Zhou Wan-Hao, Ma Xu, Jiang Hui, Yuan Ren-Pei, Chen Qian, Sui Yu-Jie, Jia Meng-Chun
Postgraduate Institute of Peking Union Medical College, Beijing 100730, China.
Zhonghua Nan Ke Xue. 2008 May;14(5):391-5.
To determine the sperm mtDNA content, mtDNA4977bp deletion and ROS in the seminal plasma of normal and leukocytospermia men, and to investigate the correlation of the changes of sperm mtDNA with the increase of leukocytes and reactive oxgygen species (ROS) in the seminal plasma.
Seventy-eight semen samples from leukocytospermia patients and 31 from healthy donors were divided into 3 layers, supernatant fluid, 30% sperm and 80% sperm, by Percoll gradient centrifugation, their sperm mtDNA content and mtDNA4977bp deletion quantitatively analyzed by real-time PCR, and the level of ROS determined by flow cytometry.
The ROS in the seminal plasma and the sperm mtDNA contents of the three layers were all significantly higher in the leukocytospermia group than in the healthy control (P < 0.01). In the supernatant fluid and 80% layers, mtDNA4977bp deletion showed no obvious difference between the control and the leukocytospermia group, but was significantly higher in the 30% layer of the latter (P < 0.01). The ROS level was found positively correlated with the mtDNA content in the 30% (r = 0.347, P < 0.01) and the 80% layer (r = 0.456, P < 0.01), but not in the supernatant layer.
The increase of leukocytes and ROS may be one of the causes of the enhanced sperm mtDNA content, but has no significant impact on the mtDNA4977bp deletion.
测定正常男性和白细胞精子症男性精液中精子线粒体DNA含量、线粒体DNA 4977bp缺失及活性氧(ROS)水平,探讨精子线粒体DNA变化与精液中白细胞增多及活性氧的相关性。
将78例白细胞精子症患者精液样本和31例健康供者精液样本经Percoll梯度离心分为3层,即上层清液、30%精子层和80%精子层,采用实时荧光定量PCR法对其精子线粒体DNA含量及线粒体DNA 4977bp缺失进行定量分析,采用流式细胞术检测ROS水平。
白细胞精子症组精液中ROS水平及3层精子线粒体DNA含量均显著高于健康对照组(P<0.01)。上层清液层和80%精子层中,对照组与白细胞精子症组线粒体DNA 4977bp缺失无明显差异,但后者30%精子层显著高于对照组(P<0.01)。ROS水平与30%精子层(r=0.347,P<0.01)和80%精子层(r=0.456,P<0.01)的线粒体DNA含量呈正相关,而上层清液层无相关性。
白细胞及ROS增多可能是精子线粒体DNA含量升高的原因之一,但对线粒体DNA 4977bp缺失无明显影响。