Graves眼病患者检测血清VEGF、HA、bFGF、sICAM-1的临床意义及免疫治疗研究

发布时间：2018-07-08 15:14

本文选题：Graves眼病 + 眼眶成纤维细胞　；参考：《南京大学》2011年硕士论文

【摘要】：[背景]： Graves眼病(Graves'ophthalmopathy, GO)是一种慢性自身免疫相关性疾病,由于眼眶球后组织的炎症反应和亲水性氨基葡聚糖的蓄积,伴或不伴有眼眶脂肪组织和眼外肌的增生,引起眼眶球后组织的容积增加导致一系列临床表现,如眼痛、眼球突出、眶周水肿、眼外肌运动障碍、炎症及损害视力等。大约有20～25%的甲状腺机能亢进的病人发生Graves眼病。Graves眼病发病的机制至今仍不是十分清楚,临床治疗也有一定困难。 Graves眼病的病程经历两个阶段：早期为活动期,主要病理表现为眼部淋巴细胞和炎性细胞的浸润、氨基葡聚糖的沉积与水肿；晚期为稳定期,表现为球后组织的纤维化及脂肪的沉积。病程不同时期GO患者采取的治疗方法和疗效不同,因此正确评判Graves眼病的病情对治疗方法的选择和预后的估计有重要意义。国内外文献报道,GO患者血清透明质酸(Hyaluronic Acid, HA)和细胞间粘附分子-1(Soluble intercellular adhesion molecule-1, sICAM-1)水平可作为评价眼眶活动度与严重程度的重要指标,血清碱性成纤维细胞生长因子(Basic fibroblast growth factor, bFGF)和血管内皮生长因子(Vascular endothelial growth factor, VEGF)水平在活动性GO患者中表达增加。目前Graves眼病的免疫治疗,最基本最常用的是糖皮质激素的应用。大量证据表明,糖皮质激素对中、重度浸润性突眼的治疗尤其有效,中、重度及活动性GO患者静脉使用糖皮质激素类药物的有效率可达77%。其他免疫抑制剂包括雷公藤多甙、霉酚酸酯、环孢菌素等在临床都有一定的应用,并且收到了较好的治疗效果。大量循症医学证据表明雷公藤多甙治疗甲亢突眼有效,对细胞免疫及体液免疫均有较强的抑制作用,抑制了眼球后组织自身抗体形成,减轻突眼症状。我们前期的研究发现霉酚酸酯和雷公藤治疗GO的疗效较环孢素好,且不良反应少,在临床上可能有较好的应用前景。 [目的]：通过检测GO患者血清中VEGF、HA、bFGF、sICAM-1四种标记物的水平,寻找能够评价GO患者病情活动性的客观指标,从而更好地评价患者的病情以指导临床治疗。比较GO患者在使用糖皮质激素、雷公藤多甙和霉芬酸脂治疗前后四种因子水平变化,寻求更为有效的治疗药物,并为临床治疗推广提供新的选择。 [方法]： 1、采用ELISA方法检测50例GO患者和31例单纯Graves病无眼征的患者血清中bFGF、HA、sICAM-1、VEGF因子的表达水平,比较两组间四种因子水平的变化,并对GO组患者血清四种因子水平、临床活动性评分及其他指标进行相关性分析。根据临床评分系统将GO患者分为活动组与稳定组,并比较两组间四种因子水平的变化。 2、采用ELISA方法检测12例GO患者血清HA、bFGF、sICAM-1、VEGF因子水平,并对GO患者进行临床活动性评分,分析比较这些因子水平和临床活动性评分在免疫抑制治疗前后的变化,12例患者中口服雷公藤多甙者4例(20mg,3次/d),霉酚酸酯者4例(0.5g,2次/d),强的松者4例(40mg/d×4w,20mg/d×4w, 10mg/d×4w),疗程12周。 3、应用SPSS16.0统计学软件分析,计量资料使用均数±标准差(x±s)表示,并对数据进行正态性检验及方差分析。组间比较采用两独立样本t检验；GO患者治疗前后两组间的比较采用配对t检验；相关性分析采用Spearman相关性分析,P0.05为差异有统计学意义。 [结果]： 1)GO患者血清VEGF、HA、sICAM-1和bFGF水平较Graves病患者明显升高,有统计学差异(p0.05)。GO患者血清四种因子水平与临床积分显著相关(p0.05),与年龄和病程不相关(p0.05)。 2)活动组GO患者血清sICAM-1较稳定组明显升高,有统计学差异(p0.05),而HA和VEGF及bFGF水平较稳定组虽有升高,但无统计学差异(p0.05)。 3)GO患者血清HA、VEGF、bFGF三种因子之间呈显著正相关性(p0.05),而sICAM-1水平与VEGF、bFGF和HA水平均无明显相关性(p0.05)。 4)治疗12周后,各组患者均无眼病恶化情况。GO组经免疫抑制治疗后的血清HA、sICAM-1、bFGF、VEGF水平与治疗前相比均显著降低,且临床活动性评分也显著降低,差异有统计学意义(p0.05),并且83.33%(10/12)的患者在使用免疫抑制剂治疗后临床活动性分值降低3分以上,口服霉酚酸酯和雷公藤各1例患者临床积分降低2分。强的松组4例患者中2例出现高血糖反应和胃肠道反应。MMF和TⅡ组与强的松组相比,无明显的药物不良反应。 [结论]： 1)HA、sICAM-1、VEGF和bFGF在Graves眼病的发病过程中可能相互作用,共同参与GO的发生发展,其血清中的水平与GO病情的活动性密切相关,可作为评价Graves眼病病情活动性的指标。 2)实验结果显示GO患者经免疫抑制药物治疗后血清VEGF、HA、bFGF、sICAM-1水平明显下降,这四种标记物可以作为监测GO病情和调整GO治疗方法的有用指标。
[Abstract]:[background]:
Graves (Graves'ophthalmopathy, GO) is a chronic autoimmune related disease, due to the inflammatory reaction of the orbital tissue and the accumulation of hydrophilic glucan, with or without the proliferation of orbital adipose tissue and extraocular muscles. The increase in the volume of posterior orbital tissue leads to a series of clinical manifestations, such as eye pain, and the eyeball process. Edema, dyskinesia of the extraocular muscles, inflammation and impairment of vision, about 20 to 25% of the hyperthyroidism patients with Graves eye disease.Graves eye disease pathogenesis is still not very clear, and clinical treatment is also difficult.
The course of Graves's ophthalmopathy experienced two stages: early stage of activity, the main pathological manifestations were infiltration of ocular lymphocytes and inflammatory cells, deposition and edema of glucan; late stable period, manifested as the fibrosis of the tissue and the deposition of fat. The treatment and effect of GO patients in the course of the disease were different because of different therapeutic methods and effects. It is important to correctly judge the condition of Graves eye disease for the choice of treatment and the estimation of prognosis. The domestic and foreign literature reports that the level of serum hyaluronic acid (Hyaluronic Acid, HA) and intercellular adhesion molecule -1 (Soluble intercellular adhesion molecule-1, sICAM-1) in GO patients can be used as the evaluation of orbital activity and severity. The levels of serum basic fibroblast growth factor (Basic fibroblast growth factor, bFGF) and vascular endothelial growth factor (Vascular endothelial growth factor, VEGF) were increased in active GO patients.
At present, the most basic and most commonly used immunotherapy for Graves eye diseases is the use of glucocorticoids. A large number of evidence suggests that glucocorticoid is especially effective in the treatment of severe invasive exophthalmos, and the effective rate of intravenous glucocorticoids in the moderate, severe and active GO patients can reach 77%. other immunosuppressants including Tripterygium wilfordii, Mycophenolate mofetil and cyclosporin have been used in clinical practice and have received good therapeutic effects. A large number of evidence-based medical evidence shows that Tripterygium wilfordii polysaccharide is effective in the treatment of hyperthyroidism and has strong inhibitory effect on cellular immunity and humoral immunity. It inhibits the formation of autoantibodies in the post eyeball group and alleviated the symptoms of exophthalmos. The study found that the efficacy of mycophenolate mofetil and Tripterygium Wilfordii in the treatment of GO is better than cyclosporine, and has fewer adverse reactions.
[Objective]:
By detecting the level of four markers of VEGF, HA, bFGF, and sICAM-1 in the serum of GO patients, the objective indicators that can evaluate the activity of the patients with GO are searched for better evaluation of the patient's condition to guide the clinical treatment. The changes of the levels of the GO patients before and after the use of glucocorticoid, Tripterygium Wilfordii and mycophenopinic acid are compared. Seek more effective treatment drugs, and provide new options for clinical treatment and promotion.
[method]:
1, the serum levels of bFGF, HA, sICAM-1, VEGF in the serum of 50 patients with GO and 31 cases of simple Graves disease were detected by ELISA, and the levels of four factors between the two groups were compared, and the levels of four serum factors, clinical activity scores and other indexes were analyzed. The clinical scoring system was based on the clinical scoring system. GO patients were divided into active group and stable group, and the levels of four factors were compared between the two groups.
2, the serum levels of HA, bFGF, sICAM-1 and VEGF were detected by ELISA in 12 patients with GO, and the clinical activity score of patients with GO was evaluated. The changes of these factors and clinical activity scores before and after the immunosuppressive therapy were compared. 4 cases (20mg, 3 /d), 4 cases of mycophenolate mofetil (0.5g, 2 /d) were taken in 12 patients with the changes of these factors and clinical activity scores before and after the immunosuppressive therapy. Prednisone was found in 4 patients (40mg/d x 4W, 20mg/d x 4W, 10mg/d x 4W) for 12 weeks.
3, using SPSS16.0 statistical software analysis, the measurement data were expressed with mean standard deviation (x + s) and normal test and variance analysis of the data. Two independent sample t tests were used in the group. The comparison of the two groups before and after the treatment of GO was compared with the paired t test; the correlation analysis adopted the Spearman correlation analysis and the P0.05 was the difference. There is a statistical significance.
[results]:
1) the levels of serum VEGF, HA, sICAM-1 and bFGF in patients with GO were significantly higher than those of Graves patients. There was a statistically significant difference (P0.05) the level of four factors in serum of.GO patients was significantly related to clinical integration (P0.05), not related to age and course of disease (P0.05).
2) the serum sICAM-1 of GO patients in the active group was significantly higher than that in the stable group (P0.05), while the levels of HA and VEGF and bFGF were higher than those in the stable group, but there was no statistical difference (P0.05).
3) there was a significant positive correlation between the three factors of serum HA, VEGF and bFGF in GO patients (P0.05), while sICAM-1 level had no significant correlation with VEGF, bFGF and HA levels (P0.05).
4) after 12 weeks of treatment, the patients in each group had no worsening of eye disease. The serum HA, sICAM-1, bFGF, and VEGF levels in the.GO group were significantly lower than those before the treatment, and the clinical activity score was significantly lower, and the difference was statistically significant (P0.05), and 83.33% (10/12) patients were used in clinical activities after the use of immunosuppressive therapy. The score of sex score was reduced by more than 3, and the clinical integral of 1 patients with oral mycophenolate mofetil and Tripterygium wilfordii was reduced by 2. 2 of the 4 patients in the prednisone group had hyperglycemia and gastrointestinal reaction.MMF and T II, and there was no obvious adverse drug reaction compared with the prednisone group.
[Conclusion]:
1) HA, sICAM-1, VEGF and bFGF may interact in the pathogenesis of Graves ophthalmopathy, and participate in the development of GO. The level of serum is closely related to the activity of GO, and can be used as an index to evaluate the activity of Graves eye disease.
2) the experimental results showed that the levels of serum VEGF, HA, bFGF and sICAM-1 decreased significantly after the treatment of immunosuppressive drugs in GO patients. These four markers could be used as a useful indicator to monitor the condition of GO and to adjust the treatment of GO.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R581;R771.3

【参考文献】