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  本文關鍵詞： 轉化生長因子β1 干眼 印記細胞 real-time PCR　出處：《山西醫(yī)科大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的： 探討轉化生長因子β1(TGF-β1)在干眼患者眼表的表達,及其與干眼分級、癥狀、體征的相關性。 方法： 通過OSDI干眼調查問卷、淚膜破裂時間測定(TBUT)、淚液分泌試驗(Schirmer I Test)、眼表染色篩選出2011年4月至12月我院門診就診的干眼患者30例(30眼),對照組15例(15眼)為正常人。所有病例及對照組均做如下臨床癥狀及體征監(jiān)測：OSDI干眼調查問卷,TBUT,淚液分泌試驗,角膜熒光素鈉染色,結膜麗絲胺綠染色。并按照OSDI評分將干眼病例分為輕度組(18例18眼)和中重度組(12例12眼)。應用印記細胞法收集不同程度干眼患者和正常對照組兩組球結膜上皮細胞分別進行LHC法染色和real-time PCR法檢測結膜中TGF-β1的基因表達。 結果： ①輕度組、中重度組及正常對照組OSDI評分值分別為：22.61±2.20、44.67±7.58、9.00±3.34,不同程度干眼組和正常對照組OSDI評分差異有統(tǒng)計學意義(P0.05)。輕度組、中重度組及正常對照組淚液分泌試驗結果分別為：7.33±1.97mm、3.58±1.62mm、10.40±2.69mm,不同程度干眼組和正常對照組淚液分泌試驗結果差異有統(tǒng)計學意義(P0.05)。輕度組、中重度組及正常對照組TBUT值分別為：7.50±2.18s、3.58±1.16s、11.20±2.21s,不同程度干眼組和正常對照組TBUT值差異有統(tǒng)計學意義(P0.001)。輕度組、中重度組及正常對照組角膜熒光素鈉染色評分值分別為：2.67±1.68、5.83±1.99、0.60±0.51,不同程度干眼組和正常對照組角膜熒光素鈉染色評分差異有統(tǒng)計學意義(P0.05)。輕度組、中重度組及正常對照組結膜麗絲胺綠染色評分值分別為：4.44±1.98、6.25±1.96、1.93±1.58,不同程度干眼組和正常對照組結膜麗絲胺綠染色評分差異有統(tǒng)計學意義(P0.05)。 ②輕度組、中重度組及正常對照組TGF-β1相對表達量分別為：1.40±0.20、2.07±0.42、0.73±0.42,不同程度干眼組和正常對照組TGF-β1相對表達量差異有統(tǒng)計學意義(P0.05)。 ③TGF-β1在結膜上皮細胞中的表達率與OSDI評分存在正相關關系(r=0.814,P0.05),與角膜熒光素鈉染色評分存在正相關關系(r=0.748,P0.05),與結膜麗絲胺綠染色評分存在正相關關系(r=0.590,P0.05),與淚液分泌試驗結果存在負相關關系(r=-0.645,P0.05),與TBUT存在負相關關系(r=-0.693,P0.05)。 結論： 印跡細胞法取材聯(lián)合IHC法染色或real-time PCR法檢測分析是研究多種眼表疾病發(fā)病機制的有效方法之一。免疫組織化學法(IHC)染色可見干眼患者結膜上皮細胞胞質中TGF-β1的表達量較正常對照組明顯增多。 real-time PCR法結果顯示了干眼患者結膜上皮細胞中TGF-β1的表達量較正常人明顯增加。在不同程度干眼患者結膜上皮細胞中TGF-β1的表達率與OSDI干眼調查問卷評分、角膜熒光素鈉染色評分及結膜麗絲胺綠染色評分呈正相關,與淚液分泌試驗及TBUT呈負相關。因而TGF-β1干眼患者結膜上皮細胞中的表達情況可以反映疾病的嚴重程度,進而為干眼的診斷和分級提供新的參考依據(jù)。
[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 鹵7.58 鹵9.00 鹵3.34, respectively. The results of tear secretion test in moderate and severe group and normal control group were respectively 7. 33 鹵1. 97 mm and 3. 58 鹵1. 62 mm and 10. 40 鹵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 鹵1.16 sg and 11.20 鹵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 鹵1.68 鹵5.83 鹵1.99 鹵0.60 鹵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 鹵1.98 鹵6.25 鹵1.96 鹵1.93 鹵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 鹵0.202.07 鹵0.42 鹵0.73 鹵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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R777

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本文編號：1529019