目的 研究女性雄激素性脱发(FAGA)患者血浆中代谢相关蛋白及其价值。 方法 2021年3月至2023年3月从复旦大学附属华山医院皮肤科门诊招募年龄18～50岁的FAGA患者(FAGA组)及健康对照女性(HC组)。采集每位受试者的外周静脉血3 ml，离心获得血浆。对血浆行Olink蛋白质组学分析，通过R语言筛选2组间的差异表达蛋白，并以受试者工作特征(ROC)曲线评估差异蛋白的诊断性能；对差异表达蛋白行基因本体论(GO)分析。取FAGA组顶区、HC组枕区毛囊行免疫荧光分析，对筛选的差异表达蛋白进行验证。应用SPSS 25.0软件对数据进行分析，正态分布计量资料以±s表示，2组受试者基本资料比较、毛囊中差异蛋白相对荧光强度比较采用t检验；对血浆中代谢相关蛋白与受试者基本资料行Pearson相关分析。P<0.05为差异有统计学意义。 结果 FAGA组纳入61例患者，年龄(33.8±7.4)岁，发病年龄(29.5±7.8)岁，其中轻度38例、中度14例、重度9例；HC组纳入27例健康女性，年龄(32.0±7.7)岁。2组受试者的基本资料(年龄、体重指数及睾酮、25-羟维生素D、尿酸、铁蛋白水平)比较，差异均无统计学意义(P>0.05)。与HC组相比，FAGA组血浆中有26种差异蛋白表达显著上调(P<0.05)，其中AHCY、NECTIN2差异最为显著(P均＝0.003)，ROC曲线评估显示AHCY和NECTIN2的曲线下面积均大于0.7，表现出良好的诊断准确性。GO分析显示，差异蛋白主要富集在BAT3复合物(细胞组分)，泛素依赖性ERAD途径、自然杀伤细胞活化(生物学过程)，以及泛素蛋白连接酶结合、泛素特异性蛋白酶结合(分子功能)等方面。Pearson相关分析显示，AHCY(r＝－0.23，P＝0.010)、NECTIN2(r＝－0.31，P＝0.033)均与FAGA患者的脱发严重程度呈负相关。毛囊免疫荧光分析结果显示，FAGA组中AHCY、NECTIN2相对荧光强度高于HC组(P<0.05)，即AHCY、NECTIN2均在FAGA组中表达上调。 结论 代谢相关蛋白在FAGA中起着重要作用，AHCY及NECTIN2有可能作为FAGA的早期诊断生物标志物。 Objective To investigate the metabolism-related proteins and their presence in the plasma of female androgenetic alopecia (FAGA) patients. Methods From March 2021 to March 2023, FAGA patients aged 18-50 (FAGA group) and healthy women (HC group) were recruited from the Dermatology Outpatient Department of Huashan Hospital. 3 ml of peripheral venous blood was collected from each participant and centrifuged to obtain plasma. Olink proteomics analysis was performed on the collected plasma, differentially expressed proteins were screened with R language, the diagnostic accuracy of the differentially expressed proteins was assessed using receiver operating characteristic (ROC) curve. Gene ontology (GO) analysis was performed on differentially expressed proteins. Immunofluorescence analysis on hair follicles in the parietal region of the FAGA group and the occipital region of the HC group was performed to validate the differentially expressed proteins identified. SPSS 25.0 software was used to analyze the data, with normal distribution metric data represented by Mean±SD. Student’s t-test was used to compare the basic information of two groups of subjects and the relative fluorescence intensity of differentially expressed proteins in hair follicles. Pearson correlation analysis was performed on plasma metabolism-related proteins and the basic information of subjects. P<0.05 indicates a statistically significant difference. Results Sixty-one cases were included in the FAGA group, with an average age of (33.8±7.4) years and an onset age of (29.5±7.8) years. Among them, 38 cases were mild FAGA, 14 cases were moderate, and 9 cases were severe. Twenty-seven cases were included in the HC group, with an average age of (32.0±7.7) years. There was no statistically significant difference in the basic information (age, body mass index, testosterone, 25-hydroxyvitamin D, uric acid, and ferritin levels) between the two groups of subjects (P>0.05). Compared to the HC group, the plasma of the FAGA group showed 26 significantly upregulated differentially expressed proteins (P<0.05), with AHCY and NECTIN2 exhibiting the most significant differences (allP=0.003). The ROC curve evaluation revealed that the area under the curve for AHCY and NECTIN2 was greater than 0.7, indicating good diagnostic accuracy. The GO analysis revealed that the differentially expressed proteins were primarily enriched in the BAT3 complex (cellular component), ubiquitin-dependent ERAD pathway, natural killer cell activation (biological process), as well as ubiquitin protein ligase binding and ubiquitin-specific protease binding (molecular function). Pearson correlation analysis revealed that AHCY (r=-0.23, P=0.010) and NECTIN2 (r=-0.31, P=0.033) were negatively correlated with the severity of hair loss in FAGA patients. The results of hair follicle immunofluorescence analysis showed that the relative fluorescence intensity of AHCY and NECTIN2 in the FAGA group was higher than that in the HC group (P<0.05). In other words, both AHCY and NECTIN2 were upregulated in the FAGA group. Conclusion Metabolism-related proteins play an important role in FAGA. AHCY and NECTIN2 may serve as early diagnostic biomarkers for FAGA.
A proteomics research on metabolism-related proteins in female androgenetic alopecia
Objective To investigate the metabolism-related proteins and their presence in the plasma of female androgenetic alopecia (FAGA) patients. Methods From March 2021 to March 2023, FAGA patients aged 18-50 (FAGA group) and healthy women (HC group) were recruited from the Dermatology Outpatient Department of Huashan Hospital. 3 ml of peripheral venous blood was collected from each participant and centrifuged to obtain plasma. Olink proteomics analysis was performed on the collected plasma, differentially expressed proteins were screened with R language, the diagnostic accuracy of the differentially expressed proteins was assessed using receiver operating characteristic (ROC) curve. Gene ontology (GO) analysis was performed on differentially expressed proteins. Immunofluorescence analysis on hair follicles in the parietal region of the FAGA group and the occipital region of the HC group was performed to validate the differentially expressed proteins identified. SPSS 25.0 software was used to analyze the data, with normal distribution metric data represented by Mean±SD. Student’s t-test was used to compare the basic information of two groups of subjects and the relative fluorescence intensity of differentially expressed proteins in hair follicles. Pearson correlation analysis was performed on plasma metabolism-related proteins and the basic information of subjects. P<0.05 indicates a statistically significant difference. Results Sixty-one cases were included in the FAGA group, with an average age of (33.8±7.4) years and an onset age of (29.5±7.8) years. Among them, 38 cases were mild FAGA, 14 cases were moderate, and 9 cases were severe. Twenty-seven cases were included in the HC group, with an average age of (32.0±7.7) years. There was no statistically significant difference in the basic information (age, body mass index, testosterone, 25-hydroxyvitamin D, uric acid, and ferritin levels) between the two groups of subjects (P>0.05). Compared to the HC group, the plasma of the FAGA group showed 26 significantly upregulated differentially expressed proteins (P<0.05), with AHCY and NECTIN2 exhibiting the most significant differences (allP=0.003). The ROC curve evaluation revealed that the area under the curve for AHCY and NECTIN2 was greater than 0.7, indicating good diagnostic accuracy. The GO analysis revealed that the differentially expressed proteins were primarily enriched in the BAT3 complex (cellular component), ubiquitin-dependent ERAD pathway, natural killer cell activation (biological process), as well as ubiquitin protein ligase binding and ubiquitin-specific protease binding (molecular function). Pearson correlation analysis revealed that AHCY (r=-0.23, P=0.010) and NECTIN2 (r=-0.31, P=0.033) were negatively correlated with the severity of hair loss in FAGA patients. The results of hair follicle immunofluorescence analysis showed that the relative fluorescence intensity of AHCY and NECTIN2 in the FAGA group was higher than that in the HC group (P<0.05). In other words, both AHCY and NECTIN2 were upregulated in the FAGA group. Conclusion Metabolism-related proteins play an important role in FAGA. AHCY and NECTIN2 may serve as early diagnostic biomarkers for FAGA.
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