中英文翻译对照

PARAOXONASE-1 ACTIVITY IN SUBFERTILE （低生育力）MEN AND RELATIONSHIP TO SPERM PARAMETERS2008

对氧磷酶1活性在低生育力男性及精子参数的关系

该文提到特发性不育男性与精液异常低生育男性比较，前者显示高ROS水平，PON-1活性改变无统计学意义，后者PON-1活性降低有统计学意义

Abstract

Oxidative stress has been implicated in the pathogenesis of male infertility. Paraoxonase-1 (PON-1) is a high density lipoprotein associated antioxidant enzyme that prevents oxidative modification of low density lipoprotein. Our aims of the study were to investigate i) Seminal PON-1 activity in subfertile men ii) whether it had any relationship with semen parameters. The study included 28 idiopathic subfertile, 32 subfertile male with abnormal semen parameters and 30 fertile volunteers. Seminal PON-1 activity was measured spectrophotometrically. Seminal total antioxidant status (TAS) and total oxidant status (TOS) were determined by using colorimetric methods. Oxidative stress index (OSI) was calculated as [(TOS/TAS) ×100]. TOS and OSI were significantly higher and PON-1 activity and TAS were significantly lower in subfertile men than in idiopathic subfertile men and fertile donors. PON-1 activity was also strongly correlated with sperm concentration (r=0.68, p<0.0001), motility (r=0.58, p<0.0001) and morphology (r=0.62, p<0.0001) in overall group. The receiver operating characteristic curve (ROC) analysis revealed a high diagnostic value for PON-1 activity with respect to male factor subfertility, with an area under curve of 0.95 (95% confidence interval (CI) =0.89-1.01), sensitivity = 97 % and specificity = 88%. Men with abnormal semen parameters have decreased levels of PON-1 activity in their seminal plasma. This may play an important role in the pathogenesis of male factor subfertility.

摘要

在男性不育症的发病机制，氧化应激有牵连。对氧磷酶-1（PON- 1）是一个高密度脂蛋白相关的抗氧化酶，防止低密度脂蛋白氧化修饰。我们的研究目的是调查I）精液的PON-1活性低生育力男性二）是否有任何与精液参数的关系。这项研究包括28例特发性低生育力，30例精液参数异常的低生育力男性和32例肥沃志愿者。精液PON- 1活性的测定

分光光度法。精浆总抗氧化状态（TAS）和总氧化剂状态（TOS）

利用比色法测定。氧化应激指数（OSI）的计算公式为[（TOS/ TAS）×100]。TOS和OSI显著较高的PON- 1活性和TAS显著低于低生育力男性特发性低生育力的男性和肥沃的捐助者。PON- 1活性也很强的相关性与精子浓度（R =0.68，P<0.0001），活力（R =0.58，P

<0.0001），并在集团整体形态（R =0.62，P <0.0001）。受试者工作特征曲线（ROC）分析显示，与男性因素subfertility方面的PON- 1活性高的诊断价值，与0.95（95％可信区间（CI）=0.89-1.01）曲线下面积，灵敏度= 97 ％，特异性=88％。男性精液参数异常的PON- 1活性下降的水平，在他们的开创性精浆。这可能男性因素subfertility发病中发挥的重要作用。

Materials and Methods

In this prospective study, semen samples were collected from 28 idiopathic subfertile, 32 subfertile male with abnormal semen parameters and 30 fertile volunteers who attended to male infertility clinic. The study was approved by Harran University‘s Institutional Review Board and informed consent was obtained from each participating man. A detailed medical history was obtained from all subjects, including reproductive history and infertility evaluation of the female partner. An experienced urologist performed the genital examinations. All the patients had normal female partners aged between 18-35 years, had normal reproductive history, normal ovulation (by follicular ultrasound scan, luteal phase 3progesterone levels, and endometrial biopsy), and tubal patency (hysterosalpingogram All couples presenting for infertility evaluation had a minimum of 1 year of unprotected intercourse. Male factor subfertility was defined by the presence of at least one of the sperm anomalies oligoozoo-, asthenozooand/ or teratozoospermia. Idiopathic subfertility group had normal standard semen parameters on repeated analyses, normal genital exam. The exclusion criteria were age older than 35, smoking, alcohol drinking, coronary artery disease, unstable angina, myocardial infarction, any operation or cardiovascular intervention within the previous 3 months, hypertension, hyperlipidemia, rheumatological or endocrine conditions such as diabetes, acute-chronic liver diseases, renal dysfunction, anemia, parasitic diseases, systemic or local infection, leukocytospermia , or with any history of cancer in the past 5 yr, therapeutic interventions known to influence antioxidants such as supplemental vitamins. A group of healthy subjects of proven fertility (initiated a successful pregnancy within the last 12 months before participation in the study) that were volunteered without payment served as the

control group. Hyperlipidemia was defined as follows: serum LDL cholesterol ≥160 mg/dL or total cholesterol (TC) ≥240 mg/dL or triglyceride (TG) ≥200 mg/dL or HDL cholesterol <40 mg/dL (Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults, 2001). A patient was considered as diabetic with a fasting plasma glucose level ≥126 mg/dL.

材料和方法

在这项前瞻性的研究，收集精液样本从28特发性低生育力，32

低生育力男性精液参数异常和肥沃的志愿者30人出席男性不育症诊所。这项研究是由哈兰大学的机构审查委员会批准，并从每个参与的人获得知情同意。详细的病史是获得所有科目，包括生育史，不孕不育的女性伙伴的评价。一个有经验的泌尿科进行生殖器检查。所有患者年龄在18-35岁之间且女性配偶正常，有正常生育史，正常的排卵（卵泡超声扫描，黄体期的3黄体酮水平，和子宫内膜活检），和输卵管通畅（hysterosalpingogram所有夫妇不孕的评价介婚后正常无性保护措施1年内为神韵怀孕者。男性因素低生育组定义存在至少一个精子异常oligoozoo，asthenozooand /或teratozoospermia（畸形精子症），特发性低生育组常规精液参数的反复分析，常规生殖器检查。排除标准是年龄35岁以上，吸烟，饮酒，冠心病，不稳定型心绞痛，心肌梗死，任何操作或心血管介入前3个月内，高血压，高血脂，风湿或内分泌条件，如糖尿病，急性慢性肝病肾功能不全，贫血，寄生虫病，全身或局部感染，白细胞精子症，或与任何癌症在过去5年的历史中已知的影响，如补充维生素抗氧化剂的治疗干预。发起了一个行之有效的生育健康科目组（在过去12个月前在参与成功怀孕研究），自愿不付款服务作为对照组。

高脂血症是定义如下：血清低密度脂蛋白胆固醇≥160 mg / dL或总胆

固醇（TC）≥240毫克/升或甘油三酯（TG）≥200 mg / dL或高密度脂蛋白胆固醇<40毫克/升（专家小组检测，评估和治疗，在成人，2001年），高血脂。病人视为糖尿病与空腹血糖水平≥126毫克/升。

Measurement of PON-1 activity in seminal plasma

PON-1 activity was determined by using paraoxon as a substrate and measured by increases in the absorbance at 412 nm due to the formation of 4-nitrophenol as already described (Verit et al., 2008). Briefly, the activity was measured at 25oC by adding 50 μL of seminal plasma to 1 mL Tris-HCl buffer (100 mM at pH 8.0) containing 2 mM CaCl2 and 5.5mM of paraoxon. The rate of generation of 4- nitrophenol was determined at 412 nm. Enzymatic activity was calculated by using the molar extinction coefficient 17 100 M-1 cm-1.

测量的PON- 1活性精浆

PON- 1活性测定氧磷作为基材使用，在增加测量由于形成4 - 硝基酚在412 nm处的吸光度（Verit等，2008）。简单地说，在25℃至1 mL的Tris- HCl缓冲液加入50μL精浆活性测定在pH值8.0（100毫米），其中包含2毫米氯化钙和对氧磷5.5mm的。率的4代- 在412 nm处对硝基酚被确定。酶的活性，计算使用的摩尔消光系数17 100的M - 1 cm - 1处。Results

Demographic, semen characteristics, seminal TOS, TAS and PON-1 activity in male factor subfertility, idiopathic subfertility and fertile donors were summarized in Table 1. There was no significant difference（显著性差异）in terms of age between the groups. Seminal TOS was significantlyhigher and seminal TAS and PON-1 activity were significantly lower in male factor subfertility group as compared with idiopathic infertile men and fertile donors (Table 1).5 The relationship between seminal TOS, TAS, OSI and PON-1 activity and semen parameters in overall group (n=90) were shown in Table 2. There were negative correlations between TOS and OSI and sperm parameters such as concentration, motility and morphology (p<0.0001, for all) (Table2). In addition, TAS and PON-1 activity had significant positive correlations with all sperm parameters (p<0.0001, for all) in the study (Table2). PON-1 had a positive correlation between seminal TAS (r= 0.76, p<0.0001) and were negatively correlated with seminal TOS (r= -0.61, p <0.0001) and OSI (r= -0.73, p<0.0001) in the study. ROC analysis revealed a high diagnostic value for PON-1 activity with respect to male factor subfertility, with an area under curve (AUC) of 0.95 (95% confidence interval (CI)

=0.89-1.01), sensitivity = 97 % and specificity = 88% with a cut off value of 1.75 U/L (lower than that value was related to male factor subfertility) that was shown in Figure 1. However, PON-1 activity was not effective statistically in its diagnosis of idiopathic infertility

(AUC<0.5).

结果

人口，精液特性，精液TOS，TAS和PON - 1活性在男性因素表1总结了subfertility，特发性subfertility和肥沃的捐助者。组与组之间在年龄上没有显著差异。精囊炎服务条款显着更高，开创性的TAS和PON - 1活性显着降低，与特发性不育男性和肥沃的捐助者（见表1）相比，男性因素subfertility组。5开创性的TOS，TAS，OSI和PON - 1活性和总体组的精液参数（N =90）之间的关系如表2所示。TOS和OSI 和精子浓度，活力和形态（P <0.0001）（表2），如参数之间的负相关。此外，TAS和PON- 1活性与精子参数显著的正相关关系（P

<0.0001，为所有）在研究中（表2）。PON - 1了开创性的TAS（R =0.76，P <0.0001）之间呈正相关，并具有开创性的TOS（R =-0.61，P <0.0001）和OSI（R =-0.73，P <0.0001）呈负相关研究。ROC分析显示，男性因素为PON- 1活性较高的诊断价值subfertility与曲线下面积（AUC）为0.95（95％可信区间（CI）=0.89-1.01），灵敏度=97％，特异性=1.75 U / L，与关闭值削减88％（低于该值有关男性因素subfertility），在图1所示。然而，PON- 1活性特发性不育症的诊断方面无统计学意义（AUC<0.5）。

Discussion

In this study, we found that TOS and OSI were increased and TAS and PON-1 activity were decreased in male subfertile men with abnormal semen parameters as compared with idiopathic subfertile group and fertile donors. There was a close relationship between PON-1 activity and abnormal semen parameters in the study. Moreover, ROC curve analysis revealed a good predictive power to discriminate subfertile patients from other population. ROS have been found to have dual effect on human spermatozoa. Sperm plasma membrane has a high concentration of polyunsaturated fatty acids, which can undergo lipid peroxidation initiated

by ROS (Saleh et al., 2003). Such peroxidative damage to the sperm plasma membrane leads to a loss of membrane fluidity and integrity as a result of which the spermatozoa lose their competence to participate

in the membrane fusion events associated with fertilization (Aitken and Fischer, 1994; Alvarez and

Storey,1995; Storey, 1997). In addition ROS are also known to attack DNA, inducing strand breaks and

oxidative base damage in human spermatozoa (Hughes et al., 1996; Kodoma et al., 1997).

It has been indicated that levels of ROS were negatively correlated with the quality of sperm in

the original semen (Gomez et al., 1998). High levels of ROS production in human ejaculates may

originate from morphologically abnormal spermatozoa and/or seminal leukocytes (Aitken et al, 1990).

Many studies have reported that spermatozoa from oligozoospermic or asthenozoospermic men showed a

greater production of oxidative stress (Aitken et al, 1992; Sharma and Agarwal, 1996; Griveau and de

Lannou, 1997; Pasqualotto et al., 2000). We also found that serum TOS was significantly higher in 6 subfertile men with abnormal semen parameters in this study. Moreover, TOS was negatively correlated with semen parameters such as concentration and motility and morphology in the study.

Previous reports have described that patients with idiopathic male infertility have elevated levels

of ROS (Pasqualotto et al., 2000; Saleh et al., 2003). It has been also suggested that lipid peroxidation of

sperm membrane may be one of the key mechanisms involved in the pathophysiology of idiopathic male

infertility (Alkan et al., 1997).However, the correlation between oxidative stress and male idiopathic

infertility is not clear and there were some limitations in these studies. Saleh et al. found significant

correlations between abnormal sperm parameters including leukocytospermia and oxidative stress, but did

not state the level of leukocytospermia in each group in the study population. Morever, infertile men with

normal semen parameters had increased oxidative stress than in normozoospermic fertile donors in this

study and it is a matter of debate how oxidative stress was increased in that group with normal semen

parameters. In another study that was reported by Pasqualotto et al., the study size was small and poorly

designed since the normospermic group was not homogenous that was also consisted of varicocele

patients (Pasqualotto et al., 2000). Many studies have demonstrated that oxidative stress is increased in

varicocele even in the presence of normal semen parameters (Agarwal et al., 2006; Pasqualotto et al.,

2008; Smith et al., 2006). In this study, we found that there was no difference in seminal oxidative stress

between idiopathic subfertile men and fertile donors that supported our other previous work (Verit et al.,

2006). Moreover, Ochsendorf et al. (1998) found that spermatozoa of oligozoospermic patients contained

much lower thiol glutathione, the endogenous antioxidant, concentrations than those of normozoospermic

men. Another study also demonstrated oxidative stress was increased in asthenozoospermic patients

compared with normozoospermic men (Tavilani et al., 2005). We suggest that oxidative stress is

dependent on sperm parameters but not are directly related to the diagnosis of male factor infertility.

Antioxidants are important in maintaining the oxidant-antioxidant balance in tissues. Among the

well-known biological antioxidants, superoxide dismutase, catalase, and glutathione peroxidase/reductase

system have a significant role in protecting the sperm against peroxidative damage (De Lamirande and

Gagnon, 1993; Sharma and Agarwal, 1996). Depressed seminal antioxidant capacity has been implicated

in male subfertility. TAS levels have been shown to be lower in the semen of subfertile men as compared

with fertile men (Lewis et al., 1995, 1997; Smith et al., 1996). More specifically, Raijmakers et al. (2003)

reported significantly higher seminal plasma thiol glutathione concentrations in fertile men compared

with subfertile men. In accordance with this finding, it has been reported that ascorbate levels were

significantly reduced in seminal plasma of asthenozoospermic infertiles (Lewis et al, 1997). Furthermore,

studies have suggested that infertile men empirically treated with antioxidants have demonstrated

improved semen characteristics, fertilization in vitro, and higher pregnancy rates in the treatment group

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(Lenzi et al., 1993; Geva et al., 1996). In our study, TAS was significantly decreased in subfertile men

with abnormal semen parameters, but in idiopathic infertile group in the study.

PON-1 is an antioxidant enzyme that is highly effective in preventing lipid peroxidation of LDL

(Mackness et al., 1993). It is principally responsible for the breakdown of lipid peroxides before they

accumulate on LDL (Mackness et al., 1993). PON-1 can also destroy hydrogen peroxide

(H2O2), a major

ROS produced under oxidative stress during atherogenesis (Aviram et al., 1998) and increase the LDL

clearance (Shih et al., 2000).

PON-1 also protects HDL against lipid peroxidation (Mackness et al.,1993; Aviram et al., 1998;

Rozenberg et al., 2003). Inhibition of HDL oxidation by PON-1 preserves the antiatherogenic effects of

HDL in reverse cholesterol transport (Aviram et al., 1998). The antioxidant effect of HDL is also

assumed by PON-1 (Aviram et al., 2004).

The association between PON-1 activity and male infertility is unknown. PON-1 activity was significantly lower in male subfertile patients compared with idiopathic infertile men and fertile donors in

the present study. There were also significant positive correlations between PON-1 activity and semen

parameters such as concentration, motility and morphology. We suggest that decreased

PON-1 activity

must be related to enhanced production ROS. In addition, it has been previously shown that PON-1

activity was decreased in some diseases due to ROS pathogenesis under oxidative stress and inflammation

condition such as diabetes, coronary artery disease and endometriosis (Ayub et al., 1999; Durrington et

al., 2001; Verit et al., 2008).

The most widely used methods for measuring reactive oxygen species are colorimetric, fluorescence, chemiluminescence and electron spin resonance (ESR) spectroscopy (Tarpey et al., 2004).

We measured total oxidant levels in seminal plasma by colorimetric method that was developed by Erel in

the study (Erel, 2005). This technique has many advantages. Various other methods that have been

developed for measuring TOS had no accepted reference method. In addition, this method had final

decision concerning the standardizations, the terms and the units (Erel, 2005). Moreover, the fluorescence, chemiluminescence and ESR methods need sophisticated techniques and, in most routine

clinical biochemistry laboratories, these improved systems are not available. Measurement of TAS within semen can be conducted in a variety of ways. The ability of

seminal

plasma to inhibit chemoluminescence elicited by a constant source of ROS (horse-radish peroxidase) is a

commonly used technique. The TAS is usually quantified against a Vitamin E analogue (Trolox) and

expressed as a ROS-TAS score (Sharma et al., 1999). However, colorimetry techniques based on the

color change of ABTS (2,2’- azinobis3-ethylbenzo-thiazoline-6-sulphate) are now becoming more

popular as they are cheaper and easier to perform (Said et al., 2003; Erel, 2004). The reduced ABTS

molecule is oxidized to ABTS+ using hydrogen peroxide and a peroxidase to form a relatively stable

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blue-green color measured at 600 nm with a standard spectrophotometer. Antioxidants present within

seminal plasma suppress this color change to a degree that is proportional to their concentrations. Again

the antioxidant activity is quantified using Trolox.

In conclusion, our results showed that TOS was significantly higher and TAS and PON-1 activity

were significantly lower in patients with male subfertility, but not in idiopathic infertile group. Reduced

PON-1 activity may play a role in the pathogenesis of male subfertility. Therefore both protection from

oxidative stress and increases in PON-1 activity could be used as a powerful tool for the prevention of

subfertility.

讨论

在这项研究中，我们发现，TOS和OSI增加和TAS和PON - 1活性在精液参数异常的男性低生育组下降，与特发性低生育力组和肥沃的捐助者相比。在研究中PON - 1活性和精液异常有密切的关系。此外，ROC 曲线分析显示良好的预测能力歧视从其他人口低生育力的患者。活性氧已被发现对人类精子的双重效果。精子质膜有高浓度的多不饱和脂肪酸，它可以接受脂质过氧化活性氧发起（萨利赫等，2003）。这种精子质膜过氧化损伤导致的损失作为一个结果，其中精子失去其职权参与细

胞膜的流动性和完整性在施肥（Aitken和1994年，菲舍尔的膜融合事件;阿尔瓦雷斯和层高，1995;层高，1997年）。此外ROS的也被称为攻击DNA，诱导链断裂和人类精子的氧化基础伤害（Hughes等，1996;。Kodoma等，1997）。已经指出ROS水平呈负相关，与精子质量原精液（Gomez等。，1998）。人类精液ROS产生的高层次源于形态异常的精子和/或精液白细胞（艾特肯等，1990）。许多研究报告显示，从oligozoospermic或asthenozoospermic男性精子一个提高生产的氧化应激（艾特肯等人，1992年，夏尔马和阿加瓦尔，1996年; Griveau 和deLannou，1997年; Pasqualotto等，2000）。我们还发现，血清中的服务条款是显著较高6在这项研究中的低生育力男性精液参数异常。此外，TOS呈负相关与精液参数，如浓度和活力，并在研究形态。以前的报告中所述，特发性男性不育症患者有较高水平ROS （Pasqualotto等，2000。萨利赫等，2003）。它也已建议，脂质过氧化

精子膜可在特发性男性的病理生理所涉及的主要机制之一

然而，氧化应激和男性特发性之间的相关性不孕症（Alkan等，1997）。不孕不明确，在这些研究中有一定的局限性。萨利赫等。发现显著

畸形精子参数之间的相互关系，包括白细胞精子症和氧化应激反应，但那样

没有国家的水平在各组研究人群中的白细胞精子症。Morever，与不育男性

正常精液参数normozoospermic在这肥沃的捐助者比增加氧化应激

研究和氧化应激是如何增加在该组中，正常的精液，这是一个有争议的问题

参数。在另一个Pasqualotto等人的研究报告，研究规模小，低

设计自normospermic组不是同质的，也包括精索静脉曲张

患者（Pasqualotto等，2000）。许多研究表明，在精索静脉曲张患者氧化应激增加，甚至在精液参数正常的对照组中也是增加的（阿加瓦尔等，2006年。Pasqualotto等。2008年，史密斯等人，2006年）。在这项研究中，我们发现，特发性低生育力的男性和肥沃的捐助者精液氧化应激没有区别，支持我们的其他前期工作（Verit等。2006年）。此外，Ochsendorf等。（1998年）发现oligozoospermic（少精子症）患者的精子中低得多的谷胱甘肽巯基，内源性的抗氧化剂，浓度比ormozoospermic

男子。另一项研究也表明，氧化应激增加asthenozoospermic（少弱精子症）患者

相比与normozoospermic男子（Tavilani等，2005）。我们建议，氧化应激是但不依赖于精子参数都直接关系到男性因素不孕的诊断。抗氧化剂是重要的，在维护组织中的氧化剂抗氧化剂的平衡。其中知名的生物抗氧化剂，超氧化物歧化酶，过氧化氢酶，谷胱甘肽过氧化物酶/还原酶系统具有防止过氧化损伤的精子的一个重要的角色（德Lamirande 和Gagnon说道，1993年，夏尔马和阿加瓦尔，1996年）。降低精液抗氧化能力有关连与男性低生育。TAS水平已被证明在低生育力男性精液的降低相比生育男性（刘易斯等人，1995年，1997年。Smith等，1996）。更具体地说，Raijmakers等。（2003年）报告显著较高的

生育男性精浆巯基谷胱甘肽在浓度比较低生育力男性。根据这一发现，它已被报道，抗坏血酸水平显著减少精浆asthenozoospermic受精（刘易斯等人，1997年）。此外，建议的研究已经证明，经验与抗氧化剂治疗的不育男性改善精液特性，在体外授精，在治疗组得到更高的怀孕率7（Lenzi等，1993;。Geva等，1996）。在我们的研究中，TAS低生育力男性的显着下降与精液参数异常，但在特发性不育组的研究。PON - 1是一种抗氧化剂是高度有效地防止脂质过氧化低密度脂蛋白的酶（Mackness等，1993）。它是主要负责击穿脂质过氧化物才积累低密度脂蛋白（Mackness等，1993）。PON - 1也可以摧毁双氧水（H2O2），主要活性氧下产生氧化应激在动脉粥样硬化（Aviram等，1998），并增加低密度脂蛋白间隙（施等，2000）。PON - 1还对脂质过氧化保护的高密度脂蛋白（Mackness等，1993; Aviram等人，1998年。Rozenberg等人，2003年）。PON - 1高密度脂蛋白氧化的抑制作用保留的抗动脉粥样硬化的影响高密度脂蛋白胆固醇的逆向转运（Aviram等，1998）。高密度脂蛋白的抗氧化效果也承担PON - 1（Aviram等，2004）。PON - 1活性与男性不育之间的关联是未知的。PON - 1活性在低生育力男性患者明显降低相比，特发性不育男性和肥沃的捐助者目前的研究。也有显著的正相关关系之间的PON - 1活性和精液参数，如浓度，活力和形态。我们建议下降的PON - 1活性必须加强生产活性氧有关。此外，先前已表明，PON - 1活动减少ROS的发病机制是由于在某些疾病的氧化应激和炎症条件，如糖尿病，冠状动脉疾病和子宫内膜异位症（阿尤布等人，1999年; Durrington等。人，2001

年Verit等，2008）。

测量活性氧的最广泛使用的方法是比色法，荧光，化学发光法和电子自旋共振（ESR）光谱（Tarpey等，2004）。我们Erel开发中的比色法测定精浆中总氧化剂水平的研究（Erel 2005年）。这种技术有许多优点。各种其他方法已测量服务条款有没有公认的参考方法。此外，这种方法最后关于标准化，术语和单位的决定（Erel，2005年）。此外，荧光，化学发光法和ESR方法需要先进的技术和最常规，临床生化实验室，这些改进后的系统不可用。精液内可以进行多种方式的TAS测量。精液的能力血浆抑制化学发光引起的ROS恒流源（辣根过氧化物酶）是常用的技术。助教通常是量化对维生素E类似物（Trolox的）作为ROS - TAS评分（Sharma等人，1999年）来表示。然而，比色法技术的基础上，颜色的变化ABTS（2,2' - azinobis3 - ethylbenzo - 噻唑啉- 6 -硫酸）现在越来越流行，因为它们更便宜，更容易执行（赛义德等，2003; Erel，2004年）。减少ABTS分子被氧化ABTS +使用过氧化氢和过氧化物形成一个相对稳定的8衡量一个标准的分光光度计在600 nm的蓝绿色。抗氧化剂目前内精浆抑制这种颜色的变化在一定程度上其浓度成正比。一次量化使用Trolox的抗氧化活性。总之，我们的研究结果表明，TOS显着高于TAS和PON - 1活性在与男性subfertility患者显着降低，但在特发性不育组。减少PON - 1活性在男性subfertility的发病可能发挥的作用。因此，这两个保护氧化应激和PON - 1活性增加，可作为一个强大的工具，为预防低生育力。

Paraoxonase-1 is only present in

traceable amounts in seminal fluid and does not show any relationship with male

subfertility

对氧磷酶1溯源精液量只有目前并没有显示任何与男性subfertility的关系

Immunohistochemical analyses showed high levels of PON1 and PON3 expression

in testicular tissue. PON2 expression was also detected, albeit at weaker levels. Oxidative stress indicators in biopsies were low and localized in some specific areas of the seminiferous tubules. PON1 was detected in seminal fluid at very low levels but with no significant differences between patients and controls. Receiveroperating characteristic analysis showed a low diagnostic power of semen PON1 levels

免疫组化分析表明：高层次的PON1和PON3的表达睾丸组织。PON2表达也发现，尽管在较弱的水平。活检中的氧化应激指标低，在一些具体领域的本地化曲细精管。PON1是精液中检测到非常低的水平但无显着性差异患者和对照组之间。Receiveroperating特征分析显示PON1精液的诊断能力低水平。