摘要
目的:筛选并鉴定无排卵性功能失调性子宫出血(anovulatory dysfunctional uterine bleeding,ADUB)患者血清中的蛋白标志物,并检测被筛选蛋白标志物在ADUB患者月经血中的表达,探讨其与ADUB的相关性。方法:采集128例ADUB患者、93例月经正常妇女(对照组)的血液标本,分离血清,同时收集她们的月经血,分离上清。应用CM10蛋白芯片进行蛋白质检测,采用Biomarker WizardTM3.2软件分析检测结果。应用Tricine-SDS-PAGE胶分离、质谱鉴定和免疫共沉淀对检测到有差异的蛋白质峰进行蛋白质鉴定。然后采用ELISA,RT-PCR和Western印迹检测被筛选标志蛋白在ADUB患者月经血上清中的表达。结果:筛选到5个有差异的蛋白质峰,其中ADUB患者组在11.80km/z的蛋白质峰强度值明显大于对照组(P<0.05),在13.59km/z的蛋白质峰强度值明显低于对照组(P<0.05),两组在蛋白质峰为13.79,13.85和14.20km/z的强度值差异没有统计学意义(P>0.05)。11.80km/z的蛋白质峰被鉴定为血清淀粉样蛋白A(SAA),13.59km/z的蛋白质峰被鉴定为血管内皮细胞生长因子(VEGF)。在ADUB患者月经血上清中,SAA表达显著高于对照组(P<0.05),而VEGF的表达明显低于对照组(P<0.05)。结论:成功筛选和鉴定出与ADUB相关的两个血清蛋白标志物SAA和VEGF,ADUB患者SAA表达上调,而VEGF表达下调,这是ADUB病因或疾病发生结果还有待进一步证实。
Objective To screen and identify the serum biomarker of anovulatory dysfunctional uterine bleeding （ ADUB ） , to determine the expression of biomarker protein in menses of ADUB patients, and to investigate the relation between ADUB and the biomarker proteins. Methods Subjects included 128 ADUB patients and 93 age-matched controls （normal women）. Their serum and supernatant of mense were collected and stored for use at - 80℃. The differential proteins in the serum of the 2 groups were detected by CM 10 and analyzed by Biomarker WizardTM 3. 2 software. Then, the differential proteins were identified by Tricine-SDS-PAGE gel separation, spectrometry identifica-tion, and immunoprecipitation. The expression of the protein identified above in the menses was test- ed by ELISA, RT-PCR, and Western blotting. SPSS 14. 1 was applied for statistical analysis and chart drawing. Results Five differential protein peaks were screened and their peak values were 11.80, 13.59, 13.79, 13. 85, and 14. 20 km/z, respectively. The intensity of protein peak （ 11.80 km/z ） which was identified as serum amyploid protein A （ SAA ） of ADUB was significantly higher than that of the controls （P 〈 0. 05 ）. While the intensity of protein peak （ 13.59 km/z ） which was identified as vascular endothelial growth factor （ VEGF ） of ADUB was obviously lower than that of the controls （P 〈 0.05 ）. The intensity of protein peak 13.08, 13.85, and 14.20 was not different between the cases and controls. SAA expressed highly in the menses of ADUB but low in that of the controls. Conversely, VEGF expressed highly in the menses of the control but low in that of the ADUB. Conclusion Two biomarkers which might be related with ADUB have been correctly screened and identified as SAA and VEGF. It needs further study whether the increased expression of SAA and reduced expression of VEGF are the cause or result of ADUB.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2009年第7期616-623,共8页
Journal of Central South University ：Medical Science
基金
海南省自然科学基金(30633)~~
关键词
无排卵性功能失调性子宫出血
蛋白标志物
血清淀粉样蛋白A
血管内皮细胞生长因子
表面加强激光解析离子化飞行时间质谱
anovulatory dysfunctional uterine bleeding （ADUB）
biomarker
serum amyploid protein A （ SAA ）
vascular endothelial growth factor （ VEGF ）
surface-enhanced laser desorption ionization time-of-flight mass spectrometry （SELDI-TOF-MS）
