转化生长因子β1在干眼患者眼表的表达研究

发布时间：2018-02-24 05:21

本文关键词： 转化生长因子β1 干眼印记细胞 real-time PCR　出处：《山西医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：探讨转化生长因子β1(TGF-β1)在干眼患者眼表的表达,及其与干眼分级、症状、体征的相关性。方法：通过OSDI干眼调查问卷、泪膜破裂时间测定(TBUT)、泪液分泌试验(Schirmer I Test)、眼表染色筛选出2011年4月至12月我院门诊就诊的干眼患者30例(30眼),对照组15例(15眼)为正常人。所有病例及对照组均做如下临床症状及体征监测：OSDI干眼调查问卷,TBUT,泪液分泌试验,角膜荧光素钠染色,结膜丽丝胺绿染色。并按照OSDI评分将干眼病例分为轻度组(18例18眼)和中重度组(12例12眼)。应用印记细胞法收集不同程度干眼患者和正常对照组两组球结膜上皮细胞分别进行LHC法染色和real-time PCR法检测结膜中TGF-β1的基因表达。结果： ①轻度组、中重度组及正常对照组OSDI评分值分别为：22.61±2.20、44.67±7.58、9.00±3.34,不同程度干眼组和正常对照组OSDI评分差异有统计学意义(P0.05)。轻度组、中重度组及正常对照组泪液分泌试验结果分别为：7.33±1.97mm、3.58±1.62mm、10.40±2.69mm,不同程度干眼组和正常对照组泪液分泌试验结果差异有统计学意义(P0.05)。轻度组、中重度组及正常对照组TBUT值分别为：7.50±2.18s、3.58±1.16s、11.20±2.21s,不同程度干眼组和正常对照组TBUT值差异有统计学意义(P0.001)。轻度组、中重度组及正常对照组角膜荧光素钠染色评分值分别为：2.67±1.68、5.83±1.99、0.60±0.51,不同程度干眼组和正常对照组角膜荧光素钠染色评分差异有统计学意义(P0.05)。轻度组、中重度组及正常对照组结膜丽丝胺绿染色评分值分别为：4.44±1.98、6.25±1.96、1.93±1.58,不同程度干眼组和正常对照组结膜丽丝胺绿染色评分差异有统计学意义(P0.05)。 ②轻度组、中重度组及正常对照组TGF-β1相对表达量分别为：1.40±0.20、2.07±0.42、0.73±0.42,不同程度干眼组和正常对照组TGF-β1相对表达量差异有统计学意义(P0.05)。 ③TGF-β1在结膜上皮细胞中的表达率与OSDI评分存在正相关关系(r=0.814,P0.05),与角膜荧光素钠染色评分存在正相关关系(r=0.748,P0.05),与结膜丽丝胺绿染色评分存在正相关关系(r=0.590,P0.05),与泪液分泌试验结果存在负相关关系(r=-0.645,P0.05),与TBUT存在负相关关系(r=-0.693,P0.05)。结论：印迹细胞法取材联合IHC法染色或real-time PCR法检测分析是研究多种眼表疾病发病机制的有效方法之一。免疫组织化学法(IHC)染色可见干眼患者结膜上皮细胞胞质中TGF-β1的表达量较正常对照组明显增多。 real-time PCR法结果显示了干眼患者结膜上皮细胞中TGF-β1的表达量较正常人明显增加。在不同程度干眼患者结膜上皮细胞中TGF-β1的表达率与OSDI干眼调查问卷评分、角膜荧光素钠染色评分及结膜丽丝胺绿染色评分呈正相关,与泪液分泌试验及TBUT呈负相关。因而TGF-β1干眼患者结膜上皮细胞中的表达情况可以反映疾病的严重程度,进而为干眼的诊断和分级提供新的参考依据。
[Abstract]:Objective:. To investigate the expression of transforming growth factor 尾 1 (TGF- 尾 1) in dry eye and its correlation with dry eye grade, symptoms and signs. Methods:. Through OSDI dry eye questionnaire, The tear film rupture time was determined by TBUTT, the tear secretion test was Schirmer I test, eye surface staining was used to screen 30 dry eye patients (30 eyes) from April 2011 to December, and 15 eyes (15 eyes) from the control group. All the cases and the control group were all normal. Do the following clinical symptoms and signs monitoring: OSDI dry eye questionnaire TBUTT, tear secretion test, Corneal fluorescein sodium staining, According to the OSDI score, the patients with dry eye were divided into mild group (18 cases, 18 eyes) and moderate and severe group (12 cases, 12 eyes). The bulb conjunctival epithelium was collected by imprint cell method in patients with different degrees of dry eye and normal control group. TGF- 尾 1 gene expression in conjunctiva was detected by LHC staining and real-time PCR assay. Results:. 1the OSDI scores of mild group, moderate severe group and normal control group were respectively: 1. The OSDI scores of mild group, moderate group and control group were respectively: 1. The OSDI score of dry eye group was significantly different from that of normal control group (P 0.05, P < 0.05), and that of normal control group was 44.67 卤7.58 卤9.00 卤3.34, respectively. The results of tear secretion test in moderate and severe group and normal control group were respectively 7. 33 卤1. 97 mm and 3. 58 卤1. 62 mm and 10. 40 卤2. 69 mm, respectively. The difference of tear secretion test between dry eye group and normal control group was statistically significant (P 0. 05). The TBUT values of moderate and severe eyes group and normal control group were 3.58 卤1.16 sg and 11.20 卤2.21 s, respectively. There were significant differences in TBUT between dry eye group and normal control group (P 0.001). The corneal fluorescein sodium staining scores in moderate and severe group and normal control group were: 1: 2.67 卤1.68 卤5.83 卤1.99 卤0.60 卤0.51, respectively. There was a significant difference in corneal fluorescein sodium staining score between the dry eye group and the normal control group (P 0.05). The score of conjunctival riminine green staining in moderate and severe group and normal control group was 4.44 卤1.98 卤6.25 卤1.96 卤1.93 卤1.58, respectively. There was significant difference between the dry eye group and the normal control group (P 0.05). 2the relative expression of TGF- 尾 1 in mild group, moderate severe group and normal control group was 1.40 卤0.202.07 卤0.42 卤0.73 卤0.42, respectively. There was significant difference in the relative expression of TGF- 尾 1 between dry eye group and normal control group (P 0.05). 3There was a positive correlation between the expression rate of TGF- 尾 1 in conjunctival epithelial cells and the OSDI score. There was a positive correlation between the expression rate of TGF- 尾 1 in conjunctival epithelial cells and OSDI score. There was a positive correlation between TGF- 尾 1 expression rate and corneal fluorescein staining score. There was a positive correlation between the expression rate of TGF- 尾 1 in conjunctival epithelial cells and OSDI score, and there was a positive correlation between TGF- 尾 1 expression and corneal fluorescein staining score. The results showed that there was a negative correlation between TBUT and TBUT. Conclusion:. Imprinted cell method combined with IHC staining or real-time PCR assay is one of the effective methods to study the pathogenesis of various ocular surface diseases. Immunohistochemical staining shows TGF- 尾 1 in the cytoplasm of conjunctival epithelial cells in patients with dry eye. The expression level was significantly higher than that in the normal control group. The results of real-time PCR showed that the expression of TGF- 尾 1 in conjunctival epithelial cells of dry eye patients was significantly higher than that of normal subjects. The expression rate of TGF- 尾 1 in conjunctival epithelial cells of dry eye patients with different degrees and the score of OSDI dry eye questionnaire were significantly increased. Corneal fluorescein sodium staining score and conjunctival riminine green staining score were positively correlated, and negatively correlated with tear secretion test and TBUT. Therefore, the expression of TGF- 尾 1 in conjunctival epithelial cells could reflect the severity of the disease. It provides a new reference for the diagnosis and grading of dry eye.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R777

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