Testicular tissue nitric oxide and thiobarbituric acid reactive substance levels: evaluation with respect to the pathogenesis of varicocele

The aim of the present study is to evaluate tissue nitric oxide (NO) and thiobarbituric acid reactive substance (TBARS) levels in testicular tissue, and to determine their relationship with seminal parameters in order to explain possible effects on varicocele pathophysiology.Ten adult male Wistar rats at 8 weeks old underwent partial left renal vein ligation. A sham operation was performed on control rats in a second group of another ten rats. All animals were killed 4 weeks after surgery. The testes were removed and histological changes were observed by light microscopy with haematoxylin and eosin stain on half of each testis. The rest of testis was used for the evaluation of testicular tissue NO and TBARS levels. Epididymal aspirated seminal plasma was used for semen analysis and morphological analysis was carried out according to Krugers criteria. Statistical analysis was performed by using Mann-Whitney U-tests and Spearman rank correlations between the two groups for NO and TBARS levels and for seminal parameters. Testicular tissue NO and TBARS levels (mean±SEM) were 62.8±10.1 mol/g protein and 4.7±0.3 nmol/g protein in group 1. These parameters were 16.9±2.2 mol/g protein and 3.1±0.2 nmol/g protein in the group 2 controls. There were significant differences between these parameters (P_NO=0.000, P_TBARS=0.001). Although a positive and significant correlation between testicular tissue NO and TBARS levels was found (r_s=0.739, P=0.014), there was only a strong negative correlation between NO levels and sperm motility in group 1 (r_s=–0.815, P=0.004). We found that this effect of NO on sperm motility was independent from TBARS levels after regression analysis (r²=-0.687, =0.825, P=0.034). Although there were statistically significant differences in seminal parameters between the two groups, there was no difference between them in the histopathological examination. We found that sperm motility was significantly related to testicular tissue NO levels only. Thus, we suggest that NO is an important mediator in the pathogenesis of varicocele. TBARS and other substances have been effective via NO pathways.     

亚临床型精索静脉曲张患者精浆NO浓度与精子活力关系

目的 探讨亚临床型精索静脉曲张(SVC)精浆一氧化氮(NO)浓度与精子活力的关系.方法 选择亚临床型精索静脉曲张患者35例为实验组,同时选择正常健康男性35例作对照组.采用计算机辅助精液分析技术测定各组精子活力,采用硝酸还原酶法测定精浆中NO浓度.结果 ①实验组精浆中NO浓度为52.19±17.12μmol/L,对照组精浆中NO浓度为38.06±6.43 μmol/L,实验组明显高于对照组,两者比较差异具有统计学意义(P＜0.05).②实验组精浆中NO水平与精子活力水平呈负相关性,具有统计学意义(P＜0.05).结论 亚临床型精索静脉曲张患者精浆中NO浓度增高,并在一定范围内与精子活力呈负相关.     

Immunocytochemical distribution of nitric oxide synthase in the human seminal vesicle: a light and electron microscopical study

Although nitric oxide (NO) has been proven to be one of the most important non-adrenergic, non-cholinergic mediators in the control of human reproductive tract organs, to date information on the significance of NO-mediated signal transduction in the control of human seminal vesicle (SV) function is still sparse. Recent investigations have underlined the significance of NO in the maintenance of sperm capacitation and viscosity of the seminal plasma as well as in the control of mammalian seminal vesicle smooth muscle tone. In order to further investigate the functional impact of NO on the regulation of normal SV function, we examined the distribution of NADPH-diaphorase (NADPH-d), endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS) in the cellular anatomy of human SV by means of light and electron microscopical immunocytochemistry (LM, EM) in combination with the tyramide signal amplification technique. Human SV were obtained from 15 patients who had undergone surgery for pelvic malignancies (carcinoma of the prostate or urinary bladder). SV specimens were fixed, sectioned and examined by LM and EM for the presence of NAPDH-d, eNOS and nNOS using specific antibodies and advanced staining procedures. LM revealed a dense NADPH-d reaction in glandular epithelial structures, whereas no substantial labeling was detected in the fibromuscular stroma. EM showed that the NADPH-d reaction product was abundantly detectable attached to membranes of the endoplasmic reticulum, mitochondria and the nuclei of glandular epithelial cells. nNOS staining was found in nerve fibers branching within the SV tissue. eNOS staining was present in small vessels but was only observed to a minor degree in glandular and subglandular structures and the smooth muscle stroma. Our results support the hypothesis that human SV is a site of NO production. The distribution of NADPH-d may give rise to the speculation that NO is mainly involved in the regulation of SV secretory activity. The sparse correlation between NADPH-d-, eNOS- and nNOS-staining might hint at the existence of a previously unidentified NOS isoform in human SV.     

门静脉高压症时细菌移位与内毒素血症和一氧化氮之间关系的实验研究

目的 探讨门静脉高压症时细菌移位与内毒素血症和一氧化氮（NO）之间的关系。方法 采用门静脉部分缩窄（PVS）制成门静脉高压模型的30只大鼠均分为3组：模型组（B）、左旋精氨酸组（C）、单－甲基精氨酸组（D）。另10只大鼠接受假手术作为对照组（A）。术后2周取肠系膜淋巴结（MLN）、脾和血标本进行细菌培养;测定门静脉压力、内毒素和NO2^-水平;并用镧做示踪剂观察肠粘膜屏障通透性变化和粘膜 形态的改变。结果 PVS各组MLN细菌检出率高于A组,血浆内毒素水平均上升。PVS各组NO2^-和门静脉压力均较A组增高,在D组二者均较B组下降。细菌移位率、内毒素和NO2^-水平之间呈显著正相关。镧沉积到PVS各组的粘膜上皮细胞和其上下固有层细胞间,粘膜细胞的微绒毛有形态改变。结论 门静脉高压症可发生细菌移位的内毒素血症,可能与肠粘屏障通透性增强和破坏有关,提示细菌移位引起的内毒素血症并增加NO2^－水平。     

肝脏缺血再灌注损伤与一氧化氮和内皮素平衡关系的研究

目的　研究一氧化氮 (NO)与内皮素 (ET)平衡关系 (NO/ET)的变化与肝脏缺血再灌注(I/R)损伤的关系。方法　采用鼠肝I/R模型 ,并应用不同工具药物观察血NO/ET比值的变化以及肝损伤的情况。结果　I/R急性期血NO/ET比值降低 (1.5 8± 0 .2 0至 0 .2 9± 0 .0 5 ,P <0 .0 1) ,肝损伤加重。NO供体L 精氨酸 (L Arg)与ET受体拮抗剂TAK 0 44在一定程度上可减轻肝I/R损伤 ,而NO合酶抑制剂L NAME进一步加重了损伤。结论　肝I/R损伤与NO/ET平衡关系破坏有关 ,调节其平衡关系可影响I/R损伤的程度。     

一氧化氮与大鼠缺血性急性肾衰竭关系的实验研究

目的 研究一氧化氮（ＮＯ）在缺血性急性肾衰竭病理过程中的作用。方法 通过夹闭大鼠双侧肾蒂４５分钟后再松夹复制出急性肾衰（ＡＲＦ）模型,各组在松夹后分别静滴生理盐水,Ｌ－精氨酸,Ｄ－精氨酸,Ｎ－硝基－Ｌ－精氨酸（ＮＬＡ）。结果 与盐水对照组相比,Ｌ－精氨酸组菊糖清除率和再灌注早期的尿流率增高（Ｐ〈０．０５）,尿钠排泄分数下降（Ｐ〈０．０５）,肾病理损害也较轻（Ｐ〈０．０５）,ＮＬＡ虽升高血压（Ｐ〈０     

一氧化氮和一氧化氮合成酶在急性肺损伤的作用

急性肺损伤(acute lung injury,ALI)是一种发病率和死亡率都很高的常见临床疾病. 目前,对ALI病理生理学基础和临床研究方面的了解越来越多,但并没有提出新的治疗策略能够明显改善ALI的治疗.在ALI的动物模型和患者中,一氧化氮合成酶(nitric oxide synthases,NOS)表达及活性增强和一氧化氮(NO)的增多在ALI的病理生理过程中有重要作用;但临床抑制NO生成以及选择性抑制NOS并没有对ALI的治疗有明显效果.目前提出了不同细胞源性NO的概念,这种NO的细胞源性差异可能对ALI的治疗有潜在的意义.现综述NO和NOS在ALI中的作用.     

脑出血患者一氧化氮、一氧化氮合酶的变化及临床意义

目的 探讨一氧化氮(NO)、一氧化氮合酶(NOS)与脑出血的关系。方法 测定76例脑出血患者及66例健康人血浆NO、NOS含量，分析上述指标在出血后3个时期的变化以及与患者颅内血肿量、是否合并蛛网膜下腔出血和临床预后的关系。结果 与对照组相比，脑出血1～3d组NO、NOS含量明显高于4～7d组、14d组和对照组，NO、NOS变化与颅内血肿量和／或合并蛛网膜下腔出血有一定关系。结论 NO、NOS参与了脑出血病理生理过程，其变化对病情的预后有一定的意义。     

动脉硬化闭塞症与内皮素、一氧化氮的关系

目的 探讨内皮素（ＥＴ）、一氧化氮（ＮＯ）在动脉硬化闭塞症（ＡＳＯ）形成和发展听 作用及ＥＴ、一氧化氮合酶（ＮＯＳ）基因表达同ＡＳＯ的关系。方法 收集４０例ＡＳＯ病人、１２０处病变血管和３０例正常血管,根据病理学检查分为ＡＳＯ早期、中期、晚期和正常血管４组。利用放射免疫测定法和Ｇｒｉｓｓｅ方法检测ＡＳＯ病脸的血浆ＥＴ和ＮＯ水平,利用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术检测４组血管ＥＴｍＲＮＡＴ     

脾亢患者血小板计数与脾动脉血流量及血清一氧化氮水平间关系

目的 探讨脾亢患者血小板计数与脾动脉血流量及血清一氧化氮(NO)水平间关系.方法 肝硬化伴脾亢患者36例(脾亢组),健康体检者10例(对照组).血清NO浓度测定用硝酸还原酶法,脾动脉血流量由彩色多普勒超声仪测量.结果 脾亢组和对照组血小板计数分别为(60.75±27.77)×10~9/L、(212.40±71.76)×10~9/L(P<0.01);脾动脉血流量分别为(482.50±263.61)、(181.20±61.85)ml/min(P<0.01);血清NO水平分别为(97.646±20.990)、(50.774±19.400)μmol/L(P<0.01).脾亢组血小板计数与脾动脉血流量显著负相关(r=-0.422,P<0.05),血清NO水平与脾动脉血流量呈显著正相关(r=0.746,P<0.01).结论 血清NO水平升高导致脾动脉血流量增加可能在脾亢发生发展中起着重要作用.     

一氧化氮合酶抑制剂对创伤性休克的干预作用

目的：评价选择性诱导型一氧化氮合酶（iNO）抑制剂氨基胍（AG）和非选择性一氧化氮合酶抑制剂L-N位硝基精氨甲酯（L-NAME）对创伤性休克的治疗效果。方法：40只Wistar大鼠制作创伤性休克动物模型。双侧股骨干砸伤后并经股动脉放血至平均动脉压（MAP）35～45mmHg（4．67～6.00kPa），维持30min，然后回输失血和等量的林格式液。随机分为休克组（10只）,AG组（根据复苏时静脉注射AG含量为10，50mm／kg。则分为AGⅠ，AGⅡ各10只），L-NAME组（10只，复苏时静脉注射LNAME10mg/kg），观察休克前后血浆NO浓度的动态变化及24h大鼠存活率。并留取肺、肝、肾、小肠组织，观察病理改变。结果：大鼠创伤性休克后，血浆NO水平明显高于休克前；AG各组动物复苏后血浆NO的水平明显降低，各脏器的病理损害亦显著减轻，存活率明显提高，AGⅡ组效果最好；L-NAME组动物复苏后血浆NO的水平也明显降低，各脏器的病理损害无明显变化，存活率无明显提高。结论：NO在创伤性休克的病理发展过程中起着重要作用，应用AG有助于创伤性休克的改善；L-NAME能降低NO的水平，但对休克的预后无明显改善。     

Increased vasoconstrictor reactivity and decreased endothelial function in high grade varicocele; functional and morphological study

The pathophysiology of human varicocele is not fully understood. We investigated vasoconstrictor reactivity, endothelial function and morphological changes in different grades of varicocele to clarify the pathophysiology. Contractile responses to phenylephrine, norepinephrine, serotonin and histamine were determined in isolated human varicose spermatic veins using the organ bath technique. Endothelial function was tested with acetylcholine-induced relaxation after phenylephrine-induced precontraction in the absence and presence of nitric oxide synthase inhibitor, L-NAME, and cyclooxygenase inhibitor, indomethacin. The cyclic guanosine monophosphate (cGMP) level was measured in the spermatic vein and peripheral plasma. Morphological changes were evaluated with light microscopy. Phenylephrine, norepinephrine, serotonin and histamine induced concentration-dependent contractions. The maximum contractions for all of these agents except norepinephrine were significantly higher in grade III than grade I and II (P<0.05). The sensitivity to phenylephrine was significantly higher in grades II and III than in grade I (P<0.05). In the presence of L-NAME and indomethacin, the difference from respective control phenylephrine-induced contractions was higher in grade I and II than grade III. Acetylcholine did not induce stable relaxation but the level of cGMP, which is responsible for the vasorelaxant effect of NO, in veins was lower in grades II and III than grade I (P<0.05). Vessel wall thickness increased in grade II and dilatation developed in grade III when compared to grade I (P<0.05). Our findings suggest that endothelium produces less vasorelaxant which results in the more enhanced effects of vasoconstrictor substances in grade III, indicating that endothelial dysfunction develops at high grades of varicocele.     

比较局麻与全麻对病人血清NOS及NO浓度的影响

目的　比较局麻与全身麻醉对手术病人血清一氧化氮合酶 (NOS)和一氧化氮 (NO)的影响。方法　根据不同麻醉方法将 2 0例颌面外科手术病人分为局麻组 (A组 )和静吸复合全麻组 (B组 ) ,每组 10例 ,在麻醉诱导前、手术切皮和手术开始后 1h抽取静脉血 3ml,分别测定血清NOS和NO浓度。结果　A组血清NOS及NO含量在切皮和术中 1h与术前比较显著增高 (P <0 0 5 ) ,B组血清NOS及NO含量在切皮和术中 1h与术前比较无显著增高 (P >0 0 5 ) ;在切皮和术中 1hB组NOS及NO含量比A组显著降低 (P <0 0 5 )。结论　静吸复合麻醉能明显抑制NOS的活性 ,降低NO的生成     

颅内动脉瘤术后脑血管痉挛与血浆内皮素-1、一氧化氮含量的关系

目的　探讨内皮素 (ET) 1、一氧化氮 (NO)与颅内动脉瘤术后脑血管痉挛发生的关系。方法　颅内动脉瘤 3 0例 ,经颅多普勒 (TCD)检测脑血流速度 ,并抽取外周静脉血测定血浆ET 1和NO含量 ,进行手术前后对比研究。另选取 10例健康成人进行对比研究。结果　 (1)脑血管痉挛组术后 1～ 3d、5～ 7d血浆ET 1的平均含量明显高于同期未发脑血管痉挛组 (P <0 .0 1) ,与术前和正常组相比差异亦有显著性 (P <0 .0 1)。 (2 )脑血管痉挛组术后 1～ 3d血浆NO的平均含量明显低于术前 (P <0 .0 5 )和术后 5～ 7d(P <0 .0 1) ,术后 5～ 7d脑血管痉挛组的血浆NO平均含量明显高于未发生脑血管痉挛组和正常组 (P <0 .0 1)。结论　 (1)颅内动脉瘤术后脑血管痉挛的发生与血浆ET 1、NO平均含量呈明显相关性。 (2 )ET 1/NO在维持脑血管张力上起着重要作用 ,当两者动态平衡破坏时 ,可能导致脑血管痉挛发生。 (3 )对于术后血浆ET 1含量明显升高和 /或NO含量明显降低的患者应及时行经颅多普勒检查 ,有助于尽早发现脑血管痉挛 ,使脑缺血、脑梗死得到及时的预防和治疗     

Significant correlation between cerebrospinal fluid nitric oxide concentrations and neurologic prognosis in incomplete cervical cord injury

In animal models of spinal cord injury (SCI), inducible NO (nitric oxide) synthase is expressed in the spinal cord immediately after sustaining SCI. Excessive NO production has cytotoxic effects and induces neuronal apoptosis, causing neural degeneration and neurodysfunction in the spinal cord. Little is known, however, about the relationship between NO_x (NO metabolites: nitrite and nitrate) levels in the cerebrospinal fluid (CSF) and neurologic severity or recovery in clinical cases. The objective of the present study was to examine the correlation between CSF NO_x levels and neurologic severity or recovery in SCI. Twenty-five patients with incomplete cervical cord injury (CCI) were examined. Eight cases were treated conservatively (non-operated group). Seventeen cases underwent surgical intervention (operated group). NO_x levels in the CSF were measured using the Griess method. The severity of the neurologic impairment was assessed using Frankel’s classification and the American Spinal Injury Association motor score (ASIA MS). The degree of neurologic recovery was assessed using Frankel’s classification and the ASIA motor recovery percentage (MRP). There was no significant difference in the NO_x levels between the CCI group (NO_x levels: 5.9 ± 0.7 μM) and the 36 control subjects (1 volunteer and 35 patients without neurologic disorders, NO_x levels: 4.9 ± 0.3 μM). There was no significant difference in NO_x levels and MRP between the non-operated group and the operated group. The NO_x levels in total SCI group were significantly correlated with the ASIA MS and MRP. There was a significant correlation between CSF NO_x levels and neurologic severity or recovery in incomplete CCI.     

去势成年雌性大鼠骨质疏松模型血清NO和N0S的水平及其意义

目的 测定大鼠骨质疏松模型血清中NO和NOS的水平,探讨骨质疏松症与NO和NOS浓度的关系。方法 6月龄成年雌性SD大鼠随机分成实验组和对照组,术前及术后均测量腰椎骨密度,术后分离血清,用改良镀铜镉还原法测定NO含量,用化学比色法测定NOS含量。结果 术前,两组大鼠骨密度差异无显著性,术后,实验组大鼠骨密度明显低于对照组骨密度,差异有显著性(P<0.05)。血清NO和NOS水平实验组明显低于对照组,差异有显著性(P<0.05),有统计学意义。结论NO和NOS在骨质疏松的病理生理过程中起重要作用。     

一氧化氮在获得性肾囊肿癌变过程中的作用

目的　探讨一氧化氮 (NO)过多产生在获得性肾囊肿 (ACDK)发生肾癌过程中的作用。　方法　用高压液相自动分析方法测定 1 6例 1 8份ACDK和 1 2份单纯性肾囊肿囊内液体中NO的代谢产物亚硝酸盐 /硝酸盐 (NO-2 /NO-3 )的含量 ,同时测定 5份ACDK患者血浆NO-2 /NO-3 的含量。采用免疫组织化学方法 (LSAB法 )检测 1 7份ACDK和 2 0例非ACDK肾组织一氧化氮合酶 (iNOS)的表达。　结果　ACDK囊内液NO-2 /NO-3 的含量 (1 5 1 .6± 6 4.2 μmol/L)明显高于单纯性肾囊肿组(5 0 .1± 33.6 μmol/L) ,差异有极显著性 (P <0 .0 0 1 )。其中在相同的 5例ACDK患者中囊内液NO-2 /NO-3 的浓度明显的高于血浆中的浓度。在免疫组织化学染色 (LSAB法 )检测肾组织iNOS的表达中 ,1 7份ACDK中 1 4例 (82 % )为阳性。 1 3例正常肾组织和 7例非ACDK慢性肾衰的肾组织均无i NOS的表达。　结论　NO在ACDK囊内液中过多产生有可能是其囊上皮发生肾癌的原因之一。     

生育男性和异常精子症病人精浆一氧化氮水平的差异

目的 :通过研究精子正常和异常男性精浆中一氧化氮 (NO)含量及动态的差异 ,以了解NO与男性生育力的关系。　方法 :采用硝酸还原酶法 ,经Greiss试剂显色后 ,用分光光度法测定 174例生育男性和 2 17例异常精子症男性精浆中NO的含量。　结果 :① 174例精子生育男性精浆中均检测出NO ,平均含量为 (2 7.78± 5 .81) μmol/L,并随年龄增加而增高 ,40岁以后增高显著。 2 0～ 2 9岁组 [(2 6 .2 5± 5 .5 2 ) μmol/L]与 30～ 39岁组 [(2 8.11± 5 .87)μmol/L]相比 ,差异无显著性 (P >0 .0 5 ) ,但 40～ 49岁组 [(30 .17± 6 .14) μmol/L]与 2 0～ 2 9岁组相比 ,增高有统计学意义 (P <0 .0 5 )。② 9类精子异常症男性的精浆中NO含量均显著高于精子正常组。增高幅度以精子单项异常为最小 ,以精子的 3项异常为最高 ,2项精子异常的精浆中NO水平介于两者之间。　结论 :男性精浆中适度的NO可能具有调节精子发生的作用。     

冠状动脉旁路术围术期肺动脉血浆一氧化氮合成酶活性与一氧化氮浓度的变化

目的 研究冠状动脉旁路术围术期肺动脉血浆一氧化氮合成酶（NOS）活性和一氧化氮（NO）浓度的变化。方法 选择30例冠状动脉旁路手术（CABG）病人,在围术期抽取肺动脉血测定NOS和NO值。结果 肺动脉血浆中NOS和NO水平虽有明显变化（P＜0．05）,但NO浓度与NOS活性的增减并不一致：当NOS活性降低时,NO浓度增高;反之当NOS活性显著增高时,NO浓度降低。结论 在行CABG围术期中,肺动脉血浆中内源性NO分泌无法满足应激时自身保护对其的需要。     

内皮素、NO与自体静脉移植再狭窄的关系

目的　探讨内皮素、 Ｎ Ｏ 同自体静脉移植后狭窄的关系。方法　对２６ 例自体静脉移植后狭窄并进行２ 次手术者以及正常对照组２６ 例,利用放射免疫测定、 Ｇｒｉｓｓ 方法检测血浆内皮素和 Ｎ Ｏ 水平,利用反转录聚合酶链反应检测狭窄血管组织内皮素和一氧化氮合酶（ Ｎ Ｏ Ｓ） 基因表达水平。结果　所有患者均存在高内皮素、低 Ｎ Ｏ 血症,存在狭窄血管组织内皮素基因高表达、 Ｎ Ｏ Ｓ 基因低表达状态。结论　内皮素、 Ｎ Ｏ 同自体静脉移植再狭窄密切相关。