本文检查了胱抑素S(CST4)的泪液浓度,钙周期素(S100A6),钙粒蛋白A(S100A8),和基质金属蛋白酶9(MMP9),以及生物标志物表达之间的相关性,临床参数,和患有干眼症(DE)的患者的疾病严重程度。通过ELISA测试和用于蛋白质定量的定制抗体微阵列获得结果的比较。
这个单中心,观察性研究招募了59名参与者(45名DE和14名对照).临床评估包括眼表疾病指数(OSDI)问卷,泪液渗透压(OSM)测试,Schirmer试验(SCH),撕裂破裂时间(TBUT),荧光素(FLUO)和赖斯胺绿(LG)角膜染色,和睑板腺评估(MGE)。使用标准的单独ELISA测定法测量CST4、S100A6、S100A8和MMP9的撕裂浓度。还使用定制的多重抗体微阵列测量CST4、S100A6和MMP9的水平。评估变量之间的相关性,显著性水平为p值<0.05。
用ELISA定量泪液蛋白生物标志物表明,与对照(CT)受试者相比,DE患者泪液中CST4的浓度显着降低(2.14倍)(p<0.001)。与CT相比,DE患者泪液中的S100A6和S100A8浓度明显更高(分别为1.36和2.29倍;p<0.001和0.025)。DE患者的MMP9水平也较高(5.83倍),但不显著(p=0.22)。CST4和S100A6浓度的变化与干眼病(DED)严重程度显着相关。使用抗体微阵列对CST4、S100A6和MMP9进行定量,证实了ELISA结果。观察到类似的趋势:CST4减少1.83倍(p值0.01),S100A6增加8.63倍(p值<0.001),MMP9增加9.67倍(p值0.94),但灵敏度更高。生物标志物浓度与DED相关的体征和症状显著相关。
S100A6、S100A8和CST4诊断生物标志物与DED临床参数强烈相关。S100A6和CST4也可用于对DE严重程度进行分级。多重抗体微阵列技术,这里用于泪液多标记物定量,似乎比标准ELISA测试更敏感。
This paper examines the tear concentration of cystatin S (CST4), calcyclin (S100A6), calgranulin A (S100A8), and matrix metalloproteinase 9 (MMP9), and the correlation between biomarker expression, clinical parameters, and disease severity in patients suffering from dry eye (DE). A comparison of the results is obtained via ELISA tests and customized antibody microarrays for protein quantification.
This single-center, observational
study recruited 59 participants (45 DE and 14 controls). Clinical evaluation included an Ocular Surface Disease Index (OSDI) questionnaire, a tear osmolarity (OSM) test, the Schirmer test (SCH), tear breakup time (TBUT), fluorescein (FLUO) and lissamine green (LG) corneal staining, and meibomian gland evaluation (MGE). Tear concentrations of CST4, S100A6, S100A8, and MMP9 were measured using standard individual ELISA assays. The levels of CST4, S100A6, and MMP9 were also measured using customized multiplexed antibody microarrays. Correlations between variables were evaluated, and a significance level was p value <0.05.
The quantification of tear protein biomarkers with ELISA showed that the concentration of CST4 was significantly (2.14-fold) reduced in tears of DE patients in comparison with control (CT) subjects (p < 0.001). S100A6 and S100A8 concentrations were significantly higher in the tears of DE patients (1.36- and 2.29-fold; p < 0.001 and 0.025, respectively) in comparison with CT. The MMP9 level was also higher in DE patients (5.83-fold), but not significantly (p = 0.22). The changes in CST4 and S100A6 concentrations were significantly correlated with dry eye disease (DED) severity. Quantification of CST4, S100A6, and MMP9, using antibody microarrays, confirmed the ELISA results. Similar trends were observed: 1.83-fold reduction for CST4 (p value 0.01), 8.63-fold increase for S100A6 (p value <0.001) and 9.67-fold increase for MMP9 (p value 0.94), but with higher sensitivity. The biomarker concentrations were significantly associated with the signs and symptoms related with DED.
S100A6, S100A8, and CST4 diagnostic biomarkers strongly correlate with DED clinical parameters. S100A6 and CST4 are also useful for grading DE severity. The multiplexed antibody microarray technique, used here for tear multi-marker quantification, appears more sensitive than standard ELISA tests.