精索静脉曲张不育患者氧化应激水平和精子DNA完整性及精液参数的相关性分析

目的:分析精索静脉曲张(VC)不育患者氧化应激同DNA完整性及精液参数的关系。方法:采用前瞻性研究,纳入98例患者,根据活性氧水平(ROS)将VC不育患者分为ROS低水平组(54例)及高水平组(44例),分析两组精子DNA碎片指数(DFI)、精子活力、形态等参数的差异,并分析它们之间的相关性。结果:ROS高水平组DFI显著高于ROS低水平组[(34.49±6.05)%vs(27.38±8.10)%,P=0.039]。与ROS低水平组比较,ROS高水平组精子活力[(25.21±18.22)%vs(36.16±22.83)%,P=0.017]、前向运动精子百分率[(16.34±9.22)%vs(23.34±11.53)%,P=0.041]、曲线速率[(20.62±4.38)%vs(27.03±6.21)%,P=0.013]及直线性率[(18.30±7.93)%vs(29.75±8.24)%,P=0.024]均显著降低,而精液白细胞数显著升高[(0.86±0.41)×106/ml vs(0.65±0.15)×106/ml,P=0.022]。Spearman相关性分析表明ROS水平同精液白细胞数(r=0.41,P0.01)及DFI(r=0.21,P=0.006)呈显著正相关,同曲线速率(r=-0.24,P=0.017)、直线性率(r=-0.24,P=0.021)、精子活力(r=-0.31,P=0.002)及前向运动精子百分率(r=-0.41,P=0.012)呈显著负相关。DFI同精子活力(r=-0.29,P0.01)、前向运动精子百分率(r=-0.34,P0.01)呈显著负相关。结论:精浆ROS水平同VC不育患者DFI显著正相关。氧化应激及DNA完整性可影响患者的精子参数。     

精索静脉高位结扎术对精子DNA碎片的影响

目的:探讨精索静脉高位结扎术对精子DNA碎片影响及手术效果的评价。方法:精索静脉曲张(Varicocele,VC)患者34例,均行精索静脉高位结扎术。术前和术后3个月按WHO推荐方法进行精子运动指标(10项)、精子巴氏染色形态分析和精子DNA碎片分析检测。结果:VC患者术后正常精子形态比例明显高于术前,有显著性差异(P<0.01),术后精子DNA碎片比率、精子畸形指数(Sperm deformity index,SDI)和多种异常指数(Multiple anomalies index,MAI)均明显低于术前,有显著性差异(P<0.01);术后精子浓度、b级精子率均较术前升高,有显著性差异(P<0.05);术后a级精子比率、(a+b)级精子比率均比术前明显升高,有显著性差异(P<0.01)。VCⅠ级、Ⅱ级、Ⅲ级组术后精子DNA碎片比率均较术前明显降低,差异有统计学意义(P<0.01);亚临床型VC组术后精子DNA碎片比率与术前无显著差异(P>0.05)。VC术后大晕环精子比率较术前明显增多,有显著性差异(P<0.01);术后中晕环、小晕环、无晕环精子比率均较术前降低,有显著性差异(P<0.01)。结论:行精索静脉高位结扎术能有效改善VC患者的精子质量。     

通精灵对实验性精索静脉曲张大鼠生精细胞凋亡及Bcl-2表达的影响

目的：探讨实验性精索静脉曲张大鼠睾丸生精细胞凋亡状况及通精灵对精索静脉曲张大鼠睾丸生精功能的保护作用。方法：成年清洁级雄性Wistar大鼠50只,按Sapyol法制作左精索静脉曲张大鼠模型,随机分为假手术组、模型组、通精灵低剂量组、通精灵中剂量组、通精灵高剂量组各10只。造模1个月后,各组予以不同药物灌胃1次／d。2个月后处死,用原位末端脱氧核苷酸转移酶标记法检测睾丸生精细胞凋亡,免疫组化法检测睾丸组织Bcl-2表达。结果：模型组大鼠左睾丸凋亡指数明显高于假手术组及通精灵各组（P〈0．01）,通精灵各组左睾丸凋亡指数显著低于模型组（P〈0．05）。与假手术组比较,模型组大鼠睾丸组织Bcl-2蛋白表达下降（P〈0．05）,与模型组比较,通精灵中、高剂量组睾丸组织Bcl-2蛋白表达显著上升（P〈0．05）。结论：通精灵能降低实验性精索静脉曲张大鼠睾丸生精细胞凋亡,减轻实验性精索静脉曲张大鼠睾丸组织的病理损害,上调凋亡抑制基因Bcl-2表达。     

精索静脉曲张对孕前男性精浆生化及精子DNA完整性的影响

目的 探讨精索静脉曲张(varicocele,VC)对孕前男性精浆生化及精子DNA完整性的影响.方法 孕前男性246例按照有无VC分为VC和对照组,分别检测精浆生化指标、精子DNA完整性.比较VC组与对照组精浆生化及精子DNA碎片率(DNA fragmentation index,DFI),分析VC组不同曲张程度患者精...     

精子染色质扩散实验检测精索静脉曲张及不明原因不育患者精子DNA碎片

目的了解精索静脉曲张(VC)及不明原因不育患者精子DNA碎片的发生比例。方法改进的精子染色质扩散(SCD)实验分析精子DNA碎片。检测VC不育患者39例,不明原因不育患者57例。以生育健康成年男性32例为对照组。结果VC不育患者SCD小光晕和无光晕精子(精子DNA碎片)比值平均为(36.6±18.9)%,VC不育组明显高于对照组(12.1±5.2)%(P<0.001),而大光晕和中光晕精子比值VC不育组明显低于对照组(P<0.01);不明原因不育患者精子DNA碎片比值平均为(26.8±10.2)%,与对照组[(12.1±5.2)%]比较有显著性差异(P<0.001)。结论SCD实验表明,VC及不明原因不育患者精子DNA碎片比值增高。     

精索静脉曲张对精子染色质结构及运动能力的影响

目的:探讨精索静脉曲张对精子染色质结构及运动能力的影响。方法:对精索静脉曲张组(无慢性疾病及生殖系统病史者,n=74)和对照组(n=89)进行精子染色质结构分析(SCSA)和精液常规分析(精液量、粘稠度、pH值、精子顶体完整率、精子存活率、畸形率、精子密度、活动率及各运动参数)。结果:精索静脉曲张组精子密度、a+b级精子百分率和精子存活率[分别为(41.4±38.7)×106/ml,(31.7±16.9)%,(62.8±22.2)%]显著低于对照组[分别为(80.9±63.1)×106/ml,(46.8±20.5)%,(77.2±17.5)%],P均<0.05;精子运动参数VCL、VSL、VAP精索静脉曲张组分别为(37.4±12.5)、(23.4±7.8)、(26.5±8.2)μm/s,对照组分别为(42.4±10.7)、(27.3±7.3)、(30.7±7.8)μm/s,精索静脉曲张组与对照组相比显著降低(P均<0.05),而MAD显著增加(P<0.01),SCSACOMPαt值精索静脉曲张组(为23.2±16.2)明显高于对照组(为14.1±11.8)(P<0.05)。结论:精索静脉曲张引起精子DNA损伤和精子运动能力的改变,可能是导致男性不育的主要原因之一。     

实验性精索静脉曲张对大鼠睾丸生精细胞凋亡的影响

目的 :探讨大鼠精索静脉曲张对睾丸生精细胞凋亡的影响 ,阐明精索静脉曲张引起不育的病理机制。　方法 :选用雄性SD大鼠 32只 ,随机分为假手术组、4 5d模型组、6 0d模型组及 90d模型组。采用肾静脉缩窄法建立精索静脉曲张动物模型 ,流式细胞术分别检测各组大鼠睾丸凋亡细胞数及各级生精细胞数。　结果 :假手术组未出现明显的凋亡峰 ,各模型组均出现明显的凋亡峰 ,且随着造模时间的延长凋亡峰增高。　结论 :精索静脉曲张可引起睾丸生精细胞大量凋亡 ,各级生精细胞数减少 ,这可能是精索静脉曲张引起睾丸生精功能障碍从而导致不育的根本机制     

精索静脉曲张不育患者精子质量及其超微结构的研究

目的:研究精索静脉曲张不育患者精液质量及精子超微结构的变化。方法:不育伴精索静脉曲张患者118例作为实验组(VC组),正常自愿捐精者76例作为对照组,对其精液常规、精浆生化及外周血内分泌水平进行检测,并应用扫描和透射电镜技术对精子的超微结构进行观察。结果:VC组精液常规检查中,精子浓度、前向运动能力、存活率显著低于正常组(P<0.05),精液量、非前向运动能力无差异(P>0.05),精浆生化各项指标中,果糖浓度无显著差异,中性α-葡糖苷酶、锌离子浓度均显著低于正常组(P<0.05),外周血FSH、LH、T、E2水平均无统计学差异(P>0.05)。扫描电镜检查示正常形态精子比率低于对照组[(56.76±15.32)%vs(12.34±6.58)%,P<0.05],精子异常主要发生在头颈部,透射电镜检查示精子畸变以尖头为主,且伴有分化异常的复杂畸形。结论:VC可导致少-弱-畸精子症,进而引起不育,其原因可能是由于精浆微环境及超微结构的改变。     

一氧化氮对精索静脉曲张患者精子功能的影响

精索静脉曲张（ＶＣ）是男性泌尿生殖系统中最常见的疾病之一,其对男性生育能力的影响越来越受到重视。为了探讨一氧化氮（ＮＯ）对精索静脉曲张患者精子功能的影响。我们测定了１５例ＶＣ不育患者及１２例有生育能力的正常人的精索静脉血清、外周静脉血清和精浆中ＮＯ含量及ＮＯＳ活性,现报道如下。一、材料和方法１．一般资料　（１）实验组：１５例精索静脉曲张不育患者,年龄２２～３６岁（平均２６岁）,婚后２～１０年内不育,查体１２例左侧精索静脉曲张（Ⅰ～Ⅲ度）,３例双侧精索静脉曲张（Ⅰ～Ⅱ度）,连续３次精液分析提示弱精…     

精索静脉曲张与精子形态的相关性研究

精索静脉曲张(varcocele,VC)是男性常见的导致生殖障碍的疾病,多见于青壮年,一般以左侧为主,在男性不育症中占21%～41%~[1],其影响生育功能是多方面的.我们对183例VC手术前后精子形态进行计算机辅助精液全自动分析,了解其精子形态改变,探讨VC对精子形态的影响,分析其影响生育的具体机制,现报道如下.     

Capsaicin improves sperm quality in rats with experimental varicocele

Capsaicin is the main capsaicinoid in chilli peppers that have numerous biological and pharmaceutical roles in the body such as antioxidant, anti-inflammatory, anticarcinogenic, analgesic, counterirritant and antiarthritic properties. Numerous studies have shown increased oxidative stress in men with varicocele that is caused by dilation of the spermatic vein and increase of testicular temperature. Therefore, we aimed to assess the effect of Capsaicin on sperm parameters in rats with experimental varicocele. At first, we induced varicocele in 30 Wistar rats and, verify varicocele model only in 10 rats by assessment of sperm parameters, oxidative stress, DNA damage and persistent histone after 2 months. Of the remaining 20 varicocelised rats, half of them were treated with 2.5 mg/kg Capsaicin for two months and the other half served as control. Then, sperm tests were assessed, and the results showed that Capsaicin can restore the mean of sperm oxidative stress (38.78 ± 3.75 versus 58.37 ± 4.34; p < .05), sperm concentration (60.14 ± 7.66 versus 34.87 ± 5.78; p < .05) and motility (62.43 ± 3.10 versus 41.22 ± 5.11; p < .05) in varicocelised rats treated with Capsaicin compared to varicocelised rats that were not treat. Therefore, Capsaicin possibly with reduction of oxidative stress level could improve mean of sperm concentration and motility in varicocele condition.     

实验性精索静脉曲张对大鼠前列腺氧化应激作用的影响

目的探讨实验性精索静脉曲张(VC)对大鼠前列腺氧化应激作用的影响。方法雄性Wistar大鼠24只建立左侧VC模型作为实验组,18只行假手术作为对照组,手术后4、8和16周取前列腺,测定前列腺组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)活性及丙二醛(MDA)含量,RT-PCR法检测CuZn-SOD及GPx mRNA表达。结果手术后4周,实验组SOD、GPx活性、MDA含量及GPx mRNA水平与对照组比较差异均无统计学意义(P>0.05),而CuZn-SODmR-NA水平实验组(0.366±0.085)显著高于对照组(0.221±0.043)(P<0.05);手术后8周,SOD、GPx活性(25.61±4.06、7.07±1.35)在实验组均显著低于对照组(31.74±3.82、8.74±1.07)(P<0.05),而CuZn-SOD mRNA、GPx mRNA水平及MDA含量均显著高于对照组(P<0.05);手术后16周,SOD、GPx活性实验组(23.88±3.79、6.53±1.12)均显著低于对照组(39.46±4.39、9.05±1.34),MDA含量实验组(4.26±1.61)显著高于对照组(2.35±0.85)(P<0.05),CuZn-SOD mRNA、GPx mRNA水平差异无统计学意义(P>0.05)。结论实验性VC引起大鼠前列腺活性氧水平升高及抗氧化酶活性降低,可能是VC导致不育的原因之一。     

Testicular oxidative stress. Effects of experimental varicocele in adolescent rats

OBJECTIVE: This present study was undertaken to determine the levels of malondialdehyde (MDA), nitric oxide (NO) and total antioxidant status (TAS) in testes of adolescent rats with experimental bilateral varicocele and to determine the effects of oxidative stress on testis produced by varicocele. METHODS: 6-week-old, male Wistar rats, weighing 146-334 g (228.37 +/- 41.34 g), were randomly allocated into two groups. The first group underwent selective and bilateral partial ligation of the spermatic vein (n = 28), and the second group underwent a sham operation and served as the controls (n = 15). Animals were sacrificed 12 weeks after surgery and dilatation of the spermatic veins was observed in the first study group. Bilateral orchiectomy was performed in all rats, and MDA, NO and TAS levels were measured. RESULTS: In the study group, the mean MDA (SEM) level was 15.58 +/- 6.07 micromol/g protein, and in the control group, it was 11.59 +/- 3.86 micromol/g protein, respectively; this difference was statistically significant (p < 0.05). The mean NO level was 82.73 +/- 77.84 nmol/g protein in the study group, whereas 28.65 +/- 20.18 nmol/g protein in the control group, this difference was also statistically significant (p < 0.005). The mean TAS levels of the study and control groups were 0.91 +/- 0.32 and 1.78 +/- 0.46 nmol/g tissue, respectively, and this difference was also statistically significant (p < 0.001). But there was no correlation between these three parameters (MDA<-->TAS: r = -0.103, p > 0.05; MDA<-->NO: r = -0.104, p > 0.05; NO<-->TAS: r = -0.123, p > 0.05). CONCLUSION: These findings suggest that varicocele may change the testicular oxidative status and may play a role in testicular dysfunction that causes infertility.     

Effect of experimental varicocele in rats on testicular oxidative stress status

The present experiments were undertaken to determine the levels of MDA, SOD and catalase in the testis of adolescent rats with experimental left varicoceles. Male Wistar rats, 7 weeks old and weighing 160-170 g, were randomly allocated into three groups. The first group of rats underwent partial ligation of the left renal vein (n = 15). The second group of rats underwent a sham operation (n = 7) and the third group acted as controls (n = 7). Animals were sacrificed 6 weeks after surgery and dilatation of the internal spermatic veins was observed. Levels of MDA, SOD and catalase activity were measured in testis. The experimental left varicocele group showed severe testicular changes compared to other groups. The mean MDA (SEM) levels in right and left testicular tissues of varicocele bearing rats, sham-operated rats, and control rats were 0.48 +/- 0.24 and 0.31 +/- 0.11, 0.22 +/- 0.02 and 0.35 +/- 0.12, 0.62 +/- 0.29 and 0.13 +/- 0.05, respectively (P > 0.05). The mean SOD (SEM) levels in right and left testicular tissues of varicocele bearing rats, sham-operated rats, and control rats were 7,790 +/- 606 and 6,974 +/- 574, 7,475 +/- 1,517 and 7020 +/- 1,106, 8,727 +/- 1,188 and 9,019 +/- 1,129, respectively (P > 0.05). The mean catalase (SEM) levels in right and left testicular tissues of varicocele bearing rats,sham-operated rats, and control rats were 75.77 +/- 11.5 and 53.82 +/- 10.1, 91.94 +/- 14 and 94.90 +/- 32, 65.40 +/- 5.7 and 90.93 +/- 16.4, respectively (P > 0.05). Our results suggest that oxidative status, which reflects a relative balance between reactive oxygen species (ROS) generated and ROS scavenged, may not be responsible for the testicular dysfunction associated with experimentally induced varicocele during adolescence in rats.     

精索静脉曲张对精子DNA完整率的影响

目的 观察不同程度及不同中医证型之间精索静脉曲张患者的精子DNA完整率.方法 试验前取生育前检查正常精液标本20例,测定精子DNA完整率做预实验,其数值拟做参考指标.将78例精索静脉曲张伴不育患者根据临床症状分为轻度、中度、重度.并根据其中医证型分为瘀血阻络组、肝肾亏虚组,湿热瘀阻组,然后,测定其精子DNA完整率,分别与正常精子DNA完整率做对比.结果 轻度、中度、重度精索静脉曲张组病人的精子DNA完整性与精液常规正常组比较有显著性差异(P<0.05);不同中医证型精索静脉曲张组间病人的精子DNA完整性差异不大(P>0.05).结论 精索静脉曲张能够造成精子DNA完整性的损伤,其损伤程度与精索静脉曲张严重程度有密切关系,与中医证型关系不明显.     

Effect of varicocele on sperm function and semen oxidative stress

Study Type – Aetiology (case control) Level of Evidence 3b What’s known on the subject? and What does the study add? Varicocele leads to alterations in sperm DNA integrity even when alterations in semen quality are not yet observed in adolescents. In adults, alterations to sperm DNA are associated to altered sperm morphology, indicating that altered spermatogenesis may be an important cause for the increased sperm DNA fragmentation observed in these men. One other important cause of increased DNA fragmentation is oxidative stress, and we wished to verify if this was the case. The study adds the information that, in the adult varicocele, it is most likely that an altered testicular environment is leading to increased DNA fragmentation and decreased mitochondrial activity and acrosome integrity, because no increase in oxidative stress was observed.

OBJECTIVE

? To assess the effect of varicocele on sperm DNA integrity, mitochondrial activity, lipid peroxidation and acrosome integrity.

PATIENTS AND METHODS

? In all, 30 patients with a clinically diagnosed varicocele of grade II or III and 32 men without a varicocele were evaluated for sperm DNA fragmentation (comet assay), mitochondrial activity (3,3′‐diaminobenzidine assay), lipid peroxidation (malondialdehyde) and acrosome integrity (fluorescent probe labelled peanut agglutinin).

RESULTS

? The varicocele group showed fewer spermatozoa with intact DNA (grade II, P= 0.040), more cells with inactive mitochondria (class III, P= 0.001), fewer cells with active mitochondria (class I, P= 0.005) and fewer spermatozoa with intact acrosomes (P < 0.001). Finally, no significant differences were observed in lipid peroxidation levels.

CONCLUSION

? Men with varicocele showed an increase in sperm DNA fragmentation and a reduction in mitochondrial activity and acrosome integrity. However, lipid peroxidation levels remained unchanged.     

薯蓣多糖体外对精子存活率和DNA完整性的影响

目的:通过观察薯蓣多糖体外对精子膜及DNA完整性的影响。为少弱精子症的治疗提供新的途径。方法:正常已生育成年男性,禁欲3~5 d手淫法取精液。分为6组,每组各取0.2 ml精液;对照组为精液原液,其他5组分别加入等量的生理盐水、维生素C溶液及不同浓度薯蓣多糖溶液(0.25、1.0、5.0 mg/ml)。通过伊红-Y染色试验、精子低渗肿胀实验(HOS)以及精子染色质扩散(SCD)实验,观察不同浓度薯蓣多糖体外对精子存活率、精子膜完整性以及精子核DNA的影响。结果:6份精液样本孵育24 h及48 h后各组的精子存活率均有降低。维生素C组精子存活率分别为(28.5±3.1)%、(6.5±1.2)%;薯蓣多糖低、中、高剂量组的精子存活率分别为(31.3±3.5)%、(6.5±2.2)%,(37.1±3.5)%、(9.5±2.8)%,(38.3±3.3)%、(9.0±3.2)%,均显著高于生理盐水组[(13.4±4.1)%、(3.1±2.0)%,P0.01],中、高剂量薯蓣多糖组显著高于维生素C组(P0.05及P0.01)。精子DNA碎片率低剂量薯蓣多糖组为(29.4±2.6)%,与维生素C组[(30.5±2.5)%]无明显差异,但显著低于对照组[(41.7±2.2)%]和生理盐水组[(42.1±3.3)%];中、高剂量薯蓣多糖组精子DNA碎片率分别为(28.5±2.3)%和(27.9±1.9)%,显著低于对照组、生理盐水组及维生素C组(P0.05及P0.01)。结论:薯蓣多糖具有体外提高精子存活率和保护精子DNA完整性的作用。     

Induction of oxidative stress by organic hydroperoxides in testis and epididymal sperm of rats in vivo

The present study describes the extent and pattern of oxidative stress induction in testis and epididymal sperm of rats following in vivo exposure to repeated sublethal doses of 2 model pro-oxidants, namely, t-butyl hydroperoxide (tbHP) and cumene hydroperoxide (cHP). Single sublethal (1/40, 1/20, and 1/10 LD(50)) doses of hydroperoxides (HP) administered intraperitoneally to male rats (CFT-Wistar strain) failed to induce any significant increase in malondialdehyde or reactive oxygen species (ROS) levels in testis or epididymal sperm. However, repeated doses for 1 or 2 weeks induced a marked dose-related enhancement of lipid peroxidation (LPO) and ROS levels in both testis and epididymal sperm. Further evidence, such as significant perturbations in both enzymic and nonenzymic antioxidants and enhanced levels of protein carbonyls in testis, suggested induction of oxidative stress. In testis, moderate depletion in reduced glutathione levels and marked diminution in ascorbic acid and alpha-tocopherol content were accompanied by increased activities of various antioxidant enzymes, namely glutathione peroxidase, glutathione-S-transferase, and catalase, in both the HP treatments. Furthermore, significant alterations in the specific activities of testicular enzymes such as LDH-X, G-6-PDH, and SDH indicated altered testicular physiology. Both HP at higher doses induced significant DNA damage (determined by fluorimetric analysis of DNA unwinding assay) in testis and epididymal sperm. Increased total iron levels in testis of HP-treated rats are indicative of the possible involvement of iron-mediated free radical reactions in this model. These findings provide an account of early oxidative damage in testis and epididymal sperm following short-term exposure to HP in vivo, and this model is being further exploited for understanding the consequences of chronic oxidative stress-mediated alterations for the physiology of male reproductive system and its implications for fertility.     

改进型实验性左侧精索静脉曲张的建立及其对大鼠卵泡刺激素及抑制素B的影响

目的:探讨改进型实验性左侧精索静脉曲张(ELV)对大鼠血清卵泡刺激素(FSH)及抑制素B(InhB)的影响。方法:利用改进的方法建立青春期SD大鼠左精索静脉曲张的模型30只;假手术组SD大鼠30只作对照组。术后3个月,取大鼠血清,ELISA法分别检测FSH、InhB的浓度。结果:实验组大鼠血清中FSH浓度[(37.56±9.72)ng/ml]与对照组[(26.69±5.33)ng/ml]相比升高,具有统计学意义(P<0.05);而实验组大鼠血清中InhB的浓度[(349.93±99.48)pg/ml]与对照组[(768.83±146.96)pg/ml]相比下降,具有统计学意义(P<0.05)。结论:改进型ELV致FSH升高及InhB下降,这些变化可能是影响生育能力的机制之一。