rhTNF-α对人精子凋亡率和膜表面肿瘤坏死因子受体Ⅱ表达量影响的实验研究

Experimental Study on the Impact of rhTNF - α in Vitro on Human Sperm Apoptosis Rate and the Expression of TNFR - Ⅱ on the Surface of Sperm Cell Membrane

目的：研究重组人肿瘤坏死因子-α（rhTNF-α）在体外对人精子凋亡率和精子膜表面肿瘤坏死因子受体Ⅱ（TNFR-Ⅱ）表达量的影响，探讨rhTNF—α在体外对人精子质量影响的作用机制。方法：26例检验合格精液标本经Percoll液洗涤处理，Ham，sF10液将精子密度调整为10x10。／ml，每份标本各取3m1分装于两支无菌试管（1．5ml／管），分为空白对照组和实验对照组两组，每组26份标本。空白对照组试管内不加rhTNF—α，实验对照组加入终浓度为270μg／ml的rhTNF-α，置37℃、5％CO2培养箱孵育2h，然后用流式细胞仪分别检测两组的精子凋亡率和精子膜表面TNFR-Ⅱ表达量，每份标本均检测10000个精子，比较两组精子凋亡率和TNFR-Ⅱ表达量的差异。结果：实验对照组与空白对照组人精子凋亡率分别为8．28％和5．07％，人精子膜表面TNFR-Ⅱ表达率分别为52．09％和73．79％，差异均有统计学意义（P〈0．01）。结论：rhTNF-α体外作用后使精子凋亡率明显升高，膜表面TNFR-Ⅱ的表达量明显降低，TNF-α与精子膜表面受体结合后可能促使精子凋亡而影响精子的质量。

Objective ： To investigate the impact of recombinant human tumor necrosis factor - α （ rhTNF - α） in vitro on human sperm apoptosis rate and the expression of tumor necrosis factor receptor - Ⅱ （ TNFR - Ⅱ ） on the sperm membrane surface and its regulatory mechanisms. Methods： 26 cases qualified sperm samples were treated by Percoll liquid, and the sperm density was adjusted to 10 × 10^6/ml with Ham, s FIO liquid. Then each sample was taken out 3 ml and subpackaged in two sterile tubes （ 1.5 ml per tube） , a total of 52 samples were divided into control group and experimental group each group contains 26 samples. The samples in control group were not treated by rhTNF - α while which in the experimental group were treated with rhTNF - α （270 μg/ml）. Two hours after incubated （37℃, 5% CO2 incubator）, all samples were detected by flow cytometry, each specimen was tested 10,000 sperms. The sperm apoptosis rate and the expression of tumor necrosis factor receptor - Ⅱ （ TNFR -Ⅱ ） on the sperm membrane surface between the two groups were compared respectively. Results： The human sperm apoptosis rates were 8.28% and 5.07% （P 〈0.01 ） in experimental group and control group respectively, and the expression rates of tumor necrosis factor receptor- Ⅱ on the sperm membrane surface were 52.09 % and 73.79 % （P 〈 0. 01 ） respectively, there was a significant difference between the two groups. Conclusions： The human sperm apoptosis rate was higher and the expression of TNFR - Ⅱ on the sperm membrane surface was significantly lower after treated by rhTNF - α in vitro. TNF - α combined with tumor necrosis factor receptor- Ⅱ on the sperm membrane may play an important role in the regulation of sperm apoptosis.