目的：本篇研究是要鑑定精蟲DNA碎片化，粒線體功能缺陷與染色體倍數異常之發生率和在臨床上精液分析結果與受精率之間做相關性比較，另外一個研究是評估精蟲DNA碎片化與體外授精生殖執行能力之間的關聯性。 材料與方法：挑選10個精液品質極差（精蟲數量稀少，活動力低下及形態異常）OTA病患利用ICSI療程來分析，另外再找出10個精液正常經由IVF療程的病患做為對照組。精液利用Percoll不連續兩層（80%-50%）的密度梯度離心來製備，利用IVOS的精液分析儀來分析，DNA碎片化則是採用末端去氧核苷酸轉移酶介導的dUTP缺口末端標記法（TUNEL）分析。粒線體功能完整性則用ApoalertTM探針（JC-1）檢測精蟲粒線體膜電位的改變，而染色體倍數異常則以螢光原位雜交法來判定。此外又收集303個包括與不包括ICSI的IVF療程，偵測其精蟲DNA碎片化的百分比與精液分析參數及體外受精的臨床結果。 結果：精蟲異常病患與精蟲正常病患相比較，則出現較高之精蟲DNA碎片化發生率（18.8 % vs. 2.8 %），粒線體功能缺陷發生率（24.9 % vs. 5.7 %）與染色體倍數異常發生率（0.12 % vs. 0.06 %）。當精蟲DNA碎片化發生率超過10 % 時會影響受精率，精蟲碎片化DNA也可能影響胚胎品質，精蟲DNA碎片化發生率與精蟲速率呈負相關現象，在達到平均路徑速率（VAP）30微米/秒 與直線速率（VSL）20微米/秒 時精蟲DNA碎片化應該不會超過3 % 與正常精蟲速率相似。 結論：精蟲異常病患含有較高的DNA碎片化，粒線體功能缺陷與染色體倍數異常的機率。精蟲DNA碎片化發生率與受精率，VAP及VSL都呈現負相關，但不會影響懷孕率；可以考慮將VAP及VSL當成ICSI篩選精蟲的參考標準，至於精蟲形態則不宜做為篩選標準。總之，具有完整DNA的精蟲才能傳遞正確的遺傳訊息給下一代，而受精成功是生命誕生的第一步。品質極差的精蟲是施行ICSI的適應症，所以，遺傳諮詢及如何篩檢品質優良的精蟲就成為重要的課題。 Purpose：This study determined the incidence of sperm nuclear DNA fragmentation, mitochondrial dysfunction, and chromosomal aneuploidy. The results were correlated with the semen analysis parameters and fertilization rates. Another study evaluated sperm DNA fragmentation, in correlation with IVF/ICSI outcomes. Materials & Methods：Semen samples from 10 men showing oligoastheno- teratozoospermia（OAT）and undergoing ICSI treatment were analyzed. Another semen samples from 10men showing normozoospermia and undergoing IVF treatment were analyzed for comparison. The samples were prepared using a two-step discontinuous Percoll gradient （80%-50%）and analyzed using a Hamilton-Thorne Integrated Visual Optical System（IVOS）Sperm Analyzer. DNA fragmentation was detected with a terminal deoxynucleotidyl transferase-mediated dUTP nick end label（TUNEL）assay. Functional integrity of mitochondria was detected using an ApoalertTM Mitochondrial Membrane Sensor Kit. Chromosomal aneuploidy was assayed by fluorescence in situ hybridization. Furthermore, we collected 303 semen samples of IVF patients with or without ICSI and detected the percentage of sperm with DNA fragmentation in correlation with semen analysis parameters and IVF/ICSI outcomes. Results：A higher sperm DNA fragmentation rate（18.8 % vs. 2.8 %）, mitochondrial dysfunction rate（24.9 % vs. 5.7 %）, and chromosomal aneuploidy rate（0.12 % vs. 0.06 %）were found in the oligoasthenoterato- zoospermic patients in comparison with the normozoospermic patients. When sperm DNA fragmentation is greater than 10 %, fertilization rates are affected. Sperm DNA fragmentation may also affect embryo quality. Sperm DNA fragmentation rates were negatively correlated with velocity. To achieve a normal velocity of average path （VAP）（>30 μm/sec）and velocity of straight line（VSL）（>20 μm/sec）, sperm DNA fragmentation should not be greater than 3 %. Conclusions：The result indicates that spermatozoa from oligoathenoterato- zoospermic patients contain greater DNA fragmentation, mitochondrial dysfunction, and chromosomal aneuploidy. Sperm DNA fragmentation rates negatively correlated with fertilization rates, VAP and VSL, but did not affect pregnancy rates. Sperm DNA integrity is essential for accurate transmission of genetic material to offspring. Because extremely poor semen samples are the indication for ICSI treatment, the selection of good quality sperm for oocyte injection is very important.
