首诊干眼患者结膜黏蛋白表达变化及临床意义

Changes in conjunctival mucin expressions in patients with first diagnosis of dry eye and its clinical significance

目的了解首诊干眼患者结膜黏蛋白MUC1、MUC4、MUC5AC和MUC16表达情况及其与干眼症状体征的相关性。方法采用横断面研究方法,于2016年12月至2018年5月在厦门大学附属厦门眼科中心、首都医科大学附属北京同仁医院、四川大学华西医院和上海市普陀区中心医院4个临床研究中心纳入首次就诊干眼患者69例69眼作为干眼组,正常志愿者40例40眼作为正常对照组。受检者均填写中国干眼问卷、眼表疾病指数量表(OSDI)和干眼相关生活质量评分问卷(DEQS),以评估干眼相关症状;采用泪膜破裂时间(TBUT)检查、角结膜荧光素钠染色、基础泪液分泌试验(Schirmer试验Ⅰ)以评估干眼相关体征;采用结膜印迹细胞学方法采集结膜细胞,采用实时荧光定量PCR法检测受检者结膜中MUC1、MUC4、MUC5AC和MUC16 mRNA表达情况;采用Spearman秩相关分析探讨结膜黏蛋白表达量与干眼症状体征之间的关联。结果干眼组患者结膜黏蛋白MUC1和MUC16 mRNA相对表达量分别为3.277(0.568,5.790)和1.815(1.048,3.694),均较正常对照组的1.055(0.550,2.010)和1.024(0.541,1.965)升高,差异均有统计学意义(Z=819.00,P=0.008;Z=861.00,P=0.002)。根据OSDI症状分级,轻中度干眼(评分12-32)以MUC1表达升高为主[3.277(1.161,6.226),Z=9.04,P=0.029],重度干眼(评分>32)主要以MUC16表达升高为主[1.968(1.074,3.726),Z=12.24,P=0.007]。MUC1 mRNA相对表达量与TBUT呈正相关,与角膜染色评分和角结膜染色评分呈负相关(r s=0.270,P=0.025;r s=-0.331,P=0.006;r s=-0.325,P=0.007)。MUC16 mRNA相对表达量与TBUT呈正相关,与视物模糊症状评分、阅读时症状加重评分、开夜车影响程度评分、使用计算机影响程度评分及看电视影响程度评分均呈负相关(r s=0.249,P=0.039;r s=-0.359,P=0.047;r s=-0.370,P=0.034;r s=-0.558,P=0.016;r s=-0.498,P=0.006;r s=-0.515,P=0.002)。结论干眼患者结膜黏蛋白MUC1与MUC16 mRNA表达水平升高,MUC1 mRNA表达与患者体征相关,MUC16 mRNA表达与患者的生活质量相关。

Objective To investigate the expressions of MUC1,MUC4,MUC5AC and MUC16 in patients with first diagnosis of dry eye and their correlation with dry eye symptoms and signs.Methods A cross-sectional study was conducted.Sixty-nine dry eye patients(69 eyes)as dry eye group and 40 normal volunteers(40 eyes)as normal control group were recruited in Xiamen Eye Center of Xiamen University,Beijing Tongren Hospital,West China Hospital of Sichuan University and Shanghai Puotuo District Center Hospital from December 2016 to May 2018.Symptoms were evaluated by Chinese dry eye questionnaire,Ocular Surface Disease Index(OSDI)and Dry Eye-Related Quality-of-Life Score Questionnaire(DEQS).Signs were assessed by tear film breakup time(TBUT),keratoconjunctival fluorescein sodium staining,and Schirmer I test.Conjunctival cells were collected by conjunctival impression cytology.The expression levels of MUC1,MUC4,MUC5AC and MUC16 mRNA in the two groups were determined by real-time fluorescence quantitative PCR.The correlation between the mRNA levels of conjunctival mucins and dry eye symptoms and signs were analyzed by Spearman correlation analysis.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committees of Xiamen Eye Center of Xiamen University(No.2017003),Beijing Tongren Hospital,Capital Medical University(No.TREC2016-29),West China Hospital of Sichuan University(No.2016310)and Shanghai Puotuo District Center Hospital(No.PTEC-A-2016-18-1).Written informed consent was obtained from each subject before any medical examination.Results The expression levels of MUC1 and MUC16 mRNA in dry eye patients were 3.277(0.568,5.790)and 1.815(1.048,3.694),which were higher than 1.055(0.550,2.010)and 1.024(0.541,1.965)in normal control group(Z=819.00,P=0.008;Z=861.00,P=0.002).According to OSDI scores,MUC1 was mainly increased to 3.277(1.161,6.226)in mild to moderate(12-32 points)dry eye patients(Z=9.04,P=0.029),and MUC16 was mainly increased to 1.968(1.074,3.726)in severe(>32 points)dry eye patients(Z=12.24,P=0.007).MUC1 expression was positively correlated with TBUT,and was negatively correlated with corneal staining scoring and keratoconjunctival staining scoring(r s=0.270,P=0.025;r s=-0.331,P=0.006;r s=-0.325,P=0.007).MUC16 expression was positively correlated with TBUT,and was negatively correlated with blurred vision scoring,symptom exacerbation scoring during reading,impact scoring of driving at night,impact scoring of computer and impact scoring of TV use(r s=0.249,P=0.039;r s=-0.359,P=0.047;r s=-0.370,P=0.034;r s=-0.558,P=0.016;r s=-0.498,P=0.006;rs=-0.515,P=0.002).Conclusions The gene expressions of MUC1 and MUC16 are higher in conjunctiva of dry eye patients.MUC1 mRNA expression is related to patients'signs.MUC16 mRNA expression is related to the quality of life of patients.