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1、【精品文档】如有侵权，请联系网站删除，仅供学习与交流中英文翻译对照.精品文档.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AbstractOxidative stress has been implicated in the pathogenesis of male infertility
2、. 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 st
3、udy 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. Oxidativ
4、e 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, p0.000
5、1), motility (r=0.58, p0.0001) and morphology (r=0.62, p0.0001) 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
6、.89-1.01), sensitivity = 97 % and specificity = 88%. Men with abnormal semen parameters have decreased levels of PON-1 activity in their seminalplasma. 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
7、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，P0.0001），活力（R =0.58，P 0.0001），并在集团整体形态（R =0.62，P
8、 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 MethodsIn this prospective study, semen samples were collected from 28 idiopathic subfertile, 32 subfertile mal
9、e with abnormal semen parameters and 30 fertile volunteers who attended to male infertility clinic. The study was approved by Harran Universitys Institutional Review Board and informed consent was obtained from each participating man. A detailed medical history was obtained from all subjects, includ
10、ing 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
11、 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-, astheno
12、zooand/ 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 cardi
13、ovascular 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 t
14、he 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 co
15、ntrol 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 considere
16、d 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（hysterosalpingogr
17、am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
18、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 plasmaPON-1 activity was determined by using paraoxon as a substrate and measured by increases in t
19、he absorbance at 412 nm due to the formation of 4-nitrophenol as already described (Verit et al., 2008). Briefly, the activity was measured at 25C by adding 50 L 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- nit
20、rophenol 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50L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
21、性，计算使用的摩尔消光系数17 100的M - 1 cm - 1处。ResultsDemographic, semen characteristics, seminal TOS, TAS and PON-1 activity in male factorsubfertility, 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 T
22、OS 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
23、=90) were shown in Table 2. There were negative correlations between TOS and OSI and sperm parameters such as concentration, motility and morphology (p0.0001, for all) (Table2). In addition, TAS and PON-1 activity had significant positive correlations with all sperm parameters (p0.0001, for all) in
24、the study (Table2). PON-1 had a positive correlation between seminal TAS (r= 0.76, p0.0001) and were negatively correlated with seminal TOS (r= -0.61, p 0.0001) and OSI (r= -0.73, p0.0001) in the study. ROC analysis revealed a high diagnostic value for PON-1 activity with respect to male factorsubfe
25、rtility, 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 st
26、atistically in its diagnosis of idiopathic infertility (AUC0.5).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
27、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s
28、ubfertility），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（AUC0.5）。DiscussionIn this study, we found that TOS and OSI were increased and TAS and PON-1 activity weredecreased in male subfertile men with abnormal semen parameters as compared with idiopathic subfertile group and fertile donors. There was a close
29、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 conc
30、entration 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 participatein t
31、he membrane fusion events associated with fertilization (Aitken and Fischer, 1994; Alvarez andStorey,1995; Storey, 1997). In addition ROS are also known to attack DNA, inducing strand breaks andoxidative base damage in human spermatozoa (Hughes et al., 1996; Kodoma et al., 1997).It has been indicate
32、d that levels of ROS were negatively correlated with the quality of sperm inthe original semen (Gomez et al., 1998). High levels of ROS production in human ejaculates mayoriginate from morphologically abnormal spermatozoa and/or seminal leukocytes (Aitken et al, 1990).Many studies have reported that
33、 spermatozoa from oligozoospermic or asthenozoospermic men showed agreater production of oxidative stress (Aitken et al, 1992; Sharma and Agarwal, 1996; Griveau and deLannou, 1997; Pasqualotto et al., 2000). We also found that serum TOS was significantly higher in 6 subfertile men with abnormal seme
34、n 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 levelsof ROS (Pasqualotto et al., 2000; Saleh et al.,
35、2003). It has been also suggested that lipid peroxidation ofsperm membrane may be one of the key mechanisms involved in the pathophysiology of idiopathic maleinfertility (Alkan et al., 1997).However, the correlation between oxidative stress and male idiopathicinfertility is not clear and there were
36、some limitations in these studies. Saleh et al. found significantcorrelations between abnormal sperm parameters including leukocytospermia and oxidative stress, but didnot state the level of leukocytospermia in each group in the study population. Morever, infertile men withnormal semen parameters ha
37、d increased oxidative stress than in normozoospermic fertile donors in thisstudy and it is a matter of debate how oxidative stress was increased in that group with normal semenparameters. In another study that was reported by Pasqualotto et al., the study size was small and poorlydesigned since the
38、normospermic group was not homogenous that was also consisted of varicocelepatients (Pasqualotto et al., 2000). Many studies have demonstrated that oxidative stress is increased invaricocele even in the presence of normal semen parameters (Agarwal et al., 2006; Pasqualotto et al.,2008; Smith et al.,
39、 2006). In this study, we found that there was no difference in seminal oxidative stressbetween 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 containedmuch
40、 lower thiol glutathione, the endogenous antioxidant, concentrations than those of normozoospermicmen. Another study also demonstrated oxidative stress was increased in asthenozoospermic patientscompared with normozoospermic men (Tavilani et al., 2005). We suggest that oxidative stress isdependent o
41、n 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 thewell-known biological antioxidants, superoxide dismutase, catalase, and glutathione peroxidase/reductasesystem ha
42、ve a significant role in protecting the sperm against peroxidative damage (De Lamirande andGagnon, 1993; Sharma and Agarwal, 1996). Depressed seminal antioxidant capacity has been implicatedin male subfertility. TAS levels have been shown to be lower in the semen of subfertile men as comparedwith fe
43、rtile 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 comparedwith subfertile men. In accordance with this finding, it has been reported that ascorbate levels w
44、eresignificantly reduced in seminal plasma of asthenozoospermic infertiles (Lewis et al, 1997). Furthermore,studies have suggested that infertile men empirically treated with antioxidants have demonstratedimproved semen characteristics, fertilization in vitro, and higher pregnancy rates in the treat
45、ment group7(Lenzi et al., 1993; Geva et al., 1996). In our study, TAS was significantly decreased in subfertile menwith 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 e
46、t al., 1993). It is principally responsible for the breakdown of lipid peroxides before theyaccumulate on LDL (Mackness et al., 1993). PON-1 can also destroy hydrogen peroxide (H2O2), a majorROS produced under oxidative stress during atherogenesis (Aviram et al., 1998) and increase the LDLclearance
47、(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 ofHDL in reverse cholesterol transport (Aviram et al., 1998). The antioxidant effect o
48、f HDL is alsoassumed by PON-1 (Aviram et al., 2004).The association between PON-1 activity and male infertility is unknown. PON-1 activity wassignificantly lower in male subfertile patients compared with idiopathic infertile men and fertile donors inthe present study. There were also significant pos
49、itive correlations between PON-1 activity and semenparameters such as concentration, motility and morphology. We suggest that decreased PON-1 activitymust be related to enhanced production ROS. In addition, it has been previously shown that PON-1activity was decreased in some diseases due to ROS pathogenesis under oxidative stress and inflammationcond