Basic research to establish a treatment for asthenozoospermic patients.

目的：（1）研究弱精子症患者精子活性氧（ROS）的产生率及其与精子运动参数的关系。(2)材料与方法：（1）采用化学发光法测定精子活性氧（ROS）的生成量，并与正常人对照。在无刺激（稳态）或用N-甲酰基-甲硫氨酰-苯丙氨酸（f-MLP）或佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）刺激后，研究了精子制剂的ROS形成。(2)15弱精子症患者的精子，其精子产生高水平的ROS在稳态下，在体外用戊酰茶碱处理，以确定其对ROS的产生和精子运动参数的影响。(3)同样的15名患者和18名弱精子症患者， ...更多信息 用两种不同剂量的戊茶碱处理稳态时不产生ROS的精子（300毫克和1200毫克/天），以确定其对活性氧生成，精子运动参数和精子在体内的可识别性的影响。（1）当在没有刺激的情况下检测到稳态ROS形成并且其没有被f-MLP刺激时，ROS的来源可能是精子本身。(2)71例弱精子症患者中有15例的精子在稳态时产生ROS。这15例患者被分配到A组患者。(3)戊氨酰茶碱可降低A组患者精子ROS的产生，并使精子曲线运动速度和搏动交叉频率的降低保持6小时。(4)对于A组患者，口服给药喷替福林未能减少精子产生的ROS。A组患者口服低剂量（300 mg/d）戊氨酰茶碱对精子活力、运动参数和精子存活率无影响。高剂量（1200 mg/d）时，F-MLP可增加精子活力和搏动交叉频率，但对A组患者的精子存活率无影响。结论：F-MLP刺激精子制备物可确定稳态时产生ROS的来源。在弱精子症患者中，有一组患者（A组）的精子可产生可检测到的稳态ROS水平，对这一组患者，戊氨酰茶碱在体外可有效减少ROS的形成并保护精子运动参数。口服戊氨酰茶碱在低剂量下没有效果，但是，它增加了精子活力和一些精子运动参数，而不改变精子的可移植性在高dose.Future：目前，没有有效的治疗方法，如特发性弱精子症的精子功能缺陷的患者。为了开发新的治疗策略，重要的是要更多地了解这些缺陷的性质。本研究结果可能为特发性男性因素不育症的治疗提供线索。为了证实这些结果，一个前瞻性和双盲临床试验，处理生育力作为关键的结果措施，是必要的。少

Purpose : (1) To determine the incidence of reactive oxygen species (ROS) formation by spermatozoa from asthenozoospermic patients and the relationship between ROS formation and sperm motion parameters. (2) To assess the efficacy of in vitro and in vivo pentoxifylline treatment of asthenozoospermic patients whose spermatozoa generated high ROS levels.Materials and Methods : (1) ROS formation by spermatozoa from asthenozoo-spermic patients and fertile volunteers was measured by chemiluminescence. ROS formation by the sperm preparations was investigated without stimulation (steady state) or after stimulation with N-formyl-methionyleucyl-phenylalanine (f-MLP) or phorbol-12-myristate-13-acetate (PMA). (2) Spermatozoa from 15 asthenozoospermic patients, whose spermatozoa produced high levels of ROS at steady state, were treated in vitro with pentoxifylline to determine its effect on ROS generation and sperm motion parameters. (3) These same 15 patients and 18 asthenozoospermic patients whos … More e spermatozoa did not produce ROS at steady state were treated with pentoxifylline at two different dosages (300 mg and 1200 mg/day) to determine its effect on ROS generation, sperm motion parameters and sperm fertilizing ability in vivo.Results : (1) When the steady state ROS formation was detected without stimulation and that was not stimulated by f-MLP,the source of ROS could be attributed to the spermatozoa themselves. (2) Spermatozoa from 15 out of 71 asthenozoospermic patients generated ROS at steady state. These 15 patients were assigned to group A patients. (3) Pentoxifylline decreased ROS generation by spermatozoa from group A patients and preserved the decrease of curvilinear velocity and beat cross frequency for 6 hrs in vitro. (4) For group A patients orally administered pentoxifylline failed to decrease ROS generation by spermatozoa. For group A patients orally administered pentoxifylline had no effect on sperm motility, sperm motion parameters and sperm fertilizing ability at low dosage (300mg/day). It increased motility and beat cross frequency at high dosage (1200mg/day), but had no effect on sperm fertilizing ability in group A patients.Conclusions : Stimulation of sperm preparations with f-MLP can identify the source of ROS generated at steady state. Among asthenozoospermic patients, there was a group of patients (group A) whose spermatozoa produces detectable steady state levels of ROS.For this group, pentoxifylline appeared to be effective for reducing ROS formation and preserving sperm motion parameters in vitro. Orally administered pentoxifylline had no effect at low dosage, however, it increased sperm motility and some sperm motion parameters without altering sperm fertilizing ability at high dosage.Future : Currently, there is no effective treatment for patients with deficient sperm functions, such as idiopathic asthenozoospermia. To develop new treatment strategies, it is important to learn more about the nature of these deficiencies. The present results may provide clues to a therapeutic approach for the treatment of idiopathic male-factor infertility. For the confirmation of these results, a prospective and double-blind clinical trial, dealing with fecundity as the crucial outcome measure, is necessary. Less