脑胶质瘤组织和患者脑脊液中淀粉样前体蛋白水平与胶质瘤病理等级间的关系

Relations of glioma pathological grades with amyloid precursor protein content in brain glioma tissues and cerebrospinal fluid of these patients

目的探讨淀粉样前体蛋白(APP)在脑胶质瘤组织和患者脑脊液中的表达水平及意义。方法(1)收集西南医科大学附属中医医院神经外科自2015年1月至2020年12月手术切除并经病理确诊的脑胶质瘤标本56例,其中世界卫生组织(WHO)分级Ⅰ级9例、Ⅱ级14例、Ⅲ级18例、Ⅳ级15例,另外收集同期4例高血压脑出血患者行血肿清除术时切除的坏死脑组织作为对照,采用Western blotting、实时荧光定量PCR(RT-qPCR)分别检测脑胶质瘤组织和对照脑组织APP蛋白和mRNA的表达。(2)除了上述56例脑胶质瘤患者,另外选择同期收治的4例病毒性脑膜炎、4例细菌性脑膜炎、4例自身免疫性脑炎、5例癫痫和3例偏头痛患者及3例正常对照者,采用ELISA法检测所有受试者脑脊液中APP含量。结果(1)Western blotting、RT-qPCR检测结果显示WHO分级Ⅰ、Ⅱ、Ⅲ、Ⅳ级胶质瘤组织APP蛋白和mRNA的表达均高于对照脑组织,且WHO分级Ⅰ级和Ⅱ级、Ⅲ、Ⅳ级脑胶质瘤组织APP蛋白和mRNA的表达依次增高,差异均有统计学意义(P<0.05)。WHO分级Ⅰ级和Ⅱ级脑胶质瘤中,首发组与复发组间脑胶质瘤APP蛋白和mRNA表达的差异均无统计学意义(P>0.05);WHO分级Ⅲ级和Ⅳ级脑胶质瘤中,复发组脑胶质瘤APP mRNA的表达均高于首发组,差异均有统计学意义(P<0.05)。(2)ELISA检测结果显示,病毒性脑膜炎、细菌性脑膜炎、自身免疫性脑炎、癫痫和偏头痛患者与正常对照者脑脊液中APP含量的差异无统计学意义(P>0.05)。WHO分级Ⅰ、Ⅱ、Ⅲ、Ⅳ级脑胶质瘤患者脑脊液中APP含量均高于正常对照者,且WHO分级Ⅰ级和Ⅱ级、Ⅲ、Ⅳ级脑胶质瘤患者脑脊液中APP含量依次增高,差异均有统计学意义(P<0.05)。结论脑胶质瘤组织和胶质瘤患者脑脊液中APP呈高表达,其含量与脑胶质瘤病理等级有关,脑脊液中APP表达量可作为早期筛查脑胶质瘤患者的新方法。

Objective To investigate the amyloid precursor protein(APP)expression in brain glioma tissues and cerebrospinal fluid of glioma patients and their significances.Methods(1)Fifty-six brain glioma specimens surgically resected and pathologically confirmed in our hospital from January 2015 to December 2020 were collected,including 9 with WHO grading I,14 with WHO grading II,18 with WHO grading III,and 15 with WHO grading IV.In addition,the necrotic brain tissues of 4 patients with hypertensive intracerebral hemorrhage during hematoma removal at the same time period were collected as controls.Western blotting and real-time quantitative PCR(RT-qPCR)were used to detect the APP protein and mRNA expressions in glioma tissues and control brain tissues,respectively.(2)The cerebrospinal fluid of above 56 patients with glioma,4 patients with viral meningitis,4 patients with bacterial meningitis,4 patients with autoimmune encephalitis,5 patients with epilepsy,3 patients with migraine and 3 normal subjects were collected at the same time period,and the APP content in the cerebrospinal fluid of all subjects were detected by ELISA.Results(1)Western blotting and RT-qPCR showed that the APP protein and mRNA expressions in glioma tissues of WHO grading I,II,III and IV were significantly higher than those in control brain tissues,and the APP protein and mRNA expressions in glioma tissues of WHO grading I,II,III and IV increased successively,with statistical differences(P<0.05).There were no significant differences in APP protein and mRNA expressions between first-onset glioma and recurrent glioma in glioma tissues of WHO grading I and II(P>0.05);while in glioma tissues of WHO grading III and IV,APP mRNA expression in recurrent glioma was statistically higher than that in first-onset glioma(P<0.05).(2)ELISA results showed that APP content in patients with viral meningitis,bacterial meningitis,autoimmune encephalitis,epilepsy or migraine was not significantly different as compared with that in normal controls(P>0.05).The APP content in cerebrospinal fluid of patients with WHO grading I,II,III and IV gliomas was significantly higher than that in normal controls,and the APP content in cerebrospinal fluid of patients with WHO grading I,II,III and IV gliomas increased successively,with statistical differences(P<0.05).Conclusion APP is highly expressed in glioma tissues and cerebrospinal fluid of patients with glioma,and its expression is significantly correlated with the pathological grades of glioma;the detection of APP expression in cerebrospinal fluid can be used as a new screening method for glioma patients.