Sun Y-L, Ping Z-G, Li C-J, Sun Y-F, Yi K-L, Chen L, Li X-Y, Wang X-L, Zhou X
College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, Jilin Province, China.
Reprod Domest Anim. 2011 Oct;46(5):889-95. doi: 10.1111/j.1439-0531.2011.01760.x. Epub 2011 Mar 3.
Follicular fluid (FF) includes various biologically active proteins which can affect follicular growth and maturation. Certain proteins could reflect the physiological and pathological status of follicles. The aim of the present study was to explore the key proteins associated with pathogenesis of follicular cysts, some of which may be candidate biomarkers for the condition. We analysed the proteomes of FF from small, medium, large and cystic follicles by two-dimensional electrophoresis (2-DE) combined with mass spectrometry (MS). The protein components in FF were found to be significantly different among groups; about 300 proteins spots in each group were examined, and 32 differentially expressed proteins were identified from different groups. To further reveal the source of identified proteins, transcripts encoding two of these, transferrin and RBP-4, were detected in granulosa cells (GCs) by RT-PCR, as well as the proteins were detected in 24 h culture media of GCs by ELISA. High levels of RBP-4 were examined in FF of cystic follicles by 2-DE analysis, which were significantly different to those in large follicles (p < 0.05). In conclusion, the study enriches our understanding of the proteins of FF; RBP-4 and transferrin originate from passive transfer and follicular synthesized secretion, and RBP-4 might be a candidate biomarker for porcine follicular cysts. Combined with histological studies, these results further suggest that changes of the type and quantity of proteins in FF might be attributed to an abnormal metabolism of follicular cells and structure of follicular wall in cystic follicles. Our findings will contribute to further insight into the pathogenesis of follicular cysts.
卵泡液(FF)包含多种生物活性蛋白,这些蛋白可影响卵泡的生长和成熟。某些蛋白能够反映卵泡的生理和病理状态。本研究的目的是探索与卵泡囊肿发病机制相关的关键蛋白,其中一些可能是该病症的候选生物标志物。我们通过二维电泳(2-DE)结合质谱(MS)分析了来自小卵泡、中卵泡、大卵泡和囊肿卵泡的卵泡液蛋白质组。结果发现,卵泡液中的蛋白质成分在各组之间存在显著差异;每组检测约300个蛋白斑点,从不同组中鉴定出32种差异表达蛋白。为进一步揭示所鉴定蛋白的来源,通过RT-PCR在颗粒细胞(GCs)中检测了其中两种蛋白(转铁蛋白和RBP-4)的编码转录本,并且通过ELISA在颗粒细胞的24小时培养基中检测到了这些蛋白。通过二维电泳分析,在囊肿卵泡的卵泡液中检测到高水平的RBP-4,其与大卵泡中的水平有显著差异(p < 0.05)。总之,本研究丰富了我们对卵泡液蛋白质的认识;RBP-4和转铁蛋白分别源于被动转运和卵泡合成分泌,并且RBP-4可能是猪卵泡囊肿的候选生物标志物。结合组织学研究,这些结果进一步表明,卵泡液中蛋白质类型和数量的变化可能归因于囊肿卵泡中卵泡细胞代谢异常和卵泡壁结构异常。我们的研究结果将有助于进一步深入了解卵泡囊肿的发病机制。