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Journal of Central South University Medical Sciences logoLink to Journal of Central South University Medical Sciences
. 2022 Jan 28;47(1):63–71. [Article in Chinese] doi: 10.11817/j.issn.1672-7347.2022.210021

精子DNA断裂指数对补救ICSI结局的影响及补救ICSI的临床价值

Influence of the sperm DNA fragmentation index on the outcome of rescue ICSI and the clinical value of rescue ICSI

CHEN Qigui 1,2, LI Dawen 1,✉, CHENG Junping 1, XUE Lintao 1, LI Jinyan 2
Editor: 郭 征
PMCID: PMC10930484  PMID: 35545364

Abstract

Objective

As a remedy for the failure of in vitro fertilization (IVF), rescue intracytoplasmic sperm injection (R-ICSI) has been widely carried out, but it has failed to significantly improve the fertilization rate and clinical pregnancy rate. Sperm DNA fragmentation index (DFI) was highly correlated with pregnancy outcome of artificial assisted reproduction. This study aims to investigate the effect of the sperm DFI on the outcome of R-ICSI and the clinical value of R-ICSI.

Methods

This retrospective analysis was conducted among 140 infertile couples receiving R-ICSI in from January 2014 to December 2019. The subjects were assigned into a total fertilization failure (TFF)+low DFI group (R-ICSI after TFF and DFI<30%) (n=63), a TFF+high DFI group (R-ICSI after TFF and DFI≥30%) (n=16), a partial fertilization failure (PFF)+low DFI group (R-ICSI after PFF and DFI<30%) (n=52), a PFF+high DFI group (R-ICSI after PFF and DFI≥30%) (n=9). All transferred embryos were come from R-ICSI. The general clinical data [infertility duration, male age, female age, basal serum level of follicle stimulating hormone (FSH), basal serum level of luteinizing hormone (LH), antral follicle count, endometrial thickness of human chorionic gonadotropin (HCG) day, and eggs] and R-ICSI cycle outcomes (fertilization rate, normal fertilization rate, cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate and live birth rate) were analyzed. In addition, the effect of R-ICSI on the fertilization outcome of conventional IVF total fertilization failure and partial fertilization failure was explored.

Results

There was no significant difference in the general clinical data and R-ICSI cycle outcome between the TFF+low DFI group and the TFF+high DFI group (all P>0.05). There was no significant difference in the general clinical data between the PFF+low DFI group and the PFF+high DFI group (all P>0.05). The fertilization rate and normal fertilization rate in the PFF+low DFI group were significantly higher than those in the PFF+high DFI group (85.40% vs 72.41%, 71.90% vs 58.62%, respectively; both P<0.05). However, there was no significant difference in cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate, and live birth rate between the 2 groups (all P>0.05). The R-ICSI cycle of TFF: A total of 79 fresh cycles, 57 fresh transplant cycles, a total of 761 unfertilized oocytes, and 584 M II oocytes were treated with R-ICSI, the fertilization rate was 83.22%, the normal fertilization rate was 75.51%, the cleavage rate was 98.15%, the good embryo rate was 40.74%, the implantation rate was 30.56%, and the clinical pregnancy rate was 43.86%; 29 live births were obtained. The R-ICSI cycle of PFF: A total of 61 fresh cycles, 31 fresh transplant cycles, a total of 721 unfertilized oocytes, and 546 M II oocytes were treated with R-ICSI; the fertilization rate was 83.33%, the normal fertilization rate was 69.78%, the cleavage rate was 97.36%, the good embryo rate was 44.39%, the implantation rate was 25.42%, and the clinical pregnancy rate was 45.16%; 12 live births were obtained.

Conclusion

In the case of partial fertilization failure of IVF, the sperm DFI affects the fertilization rate and normal fertilization rate of R-ICSI; whether it is a TFF of IVF or PFF of IVF, ICSI can be used as an effective remedy way.

Keywords: sperm DNA fragmentation index, rescue intracytoplasmic sperm injection, total fertilization failure, partial fertilization failure, outcome

辅助生殖技术(assisted reproduction technique，ART)是治疗不孕不育症的重要措施。在ART中，第一代试管婴儿技术体外受精-胚胎移植(in vitro fertilization-embryo transfer，IVF-ET)已帮助无数不孕不育症夫妇实现了生育梦想。虽然IVF技术在不断改进，但发生部分受精失败(partial fertilization failure，PFF)或完全受精失败(total fertilization failure，TFF)的风险仍然存在，其中TFF的发生率为3%~5%[1]，这会导致患者无可移植胚胎。针对此种情况，补救卵胞浆内单精子注射术(rescue intracytoplasmic sperm injection，R-ICSI)作为体外受精失败的补救方案被学者们提出，目前已在多家生殖中心实施，但也有研究[2]指出R-ICSI未能明显增加受精率、临床妊娠率，不能改善最终妊娠结局。

全世界不孕不育症夫妇数量在不断增加，其中由男性因素导致的不孕不育症约占40%，而引起男性不育的一个重要原因是精子DNA损伤[3]。目前相关研究[4-5]表明：精子DNA损伤与男性生育力下降、反复流产及人工助孕失败等高度相关，可根据精子DNA损伤情况预测妊娠结局。已有研究[6-7]指出，精子DNA断裂指数(DNA fragmentation index，DFI)与宫腔内人工授精(intrauterine insemination，IUI)、IVF和ICSI的受精率、优质胚胎率和临床妊娠率等相关。但是，DFI对R-ICSI结局影响的研究尚缺乏。因此，本研究比较不同DFI对IVF受精失败后行R-ICSI结局的影响，以探讨R-ICSI的可行性及临床价值。

1. 对象与方法

1.1. 对象

回顾性分析2014年1月至2019年12月期间在广西壮族自治区人民医院生殖医学与遗传中心IVF受精失败后行R-ICSI的140对不孕不育症夫妇。入选标准：1)女方年龄<38岁，且基础性激素水平及B超检查示卵巢功能正常；2)精子均为丈夫通过手淫获取；3)夫妇双方染色体核型正常；4)超排卵用药方案为长方案；5)新鲜移植周期的胚胎来源均为R-ICSI。排除标准：1)男方服用了影响精子质量的药物；2)女方患有子宫腺肌病、子宫内膜异位症、多囊卵巢综合征；3)夫妇双方中任意一方有生殖器发育异常、生殖系统感染等。将140对不孕不育症夫妇分为4组：1)TFF后行R-ICSI且DFI<30%为TFF+低DFI组(n=63)；2)TFF后行R-ICSI且DFI≥30%为TFF+高DFI组(n=16)；3)PFF后行R-ICSI且DFI<30%为PFF+低DFI组(n=52)；4)PFF后行R-ICSI且DFI≥30%为PFF+高DFI组(n=9)。

1.2. 药物和试剂

达菲林(注射用醋酸曲普瑞林)为益普生法国生物技术公司产品；果纳芬(注射用重组人促卵泡激素)和人绒毛膜促性腺激素(human chorionic gonadotropin，HCG)为瑞士雪兰诺公司产品；人绝经期促性腺激素(human menopausal gonadotropin，HMG)为珠海丽珠集团丽宝生物化学制药有限公司产品；瑞氏-姬姆萨染色试剂为珠海贝索生物技术有限公司产品；精子DNA完整性检测试剂盒购自西班牙Halotech公司。

1.3. 方法

1.3.1. 促排卵方案

采用长方案超促排卵，即月经周期第20~22天使用达菲林进行降调节直至HCG日止，降调节14 d后开始使用果纳芬和/或HMG促排卵，具体需视患者的激素水平变化、卵泡大小与卵泡数目等情况而定，用果纳芬和/或HMG 4 d后开始采用B超监测卵泡发育，当有2个卵泡直径大于18 mm时，于当晚21:00左右予HCG 10 000 U肌肉注射，给药36 h后于手术室行阴道超声引导下经阴道穿刺取卵。

1.3.2. 精液采集

男方禁欲2~7 d，采用手淫法取精液置于干燥消毒量杯内，标本在被转送到实验室的过程中保持20~37 ℃，置于37 ℃恒温箱内液化。

1.3.3. 精子DFI

采用精子染色质扩散试验检测精子DFI。在显微镜(×400)下计数200条以上精子，配合血细胞计数器进行人工判别，按照精子染色晕圈直径与精子核直径的比值将精子晕圈分为5个等级：大晕圈(>1)、中等晕圈(1/3~1)、小晕圈(<1/3)、无晕圈、精子头部不着色。计算每个等级的百分比，DFI为小晕圈、无晕圈、精子头部不着色3个等级的百分比之和。

1.3.4. 受精、卵裂、胚胎移植及妊娠情况

以第二极体作为早期受精的判断标准，若授精 6 h后患者所有成熟卵子中出现明确第二极体的比 例<30%，则建议行R-ICSI。对未排出第二极体的卵母细胞，在授精后6~8 h行R-ICSI治疗。R-ICSI后第1天观察卵子受精情况，以双原核(2PN)卵子作为正常受精卵。第3天按照Peter分级标准选择优质胚胎进行移植，移植胚胎数≤3个，剩余III级以上胚胎冷冻保存，并同时予黄体支持。

1.3.5. 妊娠判定及随访

移植胚胎后第14天测血β-HCG水平，若β-HCG阳性则于第28、42天用B超检查胚胎发育情况，若B超见宫腔内孕囊及心管博动则确立临床妊娠。黄体支持自妊娠8周起逐渐减量至妊娠10周停用，随访至妊娠12周转入产科门诊建档立卡及定期产检，电话随访并记录妊娠结局。

1.3.6. 结局指标

结局指标包括受精率(受精卵子数/M II期卵子数)、正常受精率(2PN卵子数/M II期卵子数)、卵裂率(卵裂数/受精卵子数)、优质胚胎率(优质胚胎数/2PN卵裂数)、种植率(孕囊数/移植胚胎数)、临床妊娠率(临床妊娠例数/移植周期例数)、活产率(活婴分娩数/移植周期例数)。

1.4. 统计学处理

采用SPSS 22.0统计软件分析数据。计量资料进行正态性检验，若符合正态分布，用均数±标准差( x¯ ±s)表示，2组比较采用t检验；若不符合正态分布，用中位数(第1百分位数，第3百分位数)[M(P 25，P 75)]表示，2组比较采用Mann-Whitney U检验。计数资料采用例(%)表示，用χ2检验。P<0.05为差异有统计学意义。

2. 结 果

2.1. TFF的R-ICSI周期不同DFI组一般情况的比较

TFF+低DFI组和TFF+高DFI组患者的不育年限、男方年龄、女方年龄、基础促卵泡素(folliclestimulating hormone，FSH)水平、基础黄体生成素(luteinizing hormone，LH)水平、窦卵泡个数、HCG日子宫内膜厚度、获卵数差异均无统计学意义(均P>0.05，表1)。

表1.

常规IVF完全受精失败R-ICSI周期不同DFI组一般情况

Table 1 General clinical data of different DFI groups in the R-ICSI cycles of total fertilization failure in conventional IVF

组别 n 不育年限 男方年龄/岁 女方年龄/岁 基础FSH/(U·L-1)
TFF+低DFI 63 3.33(1.00, 12.00) 34.00(28.00, 48.00) 32.00(25.00, 37.00) 6.82(1.56, 15.06)
TFF+高DFI 16 3.00(2.00, 12.00) 35.50(26.00, 48.00) 32.50(21.00, 37.00) 5.84(4.70, 11.33)
U 454.0 417.5 489.0 392.0
P 0.537 0.290 0.854 0.172
组别 基础LH/(U·L-1) 窦卵泡个数 HCG日子宫内膜厚度/mm 获卵个数
TFF+低DFI 3.88(1.07, 42.25) 12.00(3.00, 29.00) 10.50(7.50, 16.00) 9.00(1.00, 25.00)
TFF+高DFI 3.70(2.00, 6.67) 14.50(6.00, 20.00) 11.50(7.50, 18.00) 10.00(1.00, 26.00)
U 460.0 401.0 402.5 493.0
P 0.591 0.208 0.214 0.893
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R-ICSI：补救卵胞浆内单精子注射术；IVF：体外受精；DFI：精子DNA断裂指数；FSH：促卵泡素；LH：黄体生成素；HCG：人绒毛膜促性腺激素；TFF：完全受精失败。

2.2. DFI对常规IVF TFF的R-ICSI周期结局的影响

TFF+低DFI组和TFF+高DFI组患者行R-ICSI时受精率、正常受精率、卵裂率、优质胚胎率、种植率、临床妊娠率及活产率差异均无统计学意义(均P>0.05，表2)。

表2.

常规IVF完全受精失败的R-ICSI周期不同DFI组的结局

Table 2 Outcome of different DFI groups in the R-ICSI cycles of total fertilization failure in conventional IVF

组别 n 受精率/% 正常受精率/% 卵裂率/% 优质胚胎率/% 种植率/% 临床妊娠率/% 活产率/%
TFF+低DFI 63 82.64 75.16 98.40 40.30 32.58 46.81 42.55
TFF+高DFI 16 85.27 76.74 97.27 42.27 21.05 30.00 20.00
χ2 0.499 0.136 0.139 0.121 0.981 0.387 0.946
P 0.480 0.713 0.710 0.728 0.322 0.534 0.331
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R-ICSI：补救卵胞浆内单精子注射术；IVF：体外受精；DFI：精子DNA断裂指数；TFF：完全受精失败。

2.3. 常规IVF PFF的R-ICSI周期不同DFI组一般 情况的比较

PFF+低DFI组和PFF+高DFI组患者的不育年限、男方年龄、女方年龄、基础FSH、基础LH、窦卵泡个数、HCG日子宫内膜厚度、获卵数差异均无统计学意义(均P>0.05，表3)。

表3.

常规IVF部分受精失败R-ICSI周期不同DFI组一般情况

Table 3 General clinical data of different DFI groups in the R-ICSI cycles of partial fertilization failure in conventional IVF

组别 n 不育年限 男方年龄/岁 女方年龄/岁 基础FSH/(U·L-1)
PFF+低DFI 52 3.00(0.17, 11.58) 34.92±5.11 32.00(23.00, 37.00) 6.35±1.46
PFF+高DFI 9 4.25(1.00, 8.00) 32.44±5.77 29.00(23.00, 37.00) 6.88±1.70
t/U 199.0 1.319 162.5 -0.990
P 0.471 0.192 0.144 0.326
组别 基础LH/(U·L-1) 窦卵泡个数 HCG日子宫内膜厚度/mm 获卵个数
PFF+低DFI 4.17(1.65, 10.14) 15.87±6.75 11.26±2.46 13.00(6.00, 27.00)
PFF+高DFI 4.89(2.07, 6.98) 15.56±6.11 11.41±2.54 14.00(9.00, 30.00)
t/U 220.0 0.129 -0.165 192.0
P 0.776 0.898 0.869 0.392
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R-ICSI：补救卵胞浆内单精子注射术；IVF：体外受精；DFI：精子DNA断裂指数；FSH：促卵泡素；LH：黄体生成素；HCG：人绒毛膜促性腺激素；PFF：部分受精失败。

2.4. DFI对常规IVF PFF的R-ICSI周期结局的影响

PFF+低DFI组和PFF+高DFI组患者受精率、正常受精率差异均有统计学意义(均P<0.05)；2组患者卵裂率、优质胚胎率、种植率、临床妊娠率及活产率差异均无统计学意义(均P>0.05，表4)。

表4.

常规IVF部分受精失败R-ICSI周期不同DFI组的结局

Table 4 Outcome of different DFI groups in the R-ICSI cycles of partial fertilization failure in conventional IVF

组别 n 受精率/% 正常受精率/% 卵裂率/% 优质胚胎率/% 种植率/% 临床妊娠率/% 活产率/%
PFF+低DFI 52 85.40 71.90 97.70 45.23 27.27 48.28 37.93
PFF+高DFI 9 72.41 58.62 95.24 38.78 0 0 0
χ2 8.885 6.112 0.504 0.719
P 0.003 0.013 0.478 0.397 0.564 0.488 0.527
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R-ICSI：补救卵胞浆内单精子注射术；IVF：体外受精；DFI：精子DNA断裂指数；PFF：部分受精失败。

2.5. 常规IVF R-ICSI周期胚胎发育及妊娠结局

常规IVF TFF的R-ICSI周期(新鲜移植周期的胚胎来源均为R-ICSI)：共79个新鲜周期，其中57个新鲜移植周期，22个取消周期，72.15%(57/79)的周期获得移植；共761个未受精卵，对584个M II期卵子行R-ICSI，卵子再利用率为76.74%(584/761)。受精卵子数为486个，2PN受精卵子数为441个，卵裂数为477个，2PN卵裂数为432个，优质胚胎数为176个。受精率为83.22%(486/584)，正常受精率为75.51%(441/584)，卵裂率为98.15%(477/486)，优质胚胎率为40.74%(176/432)。总共移植108枚胚胎，孕囊数33个，种植率为30.56%(33/108)。临床妊娠25例，临床妊娠率为43.86%(25/57)。共22例患者分娩，29个活产儿(单胎15个，双胎7个)，3例流产。

常规IVF PFF的R-ICSI周期(新鲜移植周期的胚胎来源均为R-ICSI)：共61个新鲜周期，其中31个新鲜移植周期，30个取消周期，50.82%(31/61)的周期获得移植；共721个未受精卵，对546个M II期卵子行R-ICSI，卵子再利用率为75.73%(546/721)。受精卵子数为455个，2PN受精卵子数为381个，卵裂数为443个，2PN卵裂数为374个，优质胚胎数为166个。受精率为83.33%(455/546)，正常受精率为69.78%(381/546)，卵裂率为97.36%(443/455)，优质胚胎率为44.39%(166/374)。总共移植59枚胚胎，孕囊数15个，种植率为25.42%(15/59)。临床妊娠14例，临床妊娠率为45.16%(14/31)。共11例患者分娩，12个活产儿(单胎10个，双胎1个)，1例流产，2例异位妊娠。

3. 讨 论

R-ICSI为常规IVF周期失败的不孕不育症夫妇提供了再次受孕的可能，也为生殖医师提供了除取消当前治疗周期方法之外的另一种方案。R-ICSI可以绕过机械屏障帮助精子穿透卵子，但不能规避受精过程中遇到的所有问题。对于精子与卵子透明带结合失败所导致的受精失败，R-ICSI是可行的；但是，如果是由于随后的卵母细胞激活和原核形成过程中出现障碍导致受精失败，R-ICSI失败的可能性则会增加。研究[8]指出：当IVF受精率低于25%时，R-ICSI是一种有效的补救方案；而当IVF受精率>25%时，则不建议实施R-ICSI。因为相对于取消当前周期，实施R-ICSI手术具有更大的风险和较低的回报。目前应用较为广泛的有早期R-ICSI和晚期R-ICSI两种方案。有学者[9]对这两种方案进行了比较，认为晚期R-ICSI在改善胚胎发育及临床妊娠结局方面不如早期R-ICSI，在常规IVF受精失败或受精率低下的情况下应首选早期R-ICSI。

众所周知，高质量胚胎的产生需要高质量的配子，精子与卵母细胞相互作用障碍是导致ART失败的主要因素。男方精子DNA异常会对受精、卵裂、受精卵着床、胚胎植入等结局产生不利影响[10]。在精子生成的过程中，男性的患病情况和自身状态、阴部温度、吸烟、酗酒、年龄及外界环境等因素均可损伤精子DNA[11]。但是，目前没有足够的证据支持BMI与精子DNA断裂率存在关联[12]。精子DNA损伤机制包括精子发生异常、氧化应激损伤、精子凋亡异常、染色质异常等。染色质组装异常可造成精子DNA损伤，是影响男性生殖能力及受精的主要原因[13]。学者们已对精子DFI与IVF/ICSI结局的关系进行了大量的研究。蔡文伟等[14]研究指出，精子DNA断裂率与IVF受精率呈显著负相关。汪李虎等[15]探讨了精子DNA损伤与IVF临床结局的关系，发现药物治疗可提高精子DNA的完整性，从而改善优质胚胎率、种植率及妊娠率。精子DNA损伤会增加ICSI周期流产风险，降低胚胎质量，导致着床反复失败，精子DFI可作为ART结局的预测因素[16]。在ART中，随着精子DFI的增加，活产率显著下降，两者呈负相关[17]。精子DFI的增加会影响胚胎正常发育、受精卵着床，甚至影响ICSI妊娠结局。但是也有学者认为精子DFI与IVF/ICSI结局无相关性。Green等[18]研究指出，DFI>15%组与DFI≤15%组的受精率、囊胚形成率、临床妊娠率和流产率差异无统计学意义，提出DFI不能预测受精率、胚胎质量和临床妊娠率。Sun等[19]研究精子DFI对IVF/ICSI周期的影响，将其分为DFI≤30%和DFI>30%组，发现精子DFI对IVF/ICSI周期的受精率、优质胚胎率、临床妊娠率无影响。存在差别的原因可能是：1)不同研究纳入标准与排除标准不一；2)实验室方法不同；3)精子DFI检测所用方法不同等。肖鑫等[20]指出：当精子密度<11.1×106/mL，正常形态率<1.4%，DFI>5.9%，顶体完整 率<77.3%，顶体反应率<15.7%时，IVF-ET并非治疗不孕不育症的第一选择。

国内外对男性精子DFI对R-ICSI结局影响的研究尚少，且纳入及排除标准不尽相同，因此，在本研究中，我们首先将患者分为TFF后行R-ICSI和PFF后行R-ICSI，再分别比较DFI<30%及DFI≥30%患者的一般情况和R-ICSI结局，填补了以往研究的不足。结果发现：在TFF后行R-ICSI的患者中，DFI<30%组及DFI≥30%组的一般情况、胚胎发育情况及临床结局差异均无统计学意义。这与何泳志等[21]的研究结论一致，即精子DFI与R-ICSI周期的受精率、卵裂率、优质胚胎率不相关。另外，本研究还增加了正常受精率、种植率、临床妊娠率及活产率这些结局指标，结果发现精子DFI对这些指标也无明显影响。分析原因可能为：受精是精子与卵子识别、结合、融合和激活的过程，涉及一系列蛋白质反应和蛋白质的相互作用，精子和卵子的异常及二者之间相互作用蛋白质的缺失或功能异常均可导致受精失败[22]。Saleh等[23]研究报道精子PLCζ蛋白质缺失是导致受精失败和钙离子震荡频率和幅度变化的重要原因。发生TFF可能与精子、卵子自身功能缺陷有关，如精卵膜识别与融合蛋白质缺失、精卵融合缺陷、卵子钙离子震荡不足及精子诱导钙离子信号低下等因素就有可能使精子入卵后减数分裂、精子核去致密失败，从而导致受精失败，而与精子DNA损伤无显著关系[24]。从基因发生机制角度分析，在TFF中，精卵结合异常可能与基因变异导致的功能蛋白质表达异常有关[25]。另外，也可能与样本量不足或受精子形态、顶体、卵子质量、活性氧水平及不良生活习惯等其他因素影响有关[26]。

在R-ICSI(PFF)中，DFI<30%的受精率、正常受精率大于DFI≥30%，两者比较差异有统计学意义。这与既往研究[27]认为低DFI组比高DFI组通常有更好的胚胎发育结局一致。精子DNA是重要的遗传物质，其染色质结构的正常与否对精子的授精能力、正常胚胎发育起重要作用。精子DNA损伤愈少，授精能力一般越好。妊娠前应行精子DNA检测，为避免IVF受精率欠佳的情况，建议当DFI>15.5%时行R-ICSI辅助生殖[28]。但是，在本研究中，PFF+低DFI组和PFF+高DFI组的种植率、临床妊娠率及活产率差异无统计学意义。这提示精子DFI对早期胚胎发育有影响，而对后续的临床妊娠结局无显著影响，与Simon等[29]的研究结论一致。黄文思等[30]指出精子DFI影响受精率、卵裂率等早期胚胎发育指标，但对临床妊娠结局无影响。精子DNA损伤对体外受精早期胚胎发育产生负面影响[31]。在最初与卵母细胞结合时，早期胚胎发育由男性精子因素主导，但随着进程深入，由于卵母细胞存在主动修复机制[32]，对精子DNA进行修复，最终胚胎发育仍可正常，因而出现了不同损伤程度精子DNA的临床妊娠结局差异无统计学意义。

常规IVF受精失败后行R-ICSI方案可避免周期取消，已被广泛应用于生殖学领域。本研究探讨了R-ICSI的临床价值，并对IVF TFF或PFF进行细化分类，发现在IVF TFF和PFF中，R-ICSI均可获得不错的妊娠结局，分别获得29个和12个活产儿。这与国内外的研究结论一致，欧建平等[33]、Huang等[34]的研究表明：常规IVF TFF后行R-ICSI可降低周期取消率，取得较好的受精率、卵裂率及妊娠率。曾勇等[35]也对IVF TFF或PFF进行了分类分析，结果表明R-ICSI是一种不错的IVF受精失败的补救方法，但与之相比，本研究的数据更充足，R-ICSI相关结局指标更多、更全面，并进一步证明了无论是对TFF还是PFF，均可推荐应用R-ICSI。R-ICSI不仅可避免周期取消，给患者再次受孕的希望，而且其胚胎发育及临床结局可为下一周期的辅助生殖方案提供建设性的参考指标。综上，笔者认为精子DFI在男性生育能力的评估中非常有价值，可在一定程度上影响早期胚胎发育，同时R-ICSI可作为IVF受精失败后的推荐补救方案。

基金资助

国家自然科学基金(81360107)；广西壮族自治区自然科学基金(2019GXNSFAA185056)；广西壮族自治区卫生健康委员会科研项目(Z20170376)；南宁市青秀区重点研发计划(2020030)。

This work was supported by the National Natural Science Foundation (81360107), the Guangxi Natural Science Foundation (2019GXNSFAA185056), the Scientific Research Project of Guangxi Health Commission (Z20170376), and the Key Research and Development Program of Nanning Qingxiu District (2020030), China.

利益冲突声明

作者声称无任何利益冲突。

作者贡献

陈其桂 实验设计与实施研究，数据分析，撰写论文。李大文，成俊萍 实验设计，论文审阅及指导，经费支持。薛林涛 实验设计与指导，经费支持。李金燕 数据采集和分析。

原文网址

http://xbyxb.csu.edu.cn/xbwk/fileup/PDF/20220163.pdf

参考文献

  • 1. Yu M, Zhao H, Chen T, et al. Mutational analysis of IZUMO1R in women with fertilization failure and polyspermy after in vitro fertilization[J]. J Assist Reprod Genet, 2018, 35(3): 539-544. 10.1007/s10815-017-1101-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Beck-Fruchter R, Lavee M, Weiss A, et al. Rescue intracytoplasmic sperm injection: a systematic review[J]. Fertil Steril, 2014, 101(3): 690-698. 10.1016/j.fertnstert.2013.12.004. [DOI] [PubMed] [Google Scholar]
  • 3. Hanson BM, Kaser DJ, Franasiak JM. Male infertility and the future of in vitro fertilization[J]. Urol Clin North Am, 2020, 47(2): 257-270. 10.1016/j.ucl.2019.12.012. [DOI] [PubMed] [Google Scholar]
  • 4. Sakkas D, Manicardi GC, Bizzaro D. Sperm nuclear DNA damage in the human[J]. Adv Exp Med Biol, 2003, 518: 73-84. 10.1007/978-1-4419-9190-4_7. [DOI] [PubMed] [Google Scholar]
  • 5. Shafik A, Shafik AA, Shafik I, et al. Sperm DNA fragmentation[J]. Arch Androl, 2006, 52(3): 197-208. 10.1080/01485010500503561. [DOI] [PubMed] [Google Scholar]
  • 6. Siddhartha N, Reddy NS, Pandurangi M, et al. The effect of sperm DNA fragmentation index on the outcome of intrauterine insemination and intracytoplasmic sperm injection[J]. J Hum Reprod Sci, 2019, 12(3): 189-198. 10.4103/jhrs.JHRS_22_19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Oleszczuk K, Giwercman A, Bungum M. Sperm chromatin structure assay in prediction of in vitro fertilization outcome[J]. Andrology, 2016, 4(2): 290-296. 10.1111/andr.12153. [DOI] [PubMed] [Google Scholar]
  • 8. Cao S, Wu X, Zhao C, et al. Determining the need for rescue intracytoplasmic sperm injection in partial fertilisation failure during a conventional IVF cycle[J]. Andrologia, 2016, 48(10): 1138-1144. 10.1111/and.12551. [DOI] [PubMed] [Google Scholar]
  • 9. 孙凯, 王建业, 唐宁. 早期或晚期R-ICSI对IVF-ET早期胚胎发育和临床妊娠结局的影响[J]. 中国优生与遗传杂志, 2017, 25(9): 103-106. 10.13404/j.cnki.cjbhh.2017.09.041. [DOI] [Google Scholar]; SUN Kai, WANG Jianye, TANG Ning. Effect of early or late re-ICSI on early embryo development and clinical pregnancy results in IVF-ET[J]. Chinese Journal of Birth Health & Heredity, 2017, 25(9): 103-106. 10.13404/j.cnki.cjbhh.2017.09.041. [DOI] [Google Scholar]
  • 10. Shamsi MB, Kumar R, Dada R. Evaluation of nuclear DNA damage in human spermatozoa in men opting for assisted reproduction[J]. Indian J Med Res, 2008, 127(2): 115-123. https://pubmed.ncbi.nlm.nih.gov/18403788/. [PubMed] [Google Scholar]
  • 11. Rubes J, Selevan SG, Evenson DP, et al. Episodic air pollution is associated with increased DNA fragmentation in human sperm without other changes in semen quality[J]. Hum Reprod, 2005, 20(10): 2776-2783. 10.1093/humrep/dei122. [DOI] [PubMed] [Google Scholar]
  • 12. Sepidarkish M, Maleki-Hajiagha A, Maroufizadeh S, et al. The effect of body mass index on sperm DNA fragmentation: a systematic review and meta-analysis[J]. Int J Obes (Lond), 2020, 44(3): 549-558. 10.1038/s41366-020-0524-8. [DOI] [PubMed] [Google Scholar]
  • 13. O'Hagan HM. Chromatin modifications during repair of environmental exposure-induced DNA damage: a potential mechanism for stable epigenetic alterations[J]. Environ Mol Mutagen, 2014, 55(3): 278-291. 10.1002/em.21830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. 蔡文伟, 莫敦胜, 江楠, 等. 男性不育患者精子DNA完整性与精浆氧化应激水平的关系及其对体外受精的影响[J]. 中华男科学杂志, 2016, 22(10): 892-896. 10.13263/j.cnki.nja.2016.10.006. [DOI] [Google Scholar]; CAI Wenwei, MO Dunsheng, JIANG Nan, et al. Association of sperm DNA integrity with seminal plasma oxidative stress and its influence on in vitro fertilization in infertile males[J]. National Journal of Andrology, 2016, 22(10): 892-896. 10.13263/j.cnki.nja.2016.10.006. [DOI] [PubMed] [Google Scholar]
  • 15. 汪李虎, 汪艳, 梁嘉颖, 等. 精子DNA完整性对体外授精-胚胎移植妊娠结局的影响[J]. 中国男科学杂志, 2016, 30(10): 38-41. 10.3969/j.issn.1008-0848.2016.10.008. [DOI] [Google Scholar]; WANG Lihu, WANG Yan, LIANG Jiaying, et al. Influence of the sperm DNA integrity on the clinical pregnancy outcomes of in vitro fertilization and embryo transfer[J]. Chinese Journal of Andrology, 2016, 30(10): 38-41. 10.3969/j.issn.1008-0848.2016.10.008. [DOI] [Google Scholar]
  • 16. Garolla A, Cosci I, Bertoldo A, et al. DNA double strand breaks in human spermatozoa can be predictive for assisted reproductive outcome[J]. Reprod Biomed Online, 2015, 31(1): 100-107. 10.1016/j.rbmo.2015.03.009. [DOI] [PubMed] [Google Scholar]
  • 17. Osman A, Alsomait H, Seshadri S, et al. The effect of sperm DNA fragmentation on live birth rate after IVF or ICSI: a systematic review and meta-analysis[J]. Reprod Biomed Online, 2015, 30(2): 120-127. 10.1016/j.rbmo.2014.10.018. [DOI] [PubMed] [Google Scholar]
  • 18. Green KA, Patounakis G, Dougherty MP, et al. Sperm DNA fragmentation on the day of fertilization is not associated with embryologic or clinical outcomes after IVF/ICSI[J]. J Assist Reprod Genet, 2020, 37(1): 71-76. 10.1007/s10815-019-01632-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Sun TC, Zhang Y, Li HT, et al. Sperm DNA fragmentation index, as measured by sperm chromatin dispersion, might not predict assisted reproductive outcome[J]. Taiwan J Obstet Gynecol, 2018, 57(4): 493-498. 10.1016/j.tjog.2018.06.003. [DOI] [PubMed] [Google Scholar]
  • 20. 肖鑫, 李大文, 冯刚, 等. 精子质量对不孕不育患者辅助生殖技术选择的指导作用[J]. 山东医药, 2016, 56(4): 19-21. 10.3969/j.issn.1002-266X.2016.04.006. [DOI] [Google Scholar]; XIAO Xin, LI Dawen, FENG Gang, et al. Guidance of sperm quality in choosing assisted reproductive technology for infertility patients[J]. Shandong Medical Journal, 2016, 56(4): 19-21. 10.3969/j.issn.1002-266X.2016.04.006. [DOI] [Google Scholar]
  • 21. 何泳志, 李大文, 成俊萍, 等. 精子DNA完整率、精子顶体完整率及反应率对补救卵泡浆内单精子注射术的影响[J]. 南方医科大学学报, 2016, 36(1): 140-144. 10.3969/j.issn.1673-4254.2016.01.26. [DOI] [Google Scholar]; HE Yongzhi, LI Dawen, CHENG Junping, et al. Impact of sperm DNA and acrosome integrity and acrosome reaction rate on outcomes of rescue intracytoplasmic sperm injection[J]. Journal of Southern Medical University, 2016, 36(1): 140-144. 10.3969/j.issn.1673-4254.2016.01.26. [DOI] [PubMed] [Google Scholar]
  • 22. 杨沫, 杨兴雯, 李默. 精卵识别分子机制研究进展[J]. 中华生殖与避孕杂志, 2019, 39(2): 161-164. 10.3760/cma.j.issn.2096-2916.2019.02.016. [DOI] [Google Scholar]; YANG Mo, YANG Xingwen, LI Mo. Molecular mechanisms of sperm-egg recognition[J]. Chinese Journal of Reproduction and Contraception, 2019, 39(2): 161-164. 10.3760/cma.j.issn.2096-2916.2019.02.016. [DOI] [Google Scholar]
  • 23. Saleh A, Kashir J, Thanassoulas A, et al. Essential role of sperm-specific PLC-Zeta in egg activation and male factor infertility: An update[J]. Front Cell Dev Biol, 2020, 8: 28. 10.3389/fcell.2020.00028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. Sun B, Yeh J. Calcium oscillatory patterns and oocyte activation during fertilization: a possible mechanism for total fertilization failure (TFF) in human in vitro fertilization?[J]. Reprod Sci, 2021, 28(3): 639-648. 10.1007/s43032-020-00293-5. [DOI] [PubMed] [Google Scholar]
  • 25. Li M, Jia M, Zhao X, et al. A new NLRP5 mutation causes female infertility and total fertilization failure[J]. Gynecol Endocrinol, 2021, 37(3): 283-284. 10.1080/09513590.2020.1832069. [DOI] [PubMed] [Google Scholar]
  • 26. 戴菁, 黄增辉, 张硕屏, 等. 体外受精-胚胎移植中完全受精失败的原因分析[J]. 现代生物医学进展, 2017, 17(27): 5389-5391+5395. 10.13241/j.cnki.pmb.2017.27.048. 28806088 [DOI] [Google Scholar]; DAI Jing, HUANG Zenghui, ZHANG Shuoping, et al. Analysis of the causes of total fertilization failure in vitro fertilization-embryo transfer[J]. Progress in Modern Biomedicine, 2017, 17(27): 5389-5391+5395. 10.13241/j.cnki.pmb.2017.27.048. [DOI] [Google Scholar]
  • 27. Borges E, Zanetti BF, Setti AS, et al. Sperm DNA fragmentation is correlated with poor embryo development, lower implantation rate, and higher miscarriage rate in reproductive cycles of non-male factor infertility[J]. Fertil Steril, 2019, 112(3): 483-490. 10.1016/j.fertnstert.2019.04.029. [DOI] [PubMed] [Google Scholar]
  • 28. 雷承泳, 罗朝霞, 赖锦锋. 精子质量对不孕不育患者辅助生殖技术选择的指导作用探究[J]. 当代医学, 2018, 24(10): 146-147. 10.3969/j.issn.1009-4393.2018.10.071. [DOI] [Google Scholar]; LEI Chengyong, LUO Zhaoxia, LAI Jinfeng. Sperm quality in patients with infertility assisted reproductive technology selection guidance to explore[J]. Contemporary Medicine, 2018, 24(10): 146-147. 10.3969/j.issn.1009-4393.2018.10.071. [DOI] [Google Scholar]
  • 29. Simon L, Murphy K, Shamsi MB, et al. Paternal influence of sperm DNA integrity on early embryonic development[J]. Hum Reprod, 2014, 29(11): 2402-2412. 10.1093/humrep/deu228 [DOI] [PubMed] [Google Scholar]
  • 30. 黄文思, 叶丽君, 宋明哲, 等. 精子DNA碎片指数与IVF/ICSI结局的相关性[J]. 生殖医学杂志, 2019, 28(10): 1195-1201. 10.3969/j.issn.1004-3845.2019.10.018. [DOI] [Google Scholar]; HUANG Wensi, YE Lijun, SONG Mingzhe, et al. Association between sperm DNA fragmentation index and IVF/ICSI outcome[J]. Journal of Reproductive Medicine, 2019, 28(10): 1195-1201. 10.3969/j.issn.1004-3845.2019.10.018. [DOI] [Google Scholar]
  • 31. Zheng WW, Song G, Wang QL, et al. Sperm DNA damage has a negative effect on early embryonic development following in vitro fertilization[J]. Asian J Androl, 2018, 20(1): 75-79. 10.4103/aja.aja_19_17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. Bazrgar M, Gourabi H, Yazdi PE, et al. DNA repair signalling pathway genes are overexpressed in poor-quality pre-implantation human embryos with complex aneuploidy[J]. Eur J Obstet Gynecol Reprod Biol, 2014, 175: 152-156. 10.1016/j.ejogrb.2014.01.010. [DOI] [PubMed] [Google Scholar]
  • 33. 欧建平, 赵伟娥, 魏思达, 等. 常规体外受精失败后行补救性卵胞质内单精子注射的临床总结[J]. 中国优生与遗传杂志, 2013, 21(10): 102-103. 10.13404/j.cnki.cjbhh.2013.10.047. [DOI] [Google Scholar]; Jianping OU, ZHAO Weie, WEI Sida, et al. Analysis of the clinical outcomes of rescue ICSI after total fertilization failure in conventional IVF[J]. Chinese Journal of Birth Health & Heredity, 2013, 21(10): 102-103. 10.13404/j.cnki.cjbhh.2013.10.047. [DOI] [Google Scholar]
  • 34. Huang B, Qian K, Li Z, et al. Neonatal outcomes after early rescue intracytoplasmic sperm injection: an analysis of a 5-year period[J]. Fertil Steril, 2015, 103(6): 1432-7.e1. 10.1016/j.fertnstert.2015.02.026. [DOI] [PubMed] [Google Scholar]
  • 35. 曾勇, 胡晓东, 宋成, 等. 体外受精失败后补行卵胞浆内单精子注射的临床价值探讨[J]. 生殖与避孕, 2006(1): 32-34+39. 10.3969/j.issn.0253-357X.2006.01.006. [DOI] [Google Scholar]; ZENG Yong, HU Xiaodong, SONG Cheng, et al. Application value of rescue ICSI of unfertilized oocytes after IVF[J]. Reproduction & Contraception, 2006(1): 32-34+39. 10.3969/j.issn.0253-357X.2006.01.006. [DOI] [Google Scholar]

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