第一临床医学院知识库

目的：本文旨在通过研究速激肽human hemokinin-1碳端类似物hHK-1（4-11）和氮端类似物hHK-1（1-7）、hHK-1（1-8）、hHK-1（1-9）、hHK-1（1-10）对精子活力的影响，通过这种方法找出速激肽作用于精子活力的最有效片段，通过这样的研究希望能为提高精子活力的药物提供新的思路，为临床的体外受精-胚胎移植以及卵胞浆内单精子注射等辅助生殖治疗手段中保持精子活力的药物提供新的线索。

方法：通过化学合成法——多肽固相合成方法合成并验证了5个不同的人速激肽的片段，它们分别是人速激肽的碳端片段hHK-1（4-11），以及人速激肽的氮端类似物hHK-1（1-7）、hHK-1（1-8）、hHK-1（1-9）和hHK-1（1-10）。首先，我们分别检测了hHK-1（4-11）、hHK-1（1-7）、hHK-1（1-8）、hHK-1（1-9）和hHK-1（1-10）对正常精液PR级和PR+NP级精子活力影响，其次，我们通过流式细胞仪分析了正常成熟精子与上述药物共培养3小时和6小时后精子DNA碎片的产生情况。

结果：不同浓度（1×10-3mol/L、1×10-4mol/L、1×10-5mol/L）的hHK-1（4-11）、hHK-1（1-7）、hHK-1（1-8）、hHK-1（1-9）和hHK-1（1-10）分别作用于精子，hHK-1（1-7）作用于正常精子90min后的PR和PR+NP级精子百分率和空白对照組比起初始的PR和PR+NP级精子百分率，精子百分率的降低具有统计学意义（P<0.05），但是hHK-1（4-11）、hHK-1（1-8）、hHK-1（1-9）和hHK-1（1-10）作用于正常精子90min的PR和PR+NP级精子百分率的降低无统计学差异。

结论：hHK-1（4-11）、hHK-1（1-8）、hHK-1（1-9）和hHK-1（1-10）具有保持精子活力的作用，而hHK-1（1-7）并不具备保持精子活力的作用。另一方面，通过精子DNA碎片的分析结果显示：所有的速激肽类似物均没有使精子DNA碎片率上升，这就说明，这些药物并不存在精子毒性作用，其本身并不会对精子活力及完整性产生影响。

Objective: This paper aims to study the effect of C-terminal analogues of human hemokinin-1 [ hHK-1 (4-11) ]and the N-terminal analogues of human hemokinin-1 [hHK-1(1-7), hHK-1 (1-8), hHK-1 (1-9), hHK-1 (1-10)] on sperm motility, In this way, we aim to find out the most effective sequence on sperm motility. Through this study we hope to, on the one hand, provide a new way for drugs to improve sperm motility, on the other hand, to provide new clues for drug to maintain sperm in the process of assisted reproductive in vitro fertilization-embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI).

Methods: In the initial of this study, the members of the study successfully obtained five different human Hemokinin-1 (hHK-1) fragments by solid-phase peptide synthesis method, which were hHK-1 (4-11), the C-terminal of the human hemokinin-1, and the N-terminal of the human hemokinin-1, hHK-1 (1-7), hHK-1 (1-8), hHK-1 (1-9) and hHK-1 (1-10). In our study, we firstly measured the effects of hHK-1 (4-11), hHK-1 (1-7), hHK-1 (1-8), hHK-1 (1-9) and hHK-1 (1-10) on normal semen PR and PR+NP sperm motility, and secondly, we analyzed the production of sperm DNA fragments after incubated with the above drugs for 3 hours and 6 hours by flow cytometry.

Results: The results showed that the decrease of the rate of PR and PR+NP in group hHK-1(1-7) and blank control have significant difference (P<0.05) after incubated with different concentrations of hHK-1(4-11), hHK-1(1-7), hHK-1(1-8), hHK-1(1-9), hHK-1(1-10). However, the decrease of the rate of PR and PR+NP in group hHK-1(4-11), hHK-1(1-8), hHK-1(1-9), hHK-1(1-10) have no significant difference.

Conclusion: This illustrated that, on the one hand, hHK-1(4-11), hHK-1(1-8), hHK-1(1-9), hHK-1(1-10) play a role in maintaining sperm motility, however, hHK-1(1-7) do not have the effect of maintaining sperm motility. On the other hand, the result of DFI showed that all the tachykinin analogues have no impact on the sperm DNA fragmentation, this indicating that these drugs do not have toxicity on sperm, which in itself does not affect sperm motility and integrity.