Department of Surgery, Division of Urology, Human Reproduction Section, Sao Paulo Federal University, Sao Paulo, Brazil.
Fertil Steril. 2013 Sep;100(3):667-72. doi: 10.1016/j.fertnstert.2013.04.036. Epub 2013 May 23.
To compare seminal plasma protein profiles before and after varicocele correction to assess if surgical intervention alters the protein profile.
Prospective study.
Academic research environment.
PATIENT(S): Nineteen adolescent boys with varicocele grades II or III.
INTERVENTION(S): Two semen samples were collected before bilateral subinguinal microsurgical varicocelectomy, and two semen samples were collected 3 months after surgery. Seminal plasma protein profiles were determined with the use of two-dimensional gel electrophoresis. Proteins were separated in 18-cm 3-10 pH strips and 10%-17.5% gradient gels. Gels were stained, scanned, and compared with the use of Imagemaster 2D platinum 7.0. Spots of interest were removed from gels, and protein digestion was performed with the use of trypsin. Digests were identified with the use of electrospray ionization-quadrupole/time-of-flight tandem mass spectrometry (ESI-QTOF MS/MS), and spectra were analyzed with the use of the Mascot software.
MAIN OUTCOME MEASURE(S): Proteins uniquely or overexpressed in each period (before or after varicocelectomy).
RESULT(S): Nineteen spots were differentially expressed between pre- and postsurgery samples. Identified proteins were albumin, proteasome subunit alpha type 6, alpha-1-antitrypsin, fibronectin, CD177, prostatic acid phosphatase, specific prostatic antigen, alpha-2-antiplasmin, vitamin D-binding protein, gastricsin, clusterin, semenogelin-1, semenogelin-2, superoxide dismutase, protein-glutamine gamma glutamyltransferase-4, and prolactin-inducing protein.
CONCLUSION(S): Varicocelectomy is associated with changes in the seminal plasma protein profile. Understanding specific pathways leading to male infertility may further assist physicians in demonstrating deviation from homeostasis in male infertility. In addition, it may be possible to observe if surgical intervention does indeed revert altered pathways toward a homeostatic state.
比较精索静脉曲张矫正前后的精浆蛋白图谱,以评估手术干预是否改变蛋白图谱。
前瞻性研究。
学术研究环境。
19 名精索静脉曲张 II 级或 III 级的青少年男性。
双侧精索下显微精索静脉曲张结扎术前采集 2 份精液样本,术后 3 个月采集 2 份精液样本。使用二维凝胶电泳法测定精浆蛋白图谱。使用 18-cm 3-10 pH 条和 10%-17.5%梯度凝胶分离蛋白质。凝胶染色、扫描,并用 Imagemaster 2D platinum 7.0 进行比较。从凝胶中取出感兴趣的斑点,用胰蛋白酶进行蛋白消化。消化物用电喷雾电离-四极杆/飞行时间串联质谱(ESI-QTOF MS/MS)鉴定,用 Mascot 软件分析谱图。
每个时期(手术前后)特有的或过表达的蛋白质。
手术前后样本中差异表达的有 19 个斑点。鉴定出的蛋白质有白蛋白、蛋白酶体亚基α型 6、α-1 抗胰蛋白酶、纤维连接蛋白、CD177、前列腺酸性磷酸酶、特异性前列腺抗原、α-2-抗纤溶酶、维生素 D 结合蛋白、胃蛋白酶、聚集蛋白、精液蛋白 1、精液蛋白 2、超氧化物歧化酶、蛋白谷氨酰胺γ-谷氨酰转移酶 4 和催乳素诱导蛋白。
精索静脉曲张结扎术与精浆蛋白图谱的变化有关。了解导致男性不育的特定途径可能进一步帮助医生发现男性不育症中偏离内稳态的情况。此外,还可以观察手术干预是否确实使改变的途径恢复到内稳态。