressing both the expression and the transfer activity of P-gp is possibly the important mechanism to increase the effects of methylprednisolone in inducing lymphocytes apoptosis.     

Chronic hepatitis B serum promotes apoptotic damage in human renal tubular cells

AIM: To investigate the effect of the serum of patients with chronic hepatitis B (CHB) on apoptosis of renal tubular epithelial cells in vitro and to study the role of hepatitis B virus (HBV) and transforming growth factor-β1 (TGF-β1) in the pathogenesis of hepatitis B virus associated glomerulonephritis (HBV-GN). METHODS: The levels of serum TGF-β1 were measured by specific enzyme linked immunosorbent assay (ELISA) and HBV DNA was tested by polymerase chain reaction (PCR) in 44 patients with CHB ,and 20 healthy persons as the control. The normal human kidney proximal tubular cell (HK-2) was cultured together with the sera of healthy persons, CHB patients with HBV-DNA nega-tive(20 cases) and HBV-DNA positive (24 cases) for up to 72 h. Apoptosis and Fas expression of the HK-2 were detected by flow cytometer. RESULTS: The apoptosis rate and Fas expression of HK-2 cells were significantly higher in HBV DNA positive serum group 19.01±5.85% and 17.58±8.35%, HBV DNA negative serum group 8.12±2.80% and 6.96±2.76% than those in control group 4.25±0.65% and 2.33±1.09%, respectively (P < 0.01). The apoptosis rate and Fas expression of HK-2 in HBV DNA positive serum group was significantly higher than those in HBV DNA negative serum (P < 0.01). Apoptosis rate of HK-2 cells in HBV DNA positive serum group was positively correlated with the level of HBV-DNA (r = 0.657). The level of serum TGF-β1 in CHB group was 163.05±91.35μg/L, significantly higher as compared with 81.40±40.75μg/L in the control group (P < 0.01). CONCLUSION: The serum of patients with chronic hepatitis B promotes apoptotic damage in human renal tubular cells by triggering a pathway of Fas up-regula-tion. HBV and TGF-β1 may play important roles in the mechanism of hepatitis B virus associated glomerulone-phritis.     

系统性红斑狼疮患者外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的变化及糖皮质激素对其的影响

目的 研究系统性红斑狼疮(SLE)患者外周血CD4+CXCR5+T细胞占CD4+T细胞百分率以及糖皮质激素对其的影响,探讨其在SLE发病机制中的作用.方法 采用流式细胞术检测45例活动期、20例缓解期SLE患者及20名健康对照外周血中CD4+CXCR5+T细胞占CD4+T细胞的百分率,比较其在各组中的差异及糖皮质激素治疗对其的影响,同时检测各组中CD19+B细胞上CXCR5的表达.2组间比较用独立样本t检验,3组间比较采用多变量方差分析,与临床指标之间的相关性分析采用非参数的Spearman相关分析,治疗前后的差异用重复测量的方差分析.结果 ①SLE患者外周血CD4+CXCR5+T细胞占CD4+T细胞的比例高于健康对照组[(16±7)%与(12±3)%,P＜0.01],其中活动组[(18±7)%]高于健康对照组(P＜0.05),而缓解组[(11±4)%]和健康对照组之间差异无统计学意义(P＜0.05);狼疮肾炎组高于非狼疮肾炎组,但差异无统计学意义[(18±7)%与(14±7)%,P=0.05].②CD4+CXCR5+T细胞百分率与SLE疾病活动指数(SLEDAI)、抗核抗体滴度和红细胞沉降率(ESR)呈正相关,与补体C3呈负相关(P均＜0.05),与C反应蛋白、病程、免疫球蛋白无相关性(P＞0.05).抗双链DNA抗体升高组与正常组之间、抗sm抗体、抗SSMSSB抗体阴性组和阳性组之间差异无统计学意义(P均＞0.05).③活动期SLE患者CD19+B细胞上CXCR5的表达比例低于健康对照组[(85±11)%与(94±3)%,P＜0.05].④10例初发、未接受治疗的活动期患者在接受地塞米松(20 mg/d)治疗后第1、3、7天外周血中CD4+CXCR5+T细胞百分率均低于治疗前(P均＜0.05).治疗前后CD19+CXCR5+B细胞的百分率无变化(P均＞0.05).结论 外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的异常町能参与SLE的发病.

Abstract：

Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.     

Children with celiac disease and high tTGA are genetically and phenotypically different

AIM:To investigate whether celiac disease(CD)patients with tissue-transglutaminase antibody(tTGA)≥100 U/mL are different from patients with lower tTGA levels.METHODS:Biopsy-proven(MarshⅢ)pediatric CD patients(n=116)were prospectively included between March 2009 and October 2012.The biopsies were evaluated by a single pathologist who was blinded to all of the patients’clinical data.The patients were distributed into 2 groups according to their tTGA level,which was measured using enzyme-linked immunoassay:tTGA≥100 U/mL and Ttga<100 U/mL.The patients’characteristics,symptoms,human leukocyte antigen(HLA)genotype and degree of histological involvement were compared between the 2 groups.RESULTS:A total of 34(29.3%)children had tTGA values<100 U/mL and 82(70.7%)tTGA levels of≥100 U/mL.Patients with high tTGA levels had lower average body weight-for-height standard deviation scores(SDS)than did patients with tTGA<100 U/mL(-0.20±1.19 SDS vs 0.23±1.03 SDS,P=0.025).In the low tTGA group,gastrointestinal symptoms were more common(97.1%vs 75.6%,P=0.006).More specifically,abdominal pain(76.5%vs 51.2%;P=0.012)and nausea(17.6%vs 3.7%,P=0.018)were more frequent among patients with low tTGA.In contrast,patients with solely extraintestinal manifestations were only present in the high tTGA group(18.3%,P=0.005).These patients more commonly presented with aphthous stomatitis(15.9%vs 0.0%,P=0.010)and anemia(32.9%vs 11.8%,P=0.019).In addition,when evaluating the number of CD-associated HLA-DQ heterodimers(HLA-DQ2.5,HLA-DQ2.2 and HLA-DQ8),patients with low tTGA levels more commonly had only1 disease-associated heterodimer(61.8%vs 31.7%,P=0.005),while patients with high tTGA more commonly had multiple heterodimers.Finally,patients with tTGA≥100 U/mL more often had a MarshⅢc lesion(73.2%vs 20.6%,P≤0.001)while in patients with low tTGA patchy lesions were more common(42.4%vs6.8%,P≤0.001).CONCLUSION:Patients with tTGA≥100 U/mL show several signs of more advanced disease.They also carry a larger number of CD     

系统性红斑狼疮患者外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的变化及糖皮质激素对其的影响

Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.     

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.     

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.     

Fas/FasL途径在氟致人神经母细胞瘤SH-SY5Y细胞凋亡中的作用

目的 探讨Fas/FasL途径在氟致人神经母细胞瘤SH-SY5Y细胞凋亡中的作用.方法 用不同剂量氟化钠[NaF,0(对照)、20、40、80 mg/L].SH-SY5Y细胞进行染毒,24 h后检测细胞存活率、凋亡率和Fas、FasL mRNA表达;选择40 mg/L NaF组,观察在Fas受体激动剂(CH11)或拮抗剂(ZB4)作用下细胞凋亡率及Fas、FasL mRNA表达水平的改变.结果 40、80 mg/L组细胞存活率[(84.63±2.57)%、(69.04±5.63)%]明显低于对照组(100.00%),组间比较差异有统计学意义(P<0.01);细胞凋亡率随染毒剂量的升高呈上升趋势.40、80 mg/L组细胞凋亡率[(8.54±1.95)%、(17.94±2.71)%]明显高于对照组[(3.32±1.33)%],组间比较差异有统计学意义(Jp<0.05);NaF能不同程度地上调Fas、FasL mRNA表达,使Fas/β-actin[40mg/L组(0.94±0.51)、80 mg/L组(0.99±0.12)]和FasL/β-actin[40 mg/L组(0.96±0.42)、80 mg/L组(0.99±0.24)]比值增加.与对照组[Fas/β-actin(0.50±0.33)、FasL/β-actin(0.58±0.23)]比较,差异均有统计学意义(P<0.05).在对细胞凋亡率和Fas、FasL mRNA表达水平的影响中,NaF与CH11之间存在协同作用(,值分别为32.89、18.46、14.69,P<0.01),NaF与ZB4之间存在拈抗作用(F值分别为5.73、24.26、10.17,P<0.05或<0.01).结论 NaF可诱导SH-SY5Y细胞凋亡,Fas/FasL途径在NaF诱导SH-SY5Y细胞凋亡中起重要作用.     

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.     

The pathway of estradiol-induced apoptosis in patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a disease with unknown etiology. The pathologic role of sex hormones and apoptosis in SLE has often been discussed. We studied the effects of estradiol in the pathway of induced apoptosis in Iranian SLE patients. T lymphocytes from 35 SLE patients and 20 age-matched controls were isolated and cultured in the presence of 10⁻⁸ M 17-β estradiol. The expression levels of Fas, Fas ligand (FasL), Bcl-2, caspase-8, and caspase-9 mRNAs were determined semiquantitatively in comparison to the expression level of beta actin RNA. Estradiol exposure did not have any significant effects on the expression levels of Fas, Bcl-2, and caspase-9 in SLE patients and controls. However, the expression levels of FasL and caspase-8 were significantly increased in SLE patients, but not in controls. This suggests the probable involvement of extrinsic apoptosis pathway in estradiol-induced apoptosis in SLE.     

粒细胞集落刺激因子影响冠状动脉微栓塞后心肌细胞凋亡的研究

目的 探讨粒细胞集落刺激因子对大鼠冠状动脉(冠脉)微栓塞后心肌细胞凋亡及左心室功能的影响. 方法 68只雄性SD大鼠,随机分成微栓塞组24只、粒细胞集落刺激因子组24只和假手术组20只.微栓塞组和粒细胞集落刺激因子组升主动脉夹闭后自左心室腔内注入自体微血栓,造成冠脉微栓塞,假手术组注入等量生理盐水.粒细胞集落刺激因子组术后2 h起给予皮下注射重组人粒细胞集落刺激因子100 μg·kg-1·d-1,持续5 d,其余两组给予生理盐水.术后3 d、1周、2周及4周处死大鼠.各组心肌样品中以实时定量聚合酶链式反应法(real time PCR)检测Bcl-2、Bax、Fas及FasL的mRNA表达,并计算Bcl-2/Bax比值,以Western blot蛋白印迹法检测Caspase-3活性及裂解多聚二磷酸腺苷-核糖聚合酶(PARP)蛋白表达水平,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测凋亡细胞及有创压力传感器记录左心室血流动力学改变. 结果 与假手术组比较,微栓塞组术后Bel-2、Bax、Fas及FasL的mRNA表达均有不同程度升高,Bcl-2/Bax比值降低(0.28±0.04和2.98±0.49),Caspase-3及裂解PARP蛋白表达增强(0.762±0.129和0.133±0.027;0.992±0.146和0.386±0.074),心肌细胞凋亡指数升高(17.2±1.9和1.2±0.6),左心室收缩压(LVSP)及左心室内压最大上升和下降速率(±dp/dtmax)明显降低(P<0.05或P<0.01);与微栓塞组比较,粒细胞集落刺激因子组术后Bcl-2的mRNA表达增强、Bax、Fas及FasL的mRNA表达均有不同程度降低,Bcl-2/Bax比值升高(2.07±0.29和0.28±0.04),aspase-3及裂解PARP蛋白表达减弱(0.371±0.041和0.762±0.129;0.548±0.093和0.992±0.146),心肌细胞凋亡指数降低(6.1±1.0和17.2±1.9),LVSP及±dp/dtmax明显升高(P<0.05或P<0.01). 结论 徽栓塞可导致心肌细胞凋亡,降低左心室功能;粒细胞集落刺激因子通过减轻微栓塞后心肌细胞凋亡,改善左心室功能.     

IκB激酶复合物对系统性红斑狼疮患者骨髓间充质干细胞迁移及增殖的作用

目的 观察系统性红斑狼疮(SLE)患者骨髓间充质干细胞(BMSCs)迁移、增殖变化,并探讨IKB激酶复合物(IKK-β)对SLE患者BMSCs迁移及增殖的影响.方法 采用密度梯度离心和贴壁分离法分离培养6例SLE患者及6名健康人BMSCs；通过细胞划痕实验和构建侵袭小室检测BMSCs迁移、活细胞计数(CCK-8)检测BMSCs增殖能力；实时荧光定量聚合酶链反应技术检测IKK-β mRNA表达水平；蛋白印迹法检测IKK-β、磷酸化IKK-β表达水平；观察加入IKK-β抑制剂对SLE患者BMSCs增殖、迁移的影响.采用t检验或Mann-Whitney秩和检验.结果 ①体外培养过程中,SLE患者BMSCs迁移率(5.2±3.8)‰与增殖能力(0.21±0.49)显著低于健康对照组BMSCs迁移率(7.0±2.9)‰及增殖能力(1.00±0.35),差异有统计学意义(P＜0.05);②SLE患者BMSCs IKK-β基因表达水平(1.9±1.4)与健康人(1.9±2.4)相比差异无统计学意义(P＞0.05)；SLE患者BMSCs磷酸化IKK-β蛋白表达水平(1.41±0.19)显著高于健康人(0.93±1.24),差异有统计学意义(P＜0.05)；③IKK-β抑制剂能够增加SLE患者BMSCs的迁移率(3.3±1.6)‰和增殖能力(1.13±0.26),高于未加IKK-β抑制剂组迁移率(2.3±1.1)‰与增殖能力(0.8l±0.17),差异有统计学意义(P＜0.05).结论 SLE患者BMSCs在体外培养过程中,迁移、增殖能力较健康人显著降低,其机制可能是SLE患者体内IKK-β的表达增加,通过活化核因子-KB信号通路,影响迁移与增殖相关基因的表达,从而抑制了BMSCs的迁移与增殖.     

老年大鼠T细胞凋亡相关基因表达模式的研究

目的　探讨老年大鼠 T细胞凋亡的基因表达模式。方法　本研究利用 TUNEL标记的流式细胞检测和荧光实时定量 RT- PCR技术 ,研究了老年大鼠和年轻大鼠 T淋巴细胞凋亡情况及抗凋亡和促凋亡基因 (Fas,Fas L ,bcl- 2 ,bax,TNFR1 ,TNFR2 )的表达差异 ,以及 Caspase8、Caspase3活性的变化。结果　与年轻大鼠相比 ,老年大鼠激活诱导的细胞凋亡百分率增高 ,有显著性差异 (P<0 .0 1 ) ;老年大鼠促凋亡的 Fas,Fas L,TNFR1基因表达上调 ,抗凋亡的 bcl- 2 ,TNFR2基因表达下调 ,与年轻大鼠相比均有显著性差异 (P<0 .0 5或 P<0 .0 1 )。此外 ,老年大鼠激活诱导的T淋巴细胞 Caspase8、3活性增高 ,与年轻大鼠相比有显著性差异 (P<0 .0 1 )。结论　激活诱导的 T细胞过度凋亡与衰老密切相关 ;促凋亡基因表达上调及抗凋亡基因表达下调及 Caspase8、3活性增高是老年大鼠 T细胞凋亡易感性增高重要分子机制。     

骨髓间充质干细胞移植对脑缺血大鼠脑组织凋亡相关蛋白的影响

目的 探讨经静脉移植的骨髓间充质干细胞(MSCs)在脑缺血大鼠脑组织的分布情况及对凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3(Caspase 3)及Bcl-2蛋白表达的影响.方法 体外培养及扩增MSCs后,用绿色荧光染料羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记,通过静脉途径移植给大脑中动脉缺血2 h再灌注的SD大鼠,按不同时间点取材,荧光显微镜下观察MSCs在脑内的分布,免疫组化染色检测大鼠脑内Caspase 3、Bcl-2蛋白表达情况.结果 移植组6、12、24、72 h及7 d时,Caspase 3免疫组化阳性目标面密度分别为(2.81±0.35)%、(3.98±0.67)%、(5.58±0.92)%、(3.51±0.63)%、(1.64±0.29)%,明显低于对照组[(3.92±0.44)%、(5.23±0.30)%、(6.89±0.57)%、(4.39±0.57)%、(2.29±0.21)%],两组比较差异有统计学意义(t值分别为4.37、3.34、2.60、2.32、3.90,P＜0.05或＜0.01);移植组6、12 h及7 d,Bcl-2阳性目标面密度分别为(4.70±0.16)%、(5.61±0.26)%、(3.00±0.28)%,明显高于对照组[(3.28±0.27)%、(4.54±0.59)%、(2.15±0.62)%],两组比较差异有统计学意义(t值分别为8.32、3.25、2.54,P＜0.05或＜0.01).结论 MSCs可能通过下调Caspase 3蛋白表达、上调Bcl-2蛋白表达的方式对脑缺血再灌注损伤起保护作用.     

自体骨髓单个核细胞移植治疗扩张型心肌病疗效观察

目的 观察扩张型心肌病患者经冠状动脉自体骨髓单个核细胞移植治疗的安全性及近中期疗效.方法 258例扩张型心肌病患者,传统治疗的基础上,根据是否行经冠状动脉自体骨髓单个核细胞移植分为移植组(n=71)和对照组(n=187).随访两组患者术前、术后1、3、6个月和1、2年超声心动图、动态心电图、6 min步行距离及心脏核素(SPECT)检查,记录年住院天数.结果 移植组6 min步行距离术后1个月明显优于对照组[(345±76)m比(286±104)m,P<0.05].术后各随访时间点比较,移植组均优于对照组(P<0.05).术后1个月,移植组左心窜射血分数(LVEF)明显高于对照组[(41.5±9.4)%比(37.3±6.6)%,P<0.05].术后2年时移植组LVEF略高于对照组,差异无统计学意义[(43.6±6.3)%比(43.2±6.0)%,P>0.05].术后3个月移植组缺血心肌节段数较术前减少,且较对照组少[分别为(2.0±1.0)个比(3.1±1.4)个和(2.0±1.0)个比(3.1±1.2)个,P均<0.05],而坏死心肌节段数移植前后无明显改变.两组患者牛存率比较,差异无统计学意义(95.4%比94.9%,P>0.05).但移植组患者年住院天数明显少于对照组[(23.6±13.4)d比(33.0±14.0)d,P<0.05].结论 经冠状动脉自体骨髓单个核细胞移植具有良好的安全性,能够提高扩张型心肌病患者的LVEF、增加6 min步行距离,减少年住院大数,且近期疗效显著,中期疗效与传统治疗相似.