【作者】 巩琰；

【导师】 魏世辉；

【作者基本信息】 中国人民解放军军医进修学院 ， 眼科学， 2008， 硕士

【摘要】 目的:通过检验不同发病阶段Eales病患者血液中Th1、Th2细胞因子、氧化及抗氧化水平的变化,探讨免疫平衡、氧化及抗氧化损伤机制在Eales病发病过程中的作用,为该病的诊断、治疗及预后判定提供试验参考及理论依据。方法:1.标本选择及处理:纳入符合Eales病诊断标准的30例患者,系首诊于解放军总医院眼科的Eales病人,全部为男性。最小年龄18岁,最大年龄45岁,平均年龄为28.47±8.03岁;随访时间为3-15年,平均为7.1年,其中:NVH组12例,全部为男性,发病年龄为19-41岁,平均为30.5±8.72岁;VH组18例,发病年龄为18-45岁,平均为26.5±12.53岁。对照组10例,均系本院工作人员。选择范围:男性,年龄在18—45岁之间,无自身免疫病及遗传病家族史,无亲缘关系,全身及眼部检查正常。糖皮质激素治疗组:共16例,男性。最小年龄24岁,最大年龄38岁,平均年龄为27.29±4.67岁;其中NVH组激素治疗患者为10例,VH组激素治疗患者为6例,随访时间为3个月。入选病例清晨7时安静、空腹状态下抽取肘静脉血约4ml,1小时内离心(4℃,3000rpm,10min),用微量移液器吸出上层血清约0.5ml,分装于Eppendorf管中,编号,-70℃保存备用,禁止反复冻融。2.本课题采用多功能液相芯片分析系统(luminex100TM),结合传统ELISA、免疫组化及荧光技术,检测了细胞因子(IL-2、4、5、10,TNF-alpha,IFN-gamma)在不同时期Eales病的水平,分析细胞因子在Eales病患者血清中的含量及其对Eales病的影响,探索Eales病发病因素与发生机制,为减少Eales病的发病率和更好地防治Eales病奠定了一定的基础。3.采用SOD、NO试剂盒分析SOD、NO在Eales病患者血清中的含量,分析氧化、抗氧化失衡对Eales病的影响4.统计分析:用SPSS13.0统计软件分析,P＜0.05认为有统计学差异,所得结果以(?)±S表示,小数点后保留4位有效数字。单因素分析运用T检验分析与Eales病可能有关的危险因素,logistic回归模型进行多因素分析,筛选可能导致Eales病的危险因素及其危险度的大小。结果:1 IFN-gamma:NVH组Eales病人的血清IFN-gamma含量与正常对照组无显著性差异(P≥0.05);VH组Eales病人的血清IFN-gamma含量显著高于正常对照组(P＜0.05);激素治疗病例组Eales病人的血清IFN-gamma含量显著高于正常对照组(P＜0.05];NVH组和VH组Eales病人的血清IFN-gamma含量无显著性差异(P≥0.05);NVH组、VH组Eales病人的血清IFN-gamma含量与激素治疗组无显著性差异(P≥0.05)。2IL-10:NVH组Eales病人的血清IL-10含量与正常对照组无显著性差异(P≥0.05);VHEales病人的血清IL-10含量显著高于正常对照组(P＜0.05);激素治疗病例组Eales病人的血清IL-10含量显著高于正常对照组(P＜0.05);VH组Eales病人的血清IL-10含量显著高于NVH组(P＜0.05);NVH组Eales病人的血清IL-10含量与激素治疗组无显著性差异(P≥0.05);VH组Eales病人的血清IL-10含量显著高于激素治疗组(P＜0.05)。3 IL-2:不同时期Eales病人血液中IL-2含量与正常对照组两两比较均无统计学意义(P≥0.05)。提示IL-2不随Eales病人病情发展而有明显变化。4.IL-5:NVH组Eales病人的血清IL-5含量与正常对照组无显著性差异(P≥0.05);VH组、激素治疗病例组Eales病人的血清IL-5含量均显著高于正常对照组(P＜0.05);VHEales病人的血清IL-5含量显著高于NVH组(P＜0.05);NVH组、VH组Eales病人的血清IL-5含量均显著高于激素治疗组(P＜0.05)。5.IL-4:NVH组、VH组Eales病人的血清IL-4含量低于正常对照组(P＜0.05);激素治疗病例组Eales病人的血清IL-4含量与正常对照组无显著性差异(P≥0.05);VH组Eales病人的血清IL-4含量低于NVH组(P＜0.05);NVH组、VH组Eales病人的血清IL-4含量低于激素治疗组(P＜0.05)。6.TNF-alpha:NVH、VH组Eales病人的血清TNF-alpha含量均显著高于正常对照组(P＜0.05);激素治疗病例组Eales病人的血清TNF-alpha含量与正常对照组无显著性差异(P＜0.05);NVH组Eales病人的血清TNF-alpha含量显著高于VH组(P≥0.05);NVH组、VH组Eales病人的血清TNF-alpha含量均高于激素治疗组(P＜0.05)。7.SOD:不同时期Eales病人的血清SOD含量与正常对照组均有显著性差异(P＜0.05);NVH组Eales病人的血清SOD含量显著高于VH组(P＜0.05);NVH组、VH组Eales病人的血清SOD含量均显著低于激素治疗组(P＜0.05)。8.不同时期Eales病人的血清NO含量与正常对照组均有显著性差异(P＜0.05);激素治疗后血清NO含量与正常对照组差异无统计学意义(P＞0.05)。NVH组与VH组Eales病人的血清NO含量组差异无统计学意义(P＞0.05);NVH组、VH组Eales病人的血清NO含量均显著低于激素治疗组(P＜0.05)。将以上所述的免疫细胞因子赋值后带入多元线性回归模型,得到回归方程Y=6.91+3.22X。假设检验F=11.843,P=0.01,有统计学意义。最终进入回归方程的为TNF-alpha,且Eales病影响依次减弱。结论:1:TH细胞亚群异常在Eales病的发生发展中起着重要作用2:减少Th1型细胞因子的产生,平衡Th1/Th2可减少局部T淋巴细胞聚集,进而可能治疗Eales病和改变病程。3.糖皮质激素治疗Eales病尤其是NVH患者具有显著疗效,但有些细胞因子水平仍偏高,提示疾病仍有复发的可能性。4.氧化应激与Eales病关系密切,清除自由基也可能起到治疗Eales病的功效。更多还原

【Abstract】 Objective:to investigate changes in Th1Th2cytokine,oxidation and antioxidant ingredients in different pathological durations,then infer immune mechanism,role of oxidation and anti-oxidative damage to the Eales disease in the process and evaluateMathod:1.Sample selection and processing:30 Eales disease patients,first diagnosed in the Department of ophthalmology,The PLA General Hospital,all were male.Minimum age of 18-year-old,maximum age 45,The average age of 29.29±8.67 years,follow-up time for 3-15 years,an average of 7.1 years.Among that patients,12 were in NVH period group vs 18 VH patients.The age of 19-41 years old,The average age of 32.5±4.53;VH group of 20 patients,age of onset was 18-45 years old,an average of 26.5±12.53.The control group was 10 cases,all the hospital staff.They all were males,aged between 18 an 45-year-old,no autoimmune diseases and family history of genetic disease,unrelated, normal body and eye examination;Corticosteroid therapy group:a total of 16 cases,male. Minimum age of 24-year-old,maximum age of 38-year-old,The average age of 27.29±4.67 years follow-up time is 3-15 years,with an average of 6.9 years 7:00 am all selected cases were taken blood from The elbow about 4 ml at quiet,fasting state.After centrifugal (4℃,3000 rpm,10min)1 hour,sucked with micro-tips from the upper serum of blood about 0.5 ml,points installed in Eppendorf tube,numbered,-70℃saved standby to prohibit The repeated freezing and Thawing.2.The subject used liquid-chip analysis system (luminex100TM),combining traditional ELISA,immunohistochemistry and fluorescence to detect The cytokines(IL-2,4,5,10,TNF-α,IFN-γ)level in different Eales disease duration.we compared the contents of cytokines in the serum of patients with Eales and its impact on Eales disease,analyse Eales disease’ risk factors and reduce the incidence of Eales disease and to find a better way to prevent and treat the disease.3.Use of SOD,NO kit to analyze the serum content of SOD,NO in The patients and the impact of oxidation, anti-oxidation imbalance of the of Eales disease4.Statistical analysis:T test used to analysis of related risk factors,logistic regression model with multi-factor analysis of risk factors could leading to Eales disease and the risk of size. Result:1.HIFN-γ:the results of the statistical analysis showed that in The inflammation duration,the level of serum HIFN-γwas no significant difference compared with control group(P≥0.05);VH group was significantly higher than control group(P＜0.05);glucocorticosteroid group was also significantly higher than control group(P＜0.05);NVH group and VH group was no significant difference(P≥0.05);NVH group, VH group and glucocorticosteroid cured group were no significant difference(P≥0.05). 2.IL-10:the level of serum IL-10 in NVH group was no significant difference competed with control group(P≥0.05);VH group was significantly higher than control group(P＜0.05);glucocorticosteroid group was significantly higher than the control group(P＜0.05); VH group was significantly higher than NVH group(P＜0.05);NVH Eales group and glucocorticosteroid group were no significant difference(P≥0.05);group of VH was significantly higher than glucocorticosteroid group(P＜0.05).3.IL-2:the blood level of IL-2 patients was no significant change in different periods compared with the control group(P≥0.05).IL-2 did not significant change with the development of disease.4.IL-5: The results of the statistical analysis showed that the NVH phase Eales patient’s serum IL-5 in the control group and no significant difference(P≥0.05);group of VH, glucocorticosteroid cure the patient’s group Eales serum IL-5 levels were significantly higher than the normal control group(P＜0.05);VH phase Eales patient’s serum IL-5 was significantly higher than the NVH phase group(P＜0.05);NVH phase group,VH group of the patient’s serum IL-5 levels were significantly higher than the glucocorticosteroid group (P＜0.05).5.IL-4:the level of serum IL-4 in NVH group was significant lower than the control group(P＜0.05);group of VH was significantly higher than the normal control group(P＜0.05);glucocorticosteroid group was no significant difference with the control group(P≥0.05);VH group was significantly lower than the glucocorticosteroid group (P＜0.05).6.TNF-α:The results of the statistical analysis showed that the serum TNF-αlevels in NVH group,VH group were significantly higher than the control group(P＜0.05); glucocorticosteroid group was no significant difference compared with the control group(P≥0.05);NVH group was significantly higher than VH group(P≥0.05);NVH group were higher than glucocorticosteroid group(P＜0.05).7.SOD results of the statistical analysis showed that level of serum SOD in different periods Eales of patients were significantly different with that the control group(P＜0.05);NVH group was significantly higher than VH group(P＜0.05);NVH group,VH group were significantly lower than glucocorticosteroid group(P＜0.05).8.NO results of the statistical analysis showed that level of serum NO in different periods Eales of patients were significantly different with that in the control group(P＜0.05);after glucocorticosteroid therapy serum content of NO was no significant difference with the control group(P＞0.05).VH and NVH groups were not significant difference(P＞0.05);NVH group,VH group were significantly lower than the glucocorticosteroid group(P＜0.05).Conclusions:1:Th cell subsets plays an important role in Eales disease’development 2:Reducing Th1-produced cytokines and balance the Th1/Th2 T-lymphocytes may possibly treat the Eales disease.3.Glucocorticoid treatment of Eales disease especially in NVH group take a significant effect,but some cytokines are still on a high level,suggesting that there is still a possibility recurrence of Eales disease.4.Oxidative stress is also closely related to Eales disease;free radicals elimination may also play an important role in The treatment of Eales disease.更多还原