摘要
目的:研究B淋巴细胞瘤-2基因(Bcl-2)、 Bcl-2相关X蛋白(Bax)、细胞紧密连接蛋白(CLAUDIN)-11在不同严重程度生精功能障碍患者睾丸组织中的表达,探讨其临床意义。方法:收集80例睾丸组织标本,分为生精功能正常组、轻度生精功能障碍组、重度生精功能障碍组及唯支持细胞综合征组各20例,采用免疫组化法、实时荧光定量PCR(qRT-PCR)检测睾丸组织中Bcl-2、Bax及CLAUDIN-11的表达情况;采用电化学发光免疫分析法检测患者血清中卵泡刺激素(FSH)、催乳素(PRL)、黄体生成素(LH)、睾丸酮(T)水平。结果:免疫组化结果表明,Bcl-2及Bax主要表达于生精细胞的细胞质中,CLAUDIN-11在生精小管中有较高的阳性表达;qRT-PCR结果表明,与生精功能正常组相比,唯支持细胞综合征组患者的睾丸组织中Bax及CLAUDIN-11 mRNA表达水平显著增加(P﹤0.05),而Bcl-2 mRNA表达水平显著降低(P﹤0.05);与生精功能正常组患者相比,重度生精功能障碍组和唯支持细胞综合征组患者血清中FSH、PRL、 LH水平增高,T水平降低,差异有统计学意义(P<0.05);流式细胞分析表明,在轻度生精功能障碍组、重度生精功能障碍组患者中细胞凋亡率显著高于生精功能正常组患者,差异有统计学意义(P<0.05)。结论:睾丸组织中Bax蛋白异常上调,Bcl-2蛋白表达下调,加速了生精细胞的凋亡;同时CLAUDIN-11在睾丸组织中的异常上调,致使生精上皮空间构象发生异常,在患者生精功能障碍的发生发展中发挥着重要作用。
Objective: To study the expression of B-cell lymphoma-2(Bcl-2), Bcl-2 associated X protein(Bax), cellular tight junction protein(CLAUDIN)-11 in testicular tissue of patients with different types of spermatogenic dysfunction and to explore its clinical significance. Methods: 80 specimens of testicular tissue were collected, which were divided into normal spermatogenic function group, mild spermatogenesis dysfunction group, severe spermatogenesis dysfunction group and sertoli cell only syndrome group. The expressions of Bcl-2, Bax and CLAUDIN-11 in different testis tissues were detected by immunohistochemistry and qRT-PCR. The levels of follicle-stimulating hormone(FSH),prolactin(PRL), luteinizing hormone(LH), testosterone(T) in serum were detected by electrochemiluminescence immunoassay. Results: Immunohistochemical results showed that Bcl-2 and Bax were mainly expressed in the cytoplasm of spermatogenic cells, while CLAUDIN-11 was more expressed in the seminiferous tubules. The results of qRT-PCR showed that the expression levels of Bax and CLAUDIN-11 in testicular tissues of sertoli cell only syndrome group were significantly higher than those of normal spermatogenic function group(P<0.05), while the expression of Bcl-2 was significantly lower(P<0.05). Compared with the normal spermatogenic function group, the serum levels of FSH, PRL and LH increased and T decreased in severe spermatogenesis dysfunction group and sertoli cell only syndrome group(P<0.05). Flow cytometry analysis showed that apoptotic rate was significantly higher in mild spermatogenic dysfunction group and severe spermatogenic dysfunction group than in normal spermatogenic function group(P<0.05). Conclusion: Abnormal up-regulation of Bax and down-regulation of Bcl-2 protein in testicular tissue accelerate spermatogenic cell apoptosis. At the same time, the abnormal up-regulation of CLAUDIN-11 in testicular tissue results in abnormal spatial conformation of spermatogenic epithelium, which plays an important role in the occurrence and development of spermatogenic dysfunction.
作者
姚佳沛
徐建春
刘炜
樊凡
YAO Jiapei;XU Jianchun;LIU Wei;FAN Fan(The Second Department of Urology,the First Affiliated Hospital of Kunming Medical University,Kunming 650031,China)
出处
《东南大学学报(医学版)》
CAS
2019年第5期830-836,共7页
Journal of Southeast University(Medical Science Edition)
基金
2018年度云南省科技厅-昆明医科大学应用基础研究联合基金资助项目[2018FE001(-2220)]