不同年龄对精子凋亡率及DNA完整性影响的研究

Effects of males' age on sperm apoptosis and DNA integrity

目的:探讨不同年龄阶段精子凋亡率、精子DNA完整性及精液基本参数与年龄是否存在相关性。方法:随机选取在我科因女方因素行体外受精-胚胎移植男性精液标本104例,按男方年龄分为3组,年龄<35岁组43例、35～39岁组31例、≥40岁组30例,按WHO手册第4版手工分析精液常规参数,流式细胞术(FCM)结合FITC-Annexin V/PI荧光染色检测精子凋亡率,吖啶橙荧光染色检测精子DNA完整性。结果:①3组精液量[(2.87±0.89)ml vs(2.98±1.09)ml vs(2.65±0.95)ml]、精子浓度[(60.40±25.43)×106/ml vs(69.74±28.33)×106/ml vs(55.97±27.22)×106/ml]无显著性差异(P>0.05),≥40岁组前向运动精子百分率[(39.00±8.35)%]减少,与<35岁组[(48.72±9.89)%]和35～39岁组[(45.65±10.55)%]相比,差异有显著性(P<0.01);≥40岁组正常形态精子百分率[(11.11±8.26)%]与<35岁组[(16.43±8.75)%]相比,差异有显著性(P<0.01)。②精子凋亡在不同年龄组人群均有一定发生率,≥40岁组精子凋亡率为[(11.82±5.77)%],高于<35岁组[(7.04±3.50)%]和35～39岁组[(9.42±4.73)%],与<35岁组相比结果有统计学意义(P<0.01);≥40岁组DNA完整精子百分率降低[(75.52±10.60)%],与<35岁组[(86.55±5.60)%]和35～39岁组[(81.39±8.94)%]相比,差异有显著性(P<0.01);③男性年龄与精子凋亡百分率呈正相关(P<0.01),与前向运动精子、DNA完整精子百分率呈负相关(P<0.01)。结论:男性年龄增加可能导致前向运动精子百分率减少,正常形态精子百分率降低,精子凋亡百分率增加,DNA完整精子百分率性降低,在生育过程中,男性年龄应引起关注。

Objective： To explore the correlation of males＇age with sperm apoptosis, sperm DNA integrity and other seminal pa- rameters. Methods： We collected 104 semen samples and divided them into three groups according to the males＇age： 〈 35 yr （n = 43）, 35 -39 yr （n = 31）, and ≥40 yr （n = 30）. Based on the WHO Laboratory Manual （4th ed）, we detected the seminal pa- rameters, calculated the percentage of apoptotic sperm by flow cytometry （FCM）, determined sperm DNA integrity by Acridine orange staining, and compared the results among the three groups. Results： There were no statistically significant differences among the 〈 35 yr, 35 - 39 yr and ≥40 yr groups in semen volume （ [ 2.87 ± 0.89 ] ml vs [ 2.98 ± 1.09 ] ml vs [ 2.65 ± 0.95 ] ml）, sperm con- centration （ [ 60.40 ± 25.43 ] ×106/ml vs [ 69.74 ± 28.33 ] × 106/ml vs [ 55.97 ± 27.22 ]×106/ml） （P 〉 0.05）. The percentage of progressively motile sperm was significantly lower in the 〉 40 yr （ [ 39.00 ± 8.35 ] % ） than in the 〈 35 and 35 - 39 yr groups （ [48.73 ±9.89] % and [45.65 ±10.55] % ） （P 〈0. O1 ）, and so was the percentage of morphologically normal sperm in the I〉40yr than in the 〈 35 yr group （ [ 11.11 ± 8.26 ] % vs [ 16.43 ±8.75 ] %, P 〈 0.01 ）. The percentage of apoptotic sperm was markedly higher in the 〉≥40 yr than in the 〈 35 yr group （ [ 11.82 ± 5.77 ] % vs [ 7.04 ± 3.50 ] %, P 〈 0.01 ）, while the sperm DNA integrity significantly reduced in the ≥40 yr group （ [75.52 ± 10.601% ） as compared with the 〈35 yr （ [86.55 ±5.60] ）% and 35 -39 yr group （ [ 81.39 ± 8.94 ] % ） （P 〈 O. 01 ）. The males＇age was correlated positively with the rate of sperm apoptosis （P 〈 0.01 ）, and negatively with sperm DNA integrity and the percentage of progressively motile sperm （ P 〈 0.01 ）. Conclusion ： The advance in males＇age increases sperm apoptosis and reduces sperm progressive motility, normal morphology and DNA integrity.