Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Rahmatieh, Shahrekord, 8813833435, Iran.
Genetics Division, Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran.
Reprod Sci. 2021 Jul;28(7):2060-2069. doi: 10.1007/s43032-021-00485-7. Epub 2021 Feb 23.
Numerous evidences suggested that microRNAs (miRs) could play an active and significant role during spermatogenesis. Cysteine-rich secretory protein (CRISP3) has a role in inflammatory response and is extremely over-expressed in adolescents with varicocele seminal plasma and modified semen analysis. Nowadays, the miRs expression's association with their target genes is well recognized. The aim of this study was evaluating the association of CRISP3 and four candidate miRs among teratozoospermia (TZ) infertile men. First, we have selected four miRs, miR-182-5p, miR-192-5p, miR-204-5p, and miR-493-5p bioinformatically. After that, RNA was extracted from semen samples of 21 TZ patients and 20 normozoospermia (Norm). Then, their expression levels were assessed using real-time polymerase chain reaction method. In the next step, we quantified the expression of two CRISP3 protein isoforms, targeted by these miRs, using western blotting. According to our results, up-regulation of miR-182-5p, miR-192-5p, and miR-493-5p was observed. MiR-182-5p, miR-192-5p, and miR-493-5p showed good AUC values which can be introduced as possible biomarkers of TZ. In addition, the expression level of the CRISP3 glycosylated (31 kDa) isoform was significantly lower in TZ patients than Norm ones. Notably, in TZ patients, there was a possibly positive correlation of glycosylated CRISP3 expression with normal sperm morphology. According to our results, CRISP3 protein can play a significant role in male infertility especially in maturation formation of spermatozoa. Also, deregulation of the studied miRs, miR-182-5p, miR-92-5p, and miR-493-5p, can suggest a regulatory network between these miRs and CRISP3 isoforms and suggest their regulatory roles in male infertility.
大量证据表明,微小 RNA(miRs)在精子发生过程中可能发挥积极而重要的作用。富含半胱氨酸的分泌蛋白(CRISP3)在炎症反应中起作用,在精索静脉曲张精液和改良精液分析的青少年中表达水平极高。如今,miRs 表达与其靶基因的关系已得到充分认识。本研究旨在评估 CRISP3 与 4 种候选 miRs 在畸形精子症(TZ)不育男性中的关联。首先,我们通过生物信息学方法选择了 4 种 miR,miR-182-5p、miR-192-5p、miR-204-5p 和 miR-493-5p。然后,从 21 例 TZ 患者和 20 例正常精子症(Norm)患者的精液样本中提取 RNA,然后使用实时聚合酶链反应方法评估其表达水平。下一步,我们使用 Western blot 定量这些 miR 靶向的两种 CRISP3 蛋白异构体的表达。根据我们的结果,观察到 miR-182-5p、miR-192-5p 和 miR-493-5p 的上调。miR-182-5p、miR-192-5p 和 miR-493-5p 表现出良好的 AUC 值,可以作为 TZ 的可能生物标志物。此外,与 Norm 相比,TZ 患者的 CRISP3 糖基化(31 kDa)异构体的表达水平显着降低。值得注意的是,在 TZ 患者中,糖基化 CRISP3 表达与正常精子形态之间可能存在正相关关系。根据我们的结果,CRISP3 蛋白在男性不育症中可能发挥重要作用,特别是在精子发生的成熟形成中。此外,研究的 miRs,miR-182-5p、miR-92-5p 和 miR-493-5p 的失调,可以提示这些 miRs 和 CRISP3 异构体之间的调控网络,并提示它们在男性不育症中的调控作用。