半乳糖凝集素10在不同类型鼻息肉中的表达及其发病机制中的作用

Expression and pathological role of galectin-10 in different types of nasal polyps

目的检测半乳糖凝集素10(Galectin-10)在不同程度嗜酸粒细胞浸润的鼻息肉中的表达差异,探讨Galectin-10是否可作为嗜酸粒细胞性鼻息肉(eosinophilic chronic rhinosinusitis with nasal polyps,ECRSwNP)的新型标志物,及其在ECRSwNP发病机制中的作用。方法纳入2018年11月至2019年4月在南昌大学第二附属医院住院行鼻内镜手术的患者共36例进行回顾性分析,其中男20例,女16例;年龄为14~74岁。36例患者分为3组:ECRSwNP组11例;非嗜酸粒细胞性鼻息肉(noneosinophilic chronic rhinosinusitis with nasal polyps,non-ECRSwNP)组15例;对照组单纯鼻中隔偏曲患者10例。采用HE染色对CRSwNP进行组织学评估并将其分为ECRSwNP和non-ECRSwNP。根据患者有无变应性鼻炎,将CRSwNP患者分为AR组及非AR组(non-AR),将CRSwNP与对照组根据是否属于特应性状态分为特应性状态阳性组(Atopy)及阴性组(non-Atopy)。免疫组织化学测定Galectin-10在ECRSwNP、non-ECRSwNP及对照组组织中的阳性定位及半定量表达、蛋白免疫印迹(Western Blot)检测Galectin-10蛋白分别在3组中的表达水平、实时荧光定量反转录聚合酶链反应检测Galectin-10 mRNA在3组中的不同表达;分析Galectin-10的表达与临床因素,如合并变应性鼻炎(allergic rhinitis,AR)、特应性状态是否相关。运用IBM SPSS 19.0及Graphpad prism 7.0对实验数据进行处理与作图。结果免疫组织化学染色结果提示Galectin-10主要位于嗜酸粒细胞,与non-ECRSwNP组(0.028±0.004)和对照组(0.025±0.004)相比,ECRSwNP组中Galectin-10的半定量表达(0.051±0.003)更高,差异有统计学意义(t值分别为3.862、5.137,P值均<0.01),non-ECRSwNP组与对照组相比,差异无统计学意义(t=0.560,P>0.05)。Western Blot法检测Galectin-10蛋白的表达发现,与non-ECRSwNP组和对照组相比,ECRSwNP组中Galectin-10蛋白的表达更高,差异有统计学意义(t值分别为25.351、27.376,P值均<0.01),non-ECRSwNP组与对照组相比,差异无统计学意义(t=1.071,P>0.05)。Galectin-10 mRNA在3组中的表达:与non-ECRSwNP组(1.188±0.054)和对照组(1.020±0.142)相比,ECRSwNP组Galectin-10 mRNA表达量(2.413±0.303)较高,差异有统计学意义(t值分别为3.973、4.156,P值均<0.05),non-ECRSwNP组与对照组相比,差异无统计学意义(t=1.110,P>0.05)。采用免疫组织化学方法检测组间Galectin-10蛋白的半定量表达差异,结果显示AR组与non-AR组比较,Galectin-10的表达差异无统计学意义(P值均>0.05),特应性状态阳性组(Atopy)与特应性状态阴性组(non-Atopy)相比,Galectin-10的表达差异无统计学意义(P值均>0.05)。结论Galectin-10在ECRSwNP中表达升高,但是在AR与non-AR以及在特应性状态阳性组(Atopy)与特应性状态阴性组(non-Atopy)患者间差异无统计学意义,提示Galectin-10可作为ECRSwNP的新型生物标志物,并可能在非变态反应诱发的ECRSwNP发病机制中具有意义。

Objective:To investigate the different expression of galectin-10 in nasal polyps with different degrees of eosinophil infiltration,and to explore whether galectin-10 can be used as a new biomarker of eosinophilic chronic rhinosinusitis with nasal polyps(ECRSwNP)and its possible role in the pathogenesis of ECRSwNP.Methods:A total of 36 patients(20 males,16 females,aged from 14 to 74 years old)who underwent functional endoscopic sinus surgery at the Second Affiliated Hospital of Nanchang University from November 2018 to April 2019 were enrolled into the retrospective study,including 11 cases of ECRSwNP,15 cases of non-ECRSwNP and 10 cases in control group(deviation of nasal septum).The patients were divided into allergic rhinitis and non-allergic rhinitis groups,atopy and non-atopy groups according to whether patients in the experimental group and control group had allergic rhinitis and atopy.HE staining was performed for histological assessment of CRSwNP which was classfied as ECRSwNP and non-ECRSwNP.Immunohistochemistry(IHC)was used to determine the positive localization and semi-quantitative expression level of galectin-10 protein in ERSwNP,non-ECRSwNP and control groups.The expression levels of galectin-10 protein in three groups were determined by Western Blot.The expression levels of galectin-10 mRNA in three groups were determined using real-time quantitative polymerase chain reaction(qRT-PCR).Analyzing the correlation between the expression of galectin-10 and clinical factors including the allergic rhinitis and atopy,SPSS 19.0 software and Graphpad prism 7.0 were used for statistical analysis and mapping.Results:By using IHC method,it was found that galectin-10 was mainly localized in eosinophils in the polyp tissues.The semi-quantitative expression of the galectin-10 in the ECRSwNP group(0.051±0.003)was significantly higher than that of non-ECRSwNP(0.028±0.004)and control groups(0.025±0.004,t value was 3.862 and 5.137,both P<0.01).There was no significant difference between the control and non-ECRSwNP groups(t=0.560,P>0.05).The expression of galectin-10 in the ECRSwNP group was significantly higher than that of non-ECRSwNP and control groups(t value was 25.351 and 27.376,both P<0.01).However,there was no significant difference between the non-ECRSwNP and control groups(t=1.071,P>0.05).Compared with the non-ECRSwNP(1.188±0.054)and control groups(1.020±0.142),the expression of galectin-10 mRNA was higher in the ECRSwNP group(2.413±0.303),the differences were significant(t value was 3.973 and 4.156,both P<0.05).There was no significant difference between the non-ECRSwNP and control groups(t=1.110,P>0.05).There was no significant difference in the expression of galectin-10 between the allergic rhinitis group and the non-allergic rhinitis group(all P>0.05),so as to the atopy group and non-atopy group(all P>0.05).Conclusion:The expression level of galectin-10 is elevated in ECRSwNP,and not influenced by allergic status,suggesting that galectin-10 may be a new biomarker for ECRSwNP and play an important role in the pathogenesis of ECRSwNP.