长期过量饮酒对精子质量及生精细胞凋亡的影响

The long-term excessive alcohol consumption on sperm quality and the impact of germ cell apoptosis

目的探讨长期高浓度过量饮酒者精液中一氧化氮（NO）含量其精子质量和生精细胞凋亡与不育的关系。方法116例过量饮酒男性患者精液分为4组，30例不饮酒男性精液为对照，采用硝酸还原酶法特异地将NO代谢产物硝酸盐（NO3^-）测还原成亚硝酸盐（NO2^-）总量代表NO水平。用脱氧核苷酸末端转移酶（TdT）介导的缺口末端标记（TUNEL）法和双目光学显微镜，分别检测和观察生精细胞的凋亡率及形态结构。用SQA—V全自动精子质量分析仪，测定精子质量。结果未饮酒生育组精液中NO的含量为（54．81±11．45）μmol／L，生精细胞的凋亡率（4．52±I．23）％，精子质量活率（80．24±0．17）％、活力a＋b（78．32±0．12）％、畸形率（5．304±0．13）％，与长期高浓度过量饮酒C组[（128．83±22．73）μmol／L，（17．34±2．53）％，（51．18±0．58）％，（21．45±0．26）％，（21．12±3．24）％]相比，差异均有统计学意义（t：10．04，17．38，6．69，15．59，17．02，P〈0．01）。长期高浓度过量饮酒各组NO的含量和生精细胞的凋亡率呈正相关（r=0．93，P〈0．01）。凋亡的生精细胞核染色质浓缩于核周形成新月形，核裂解形成凋亡小体。结论长期高浓度过量饮酒者精液中NO的含量和生精细胞的凋亡率均升高，精子质量低下。提示长期高浓度过量饮酒可致机体NO过度产生而促使生精细胞凋亡致男性不育的因素之一。

Objective To investigate the long-term excessive drinking semen nitric oxide （NO） content in their sperm quality and spermatogenic cell apoptosis and infertility. Methods Nitrate reductase method was used to specific reduce the NO metabolites nitrate （ NO3 ） to nitrite （ NO2^ - ） , which was used on behalf of the total NO level. Terminal deoxynucleotidyl transferase （TdT） and mediated nick end labeling （TUNEL） method and binocular optical microscope were used to detect and observe the rate of apoptosis of spermatogenic cells and the morphological structure. The SQA-V sperm quality automatic analyzer was used to measure sperm quality. Results In not drinking semen fertility group, NO content was （54. 81 ± 11.45） μmol / L, the rate of spermatogenic cell apoptosis was （4. 52 ± 1.23 ） %, sperm motility was （ 80. 24 ± 0. 17 ） %, energy a ＋ b （78.32 ± 0. 12 ） %, deformity rate （5.30 ± 0. 13 ） %, and long-term excessive alcohol infertility C group was [（ 128.83 ± 22.73） μmol/L, （ 17.34 ±2.53）%, （51.18 ± 0. 58）%, （21.45 ±0. 26）%, （21.12 ±3.24）% ] respectively. Compared to a very significant differerlce （ t = 10.04, 17.38, 6. 69, 15.59, 17.02, P 〈0. 01 ） . In long-term excessive drinking group, the levels of NO and spermatogenesis cell apoptosis rate was significantly positive eorrelated （ r = 0. 93, P 〈 0.01 ）.Apoptosis of spermatogenic cell nucleus chromatin was condensed in the formation of crescent-shaped peri- nuclear, nuclear was cleavaged to form apoptotic bodies. Conclusions In the long-term excessive drinking semen, NO content and spermatogenic cell apoptosis rate was increased, the sperm had poor quality. The results showed that the long-term excessive drinking can cause germ cell apoptosis in male infertility and promote the body to overproduce NO, whichmay be one of the reasons for male infertility.