肿瘤坏死因子α-308多态性与弱精子症关系的研究

Tumor necrosis factor α-308 polymorphism and asthenospermia

目的:探讨肿瘤坏死因子α(TNFα)基因启动子区-308位点基因型多态性与弱精子症的关系。方法:采用等位基因特异多聚酶链式反应(ASPCR)和聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)法分别对187例男性不育患者(分3组:A组,60例,为弱精子症组;B组,65例,为少弱精子症组;C组,62例,为正常精液不育组)的TNFα基因-308位点基因型进行分析。采用放射免疫分析法测定患者精浆中的TNF-α水平,并对各组数据进行统计学分析。结果:观察各组TNFα-308位点基因型GA/AA频率,与C组(8.06%)相比,A组(21.67%)和B组(26.15%)显著增高,差异均有统计学意义(P<0.05)。采用Spearman方法分析,各组间TNFα-308位点等位基因GA+AA类型与a+b级精子百分率呈显著负相关关系(r=-0.690,P<0.05)。观察各组精浆TNF-α水平,与C组[(4.03±0.66)ng/ml]相比,A组[(4.23±0.45)ng/ml]和B组[(4.29±0.47)ng/ml]显著升高,差异均有统计学意义(P<0.05)。而A组与B组相比,差异无统计学意义(P>0.05)。观察TNFα-308位点不同等位基因(GA+AA、GG)间的精浆TNFα水平,与GG类型组[(4.06±0.45)ng/ml]相比,GA+AA类型组[(4.61±0.29)ng/ml]显著升高,差异有统计学意义(P<0.05)。结论:不论精子密度如何,TNFα-308GA/AA的频率与a+b级精子百分率呈显著负相关关系,其机制之一可能与精浆TNF-α水平有关。据此推测,治疗弱精子症,抗TNFα方案可能有效;另外,精浆TNFα水平的检测可作为男性不育症的辅助诊断指标。

Objective：To investigate the relationship between-308 genotype polymorphism in the promoter region of the tumor necrosis factor alpha（TNFα）gene and asthenospermia in infertile men.Methods：Allele-specific polymerase chain reaction（ASPCR）and polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）were used to analyze the genotype at position-308 in the promoter region of the TNFα gene in 187 infertile male patients,who were divided into Groups A（asthenospermia,n = 60）,B（oligoasthenozoospermia,n = 65）and C（infertile patients with normal sperm,n = 62）.The levels of TNFα in the seminal plasma from these patients were measured by radioimmunoassay,and all the data were statistically analyzed by SPSS16.0.Results：Groups A and B exhibited significant differences from C in the frequency of GA/AA at position 308 in the promoter region of the TNFα gene（21.67% and 26.15 % versus 8.06%,P 0.05）.Spearman analysis showed a negative correlation between the GA ＋ AA type of the TNFα-308 allele and the percentage of grade a ＋ b sperm（r =-0.690,P 0.05）.The level of TNFα in the seminal plasma was significantly elevated in Groups A（[4.23 ± 0.45] ng/ml）and B（[4.29 ± 0.47] ng/ml）as compared with C（[4.03 ± 0.66] ng/ml,P 0.05）,but with no significant differences between Groups A and B（P 0.05）.It was also significantly higher in the GA＋AA（[4.61 ± 0.29] ng/ml）than in the GG type（[4.06 ± 0.45] ng/ml,P 0.05）.Conclusion：Regardless of sperm density,the frequentcy of TNFα-308 GA/AA is negatively correlated with the percentage of grade a ＋ b sperm,which may be associated with the level of TNFα in the seminal plasma.Accordingly,anti-TNFα therapy might be effective for asthenospermia,and the measurement of the TNFα level in the seminal plasma can be an auxiliary diagnostic marker for male infertility.