论文部分内容阅读
目的:分析特发性男性不育患者催产素(oxytocin,OT)水平及其受体的表达。方法:选取特发性少精子症(oligozoospermia group,OG)(含无精子)、弱精子症(asthenozoospermia group,AG)及少精子合并弱精子症(oligoasthenozoospermia group,OAG)男性不育患者共65例,年龄20～45岁;对照组(control group,CG)选取有自然生育史的健康男性志愿者20例(精液参数正常),年龄20～45岁。检测各组血清黄体生成素(luteinizing hormone,LH)、卵泡刺激素(follicle-stimulating hormone,FSH)、睾酮(testosterone,T)及OT含量,进一步通过检测催产素受体启动子功能序列(oxytocin receptor promotor fuctional sequence,OTRP)、催产素受体羧基端部分mRNA序列(OTRmR-NA)以及OTR蛋白构象、组成来分析OTR表达情况。OTR蛋白印迹分析结果经灰度软件处理后转化为计数资料;统计方法采用单因素方差分析。结果:①男性生殖内分泌激素含量分析:OT:各病例组均高于CG[(79.30±3.83)pg/ml],具有统计学意义(F0.05/2(2,82)=8.29,P<0.001);LH:AG[(4.26±0.31)IU/L]及OAG[(4.55±0.40)IU/L]均低于OG[(6.77±0.57)IU/L]和CG[(7.19±0.50)IU/L](F0.05/2(2,82)=11.64,P<0.01);FSH:AG[(5.02±0.39)IU/L]低于CG[(8.91±0.91)IU/L]、OG[(11.86±1.76)IU/L]及OAG[(8.82±1.03)IU/L](F0.05/2(2,82)=7.22,P<0.01);T:各组间无统计学差异(F0.05/2(2,82)=0.42,P=0.739)。②OTR基因转录分析:OTR启动子功能序列和OTR成熟mRNA序列未发现明显的变异。③OTR构象分析:人OTR以单体和寡聚体同时存在,以寡聚体为主,常见为四聚体和六聚体;AG(0.41±0.03)和OAG(0.13±0.01)的单体明显多于OG(0.05±0.004)和CG(0.05±0.003)(F0.05/2(2,82)=115.50,P<0.01);AG中20%病例存在寡聚体明显减少现象。结论:男性不育患者血清OT含量及OTR表达的差异提示OT与男性不育间存在一定的联系,OTR单体表达升高及寡聚体表达降低为进一步探索OT与男性不育间的关系提供了新的方向。
Objective: To analyze the level of oxytocin (OT) and its receptor expression in idiopathic male infertility. Methods: A total of 65 male infertile patients with oligozoospermia group (OG) (including azoospermia), asthenozoospermia group (AG) and oligoasthenozoospermia group (OAG) , 20 to 45 years old; control group (control group, CG) selected natural male reproductive history of 20 healthy volunteers (sperm parameters were normal), aged 20 to 45 years. The levels of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T) and OT in serum of each group were detected. The levels of oxytocin receptor promotor fuctional sequence (OTRP), OTRmR-NA of oxytocin receptor and OTR protein conformation and composition to analyze OTR expression. The result of OTR Western blot analysis was converted into count data by gray scale software. Statistical methods were analyzed by one-way ANOVA. Results: (1) Male reproductive endocrine hormone content analysis: OT: Each case group was higher than CG [(79.30 ± 3.83) pg / ml], with statistical significance (F0.05 / 2 (2,82) = 8.29, P < 0.001); LH: AG [4.26 ± 0.31] IU / L and OAG 4.55 ± 0.40 IU / L were lower than those of OG [6.77 ± 0.57] IU / L and CG [(7.19 ± 0.50) (5.02 ± 0.39) IU / L] was lower than that of CG [(8.91 ± 0.91) IU / L] OG [(11.86 ± 1.76) IU / L] and OAG [(8.82 ± 1.03) IU / L] (F0.05 / 2 Difference (F 0.05 / 2 (2,82) = 0.42, P = 0.739). ② OTR gene transcription analysis: OTR promoter function sequence and OTR mature mRNA sequence showed no significant variation. OTR conformational analysis: human OTR co-exist in the monomer and oligomers, mainly oligomers, tetramer and hexamer common; AG (0.41 ± 0.03) and OAG (0.13 ± 0.01) monomer was significantly More than OG (0.05 ± 0.004) and CG (0.05 ± 0.003) (F0.05 / 2 (2,82) = 115.50, P <0.01). There was a significant decrease of oligomers in 20% of AG cases. CONCLUSIONS: The difference of serum OT content and OTR expression in male infertility suggests that there is a certain relationship between OT and male infertility, elevated expression of OTR monomer and low expression of oligomer in order to further explore the relationship between OT and male infertility A new direction