黄芪对大鼠睾丸扭转/复位模型保护作用的研究

目的:探讨黄芪注射液对大鼠睾丸扭转/复位模型的保护作用。方法:将30只健康雄性Wistar大鼠分为3组。分别为假手术对照组(A组,n=10);睾丸扭转/复位组(B组,n=10);睾丸扭转/复位+腹腔内注射黄芪注射液组(C组,n=10)。按Turner法建立睾丸扭转模型,喂养至术后7d处死,切取扭转侧睾丸检测凋亡指数(AI)及谷胱甘肽过氧化物酶活力及丙二醛含量。结果:A、B、C三组扭转侧睾丸AI分别为5.82±1.21、36.18±8.40、20.39±3.57,B、C组明显高于对照组(P(0.05),B组明显高于C组(P(0.05)。A、B、C三组扭转侧睾丸谷胱甘肽过氧化物酶活力分别为48.03±2.01、30.93±1.25、38.44±1.06U/mg;丙二醛含量分别为1.43±0.17、3.98±0.36、2.57±0.53nmol/ml,三组之间比较均有显著性差异(P(0.05)。结论:黄芪注射液可明显减少扭转侧睾丸生殖细胞凋亡,保护谷胱甘肽过氧化物酶活力,减轻脂质过氧化程度。     

黄芪注射液对大鼠扭转复位后睾丸组织的保护作用

目的:探讨黄芪注射液对雄性Wistar大鼠扭转复位后睾丸的保护作用。方法:30只大鼠随机分为假手术组(A组)、睾丸扭转复位组(B组)、黄芪注射液治疗组(C组),每组10只,Turner法建立单侧睾丸扭转复位模型,原位缺口末端标记法检测各组睾丸组织中生殖细胞凋亡,化学比色法测定超氧化物歧化酶(SOD)和丙二醛(MDA)含量。结果:黄芪注射液治疗组与睾丸扭转复位组比较,SOD含量明显升高,MDA含量明显降低,生精细胞凋亡指数明显降低。睾丸扭转复位组、黄芪注射液治疗组与假手术对照组比较,SOD含量明显降低,MDA含量明显升高,生精细胞凋亡指数明显升高。结论:黄芪注射液可减少大鼠睾丸扭转复位后睾丸组织的双侧睾丸生殖细胞凋亡,对扭转复位后睾丸生殖细胞有保护作用。其机理可能与提高抗氧化酶活性及减少氧自由基的产生从而减轻大鼠睾丸扭转复位后的缺血再灌注损伤有关。     

单侧隐睾睾丸组织中胰岛素样因子3基因表达

目的 研究单侧隐睾对睾丸组织胰岛素样因子3（Insl3）表达的影响。方法 通过建立隐睾大鼠动物模型获得A组单侧隐睾大鼠12只和B组双侧隐睾大鼠10只，C组空白对照正常大鼠10只。在大鼠8周龄时取睾丸组织用Tarazol法提取总RNA，Northern Blot方法检测各组睾丸组织中Insl3基因表达水平；同时提取各组睾丸组织蛋白，Westernblot检测Insl3蛋白多肽表达。结果 Insl 3 mRNA在单侧隐睾侧睾丸组织中表达量低于C组（P〈0．05），单侧隐睾大鼠对侧睾丸表达量低于C组（P〈0.05）；InSl3基因表达的蛋白多肽在隐睾侧睾丸组织中表达量较低（P〈0．05），对侧睾丸组织蛋白表达较C组低（P〈0．05），但较隐睾侧睾丸和B组明显增多（P〈0．05）。结论 单侧隐睾生精功能的损害可能是影响了隐睾侧睾丸Insl3基因表达，同时还抑制对侧睾丸的表达。     

大鼠单侧睾丸损伤对对侧睾丸组织的影响及其机制的探讨

目的：研究大鼠单侧睾丸损伤后对侧睾丸组织变化及其发生机制。方法：选择SD雄性大鼠40只,随机分为4组：A组为对照组,B组为单侧睾丸挫伤组,c组为单侧睾丸裂伤组,D组为裂伤后腹腔注射环孢素A（cyclosporinA,CsA）组。分别于第2、4周观察大鼠后足垫反应（footpadreaction,FPR）,取对侧睾丸做苏木精一伊红（HE）染色和免疫组化检测CD4分子、CD8分子的表达。结果：注射睾丸抗原48h后,B、C组大鼠右足足垫较A、D组显著增厚（P〈0．01）,C组与B组差异亦有显著统计学意义（P〈0．05）,A组与D组差异无统计学意义（P〉0．05）。B、C组对侧睾丸组织学发生明显改变,A、D组改变不明显。B、C组中CU’T细胞平均光密度（oD）值高于CD。’T细胞（P〈0．05）,两种T细胞OD值均明显高于对照组（P〈0．01）,C组与B组比较,CD4＋T细胞OD值差异有统计学意义（P〈O．05）,CD4＋T细胞OD值差异无统计学意义（P〉0．05）。各组结果在第2、4周比较,差异均无统计学意义。结论：单侧睾丸损伤可引起对侧睾丸组织发生迟发型超敏反应（delayedtypehypersensitivity,DTH）,主要由CD4＋T细胞介导,且与睾丸损伤程度有关。CsA可抑制DTH的发生。     

低温对大鼠睾丸扭转复位后生殖细胞凋亡的影响

目的 探讨低温对大鼠睾丸扭转复位后生殖细胞凋亡的影响。方法 24只健康青春期SD雄性大鼠随机分为三组:A组为睾丸扭转组,B组为睾丸扭转加低温组,C组为对照组。建立单侧睾丸扭转模型。术后第14天采集睾丸。原位缺口末端标记法(TUNEL)检测其生殖细胞凋亡指数(AI),光镜下观察睾丸组织学变化。结果 B组AI明显低于A组(P<0.01),而高于C组(P<0.01)。结论 低温能够提高扭转睾丸耐缺血能力,减少睾丸扭转复位后生殖细胞凋亡。     

大鼠单侧睾丸扭转复位后对对侧睾丸的影响

目的研究大鼠单侧睾丸扭转复位后对对侧睾丸的影响。方法16只成年健康SD雄性大鼠随机分为实验组(n=8)和对照组(n=8)；建立单侧睾丸扭转复位模型。术后30d取扭转对侧睾丸,采用原位缺口末端标记法(TUNEL)检测生殖细胞凋亡,光镜下计数精子数。结果与对照组相比,实验组对侧的睾丸重量和日产精子量都有显著性差异(P＜0.05),实验组生殖细胞凋亡显著增多(P＜0.01)。结论大鼠单侧睾丸扭转后,对侧睾丸生殖细胞凋亡增多可能是导致不育的原因之一。     

低温联合地塞米松对大鼠睾丸扭转复位后ICAM1表达的影响及生精功能的保护作用

目的:探讨低温联合地塞米松对大鼠睾丸扭转复位后的生精功能的保护作用,以及对细胞间粘附分子1(ICAM1)表达的影响。方法:将100只青春期的雄性SD大鼠(体重140~160 g)随机分为4组,每组25只。4组大鼠分别扭转左侧睾丸720°2 h,建立单侧睾丸扭转动物模型,各组做如下处理,A组:常温+生理盐水;B组:低温+生理盐水;C组:常温+地塞米松;D组:低温+地塞米松;术后48 h采集睾丸,通过HE染色光镜观察睾丸组织病理学改变、TUNEL法检测睾丸生精细胞的凋亡、Western印迹检测ICAM1的表达。结果:HE染色光镜下见4组大鼠扭转侧睾丸组织均有不同程度损伤,其中A组睾丸损伤最为明显,其余3组扭转侧睾丸得到不同程度保护;睾丸组织ICAM1 Western印迹检测结果:A组扭转侧(左侧)睾丸组织ICAM1蛋白表达量(0.68±0.03)高于B组(0.49±0.06)、C组(0.46±0.09)、D组(0.17±0.08),差异具有显著性(P0.05、P0.05、P0.01);凋亡细胞染色:细胞核呈深棕黄色或棕褐色,A组扭转侧睾丸可见大量生精细胞凋亡,凋亡指数AI[(33.13±3.21)%]明显高于B组[(17.12±5.23)%](P0.05)、C组[(14.13±2.03)%](P0.05)及D组[(9.05±1.03)%](P0.01)。结论:低温联合地塞米松能显著增强睾丸组织抗损伤能力,较好地保护扭转复位后睾丸的生精功能及降低ICAM1的表达。     

低温联合地塞米松对大鼠睾丸扭转复位后eNOS表达及生精细胞凋亡的影响

目的:探讨低温联合地塞米松对睾丸扭转复位后的保护作用,以及对eNOS表达及生精细胞凋亡的影响。方法:将80只青春期SD大鼠随机分为4组,每组20只。4组大鼠分别扭转左侧睾丸720°2 h,建立单侧睾丸扭转模型,随后各组做如下处理,A组:常温+生理盐水、B组:低温+生理盐水、C组:低温+地塞米松、D组:常温+地塞米松;术后48 h采集睾丸,通过HE染色光镜观察睾丸组织病理学改变、免疫组化法检测eNOS表达、TUNEL法检测睾丸生精细胞凋亡。结果:HE染色光镜下见4组大鼠扭转侧睾丸组织均有不同程度损伤,其中A组睾丸损伤最明显,其余3组扭转侧睾丸得到不同程度保护;睾丸组织eNOS免疫组化检测结果:A组扭转侧(左侧)睾丸组织阳性细胞数及阳性细胞着色强度明显强于B、C、D 3组,差异具有显著性(P<0.05、P<0.01、P<0.01);凋亡细胞染色:细胞核呈深棕黄色或棕褐色,A组扭转侧(左侧)睾丸可见大量生精细胞凋亡,凋亡指数AI(31.12±4.68)明显高于B组(16.58±6.22)(P<0.05)及C(8.60±1.15)、D组(13.52±3.06)(P<0.01)。结论:睾丸扭转复位后的缺血再灌注损伤可导致生精细胞凋亡增加、睾丸生殖能力下降;应用低温联合地塞米松能显著增强睾丸组织的抗损伤能力,较好地保护了扭转复位后睾丸的生精功能。     

大鼠睾丸扭转生殖细胞凋亡与抗氧化酶和Bcl-2基因表达的关系

目的　探讨睾丸扭转复位后生殖细胞凋亡及其发生机制。方法　建立单侧睾丸扭转复位大鼠模型(72 0° ,2h)。术后 72小时取手术侧睾丸 ,采用TUNEL法检测生殖细胞凋亡 ,免疫组化SP法检测Bcl 2基因表达 ,并测定睾丸组织中超氧化物歧化酶 (SOD)和过氧化氢酶 (CAT)活性。结果　实验组手术侧睾丸生殖细胞凋亡增多 ,Bcl 2基因低表达 ,SOD和CAT活性下降 ,与对照组相比 ,两组差别具有极显著性意义 (P <0 .0 1)。结论　睾丸扭转复位后生殖细胞凋亡增多与Bcl 2基因低表达和抗氧化酶活性下降有关。     

【原创论文】同侧和对侧大鼠睾丸缺血再灌注损伤的生化效应和褪黑激素对此的保护作用

睾丸扭转（TT）,是一个多发于青春期男性的泌尿急症,如果不及时治疗,可致不孕不育。睾丸扭转所致缺血再灌注（I／R）损伤在睾丸损伤的病理生理过程中起一定作用。我们研究了褪黑激素在单侧睾丸扭转大鼠中同侧和对侧睾丸的氧化损伤效应：将21只青春期雄性Wistar大鼠分成三组,每组七只,处理如下：第1组（假手术组）：行左睾丸和双边睾丸假切除术;第2组（I/R组）：通过以下方式诱发缺血再灌注损伤（顺时针720°旋转左侧睾丸2小时,2小时后复位）：第3组（I／R＋MEL组）：大鼠经诱导缺血再灌注损伤和一次性褪黑激素注射（50mgkg-1,i．p）。处理后离体分离各组大鼠双侧睾丸,用于检测睾丸组织中抗氧化过氧化氢酶、超氧化物歧化酶和谷胱甘肽过氧化物酶的活性,丙二醛、蛋白质羰基和一氧化氮的组织水平。较对照组,褪黑激素注射组同侧睾丸的脂质过氧化水平,相关酶活性降低,具有显著性（P〈0．05）,而在对侧的睾丸中相关酶活性变化无统计学意义（P〉0．05）。在对侧睾丸中,丙二醛水平改变具有明显统计学意义（P=0．009）。应用褪黑素能减轻老鼠同侧睾丸扭转所致缺血再灌注损伤的不利影响,而对于对侧睾丸,睾丸扭转影响不大。     

Protective role of erythropoietin during testicular torsion of the rats

Testicular torsion is an important clinical urgency. Similar mechanisms occurred after detorsion of the affected testis as in the ischemia reperfusion (I/R) damage. This study was designed to investigate the effects of erythropoietin (EPO) treatment after unilateral testicular torsion. Fifty male Sprague-Dawley rats were divided into five groups. Group 1 underwent a sham operation of the right testis under general anesthesia. Group 2 was same as sham, and EPO (3,000 IU/kg) infused i.p., group 3 underwent a similar operation but the right testis was rotated 720° clockwise for 1 h, maintained by fixing the testis to the scrotum, and saline infused during the procedure. Group 4 underwent similar torsion but EPO was infused half an hour before the detorsion procedure, and in group 5, EPO was infused after detorsion procedure. Four hours after detorsion, ipsilateral and contralateral testes were taken out for evaluation. Treatment with EPO improved testicular structures in the ipsilateral testis but improvement was less in the contralateral testis histologically, but EPO treatment decreased germ cell apoptosis in both testes following testicular IR. TNF-α, IL-1β, IL-6 and nitrite levels decreased after EPO treatment especially in the ipsilateral testis. We conclude that testicular I/R causes an increase in germ cell apoptosis both in the ipsilateral and contralateral testes. Eryhropoietin has antiapoptotic and anti-inflammatory effects following testicular torsion.     

New animal model to evaluate testicular blood flow during testicular torsion.

BACKGROUND/PURPOSE: Unilateral testicular torsion is known to cause infertility because of damage to the contralateral testis. Testicular damage has been attributed to many different mechanisms, one of which is altered contralateral blood flow. In our experiment, in an effort to identify the reason for contralateral testicular injury, the authors developed an accurate method of measuring blood flow in both testes before, during, and after unilateral torsion. METHODS: Four- to 6-week-old piglets weighing 4 to 6 kg were studied. The animals were anesthetized, intubated, ventilated, and catheterized for vascular access. Piglets were assigned randomly to a sham group or a group undergoing 360 degrees or 720 degrees torsion of the left testis (n = 5 per group) for 8 hours, after which it was untwisted. Data were collected at baseline (T = 0), 8 hours of torsion (T = 8), and 1 hour after detorsion (T = 9). Mean arterial blood pressure and heart rate were monitored continuously. Testicular blood flow was determined using radiolabeled microspheres. Blood flow data were evaluated by analysis of variance. RESULTS: In the 360 degrees torsion group, blood flow changes were insignificant during torsion and after detorsion. In the 720 degrees torsion group, blood flow to the twisted testis was reduced significantly, whereas the contralateral testis was unaffected. One hour after detorsion, blood flow to both testes was increased significantly. CONCLUSIONS: The authors describe a new animal model to evaluate testicular blood flow during and after testicular torsion. Increased blood flow after detorsion may be the cause of testicular damage in patients with unilateral testicular torsion.     

Reperfusion injury after detorsion of unilateral testicular torsion

Summary Reperfusion injury has been well documented in organs other than testis. An experimental study was conducted to investigate reperfusion injury in testes via the biochemical changes after unilateral testicular torsion and detorsion. As unilateral testicular torsion and varicocele have been shown to affect contralateral testicular blood flow, reperfusion injury was studied in both testes. Given that testicular blood flow does not return after 720° testicular torsion lasting more than 3 h, the present study was conducted after 1 and 2 h of 720° torsion. Adult male albino rats were divided into seven groups each containing ten rats. One group served to determine the basal values of biochemical parameters, two groups were subjected to 1 and 2 h of unilateral testicular torsion respectively, two groups were subjected to detorsion following 1 and 2 h of torison respectively, and two groups underwent sham operations as a control. Levels of lactic acid, hypoxanthine and lipid peroxidation products were determined in testicular tissues. Values of these three parameters obtained from the sham operation control groups did not differ significantly from basal values (P>0.05). All three parameters were increased significantly in both ipsilateral and contralateral testes after unilateral testicular torsion when compared with basal values (P<0.01 and P<0.05, respectively). Detorsion caused significant changes in lipid peroxidation products levels in ipsilateral but not in contralateral testes when compared with values obtained after torsion (P<0.01 and P>0.05, respectively). It is concluded that ipsilateral testicular torsion causes a decrease in perfusion not only in the ipsilateral but also in the contralateral testis. Additionally, detorsion following up to 2 h of 720° torsion causes reperfusion injury in ipsilateral but not in contralateral testis.     

Effect of unilateral testicular torsion on blood flow and histology of contralateral testes.

BACKGROUND/PURPOSE: Infertility occurs in 25% of patients after unilateral testicular torsion; hence, the authors examined hemodynamic and histological changes in both testes after acute testicular torsion in neonatal piglets. METHODS: The animals were anesthetized, intubated, ventilated, catheterized, and assigned randomly to a sham group or one of three experimental groups undergoing 720 degrees torsion of the left testis for 8 hours after which it was untwisted in group I and removed in group II. In group III, both testes were removed. Data were collected at baseline (T = 0), 4 hours (T = 4), and 8 hours of torsion (T = 8) and at the ninth hour of the experiment (T = 9). Testicular blood flow was determined by using radiolabeled microspheres. The testes also were examined blindly with routine and electron microscopy. RESULTS: In group I, testicular blood flow decreased in the affected testis during torsion and increased significantly after detorsion, whereas blood flow to the contralateral testis increased significantly after detorsion. Sham-operated animals showed no histological abnormality in either testis. In all torsion groups, the affected testis showed extensive changes caused by hemorrhagic necrosis. The contralateral testis only showed changes in group I. CONCLUSION: Unilateral testicular torsion resulted in ipsilateral damage caused by a decrease and subsequent increase in blood flow while in the contralateral testis; damage was the result of a significant increase in blood flow after detorsion.     

Blood perfusion of the contralateral testis evaluated with contrast-enhanced ultrasound in rabbits with unilateral testicular torsion

The changes of blood perfusion of contralateral testis after unilateral testicular torsion remain controversial. In this study, 28 New Zealand white male rabbits were randomly divided into five groups. Group A (n = 8), the control group, underwent a sham operation on the unilateral testis without inducing testicular torsion. In groups B, C, and D (n = 5 each), unilateral testicular torsion was induced, and, after 3, 6 or 24 h, respectively, detorsion was performed. In group E (n = 5), permanent unilateral testicular torsion was applied. Contrast-enhanced ultrasound was used to observe the blood perfusion of the contralateral testis at the following stages: pre-torsion (preopration), immediately post-torsion (postopration), pre-detorsion, immediately post-detorsion, and late-stage post-detorsion (6–12 h post-detorsion in groups B–D) or at a similar time point (15–21 h post-torsion in group E). Time-intensity curves were generated, and the following parameters were derived and analyzed: arrival time, time to peak intensity, peak intensity, and half-time of the descending peak intensity. The analysis revealed that blood perfusion of the contralateral testis increased immediately after testicular torsion on the opposite side (P < 0.05), which increased with prolonged testicular torsion of the other testis. This research demonstrated that contrast-enhanced ultrasound was valuable in evaluating blood perfusion of the contralateral testis after unilateral testicular torsion.     

Experimental testicular torsion and its effects on the contralateral testicle

Objective Following experimental unilateral torsion of the testis the histologic effects of unilateral testicular torsion on the contralateral testis were investigated. Materials and methods Utilizing detorsion or orchiectomy at 4 hours and 8 hours after torsion, the effects of early and late treatment modalities on the contralateral testicle were observed. Results Morphometry of the contralateral testis revealed some alterations including focal sclerosis, decrease in mean seminiferous tubular diameter and a marked increase of the Leydig cells in some subgroups. Conclusion In spite of some changes, definite evidence for contralateral damage due to ipsilateral torsion contributing to male infertility was hardly observed.     

The role of nitric oxide in testicular ischemia-reperfusion injury

PURPOSE: This study was designed to determine the role of nitric oxide (NO) in the ischemia-reperfusion (I/R) injury process in testes. METHODS: Fifty prepubertal male rats were divided into 5 groups each containing 10 rats. After 4-hour torsion and 4-hour detorsion, bilateral orchiectomies were performed for measurement of tissue malondialdehyde (MDA) level and histopathologic examination. The results were compared statistically. The groups were labeled as group 1, basal values of biochemical parameters in testes; group 2 (control group), torsion plus detorsion; group 3, torsion plus N-monomethyl-L-arginine (L-NMMA) plus detorsion; group 4, torsion plus L-arginine plus detorsion; group 5, sham operation. RESULTS: The highest MDA values were determined in the L-arginin group in ipsilateral testes. Group 3 and group 4 were statistically different from control group. Histological examination showed that specimens from group 4 had a significantly (P < .05) greater histological injury than group 3, and contralateral testes showed normal testicular architecture in all groups. CONCLUSIONS: These results suggest that NO plays an important role in damaging the testis with I/R. Although inhibition of NO synthesis with L-NMMA significantly improves I/R injury in testes, enhancing NO production by providing excess of L-arginine increases such damage. In the early periods of detorsion, there is no damage to contralateral testes after unilateral testicular torsion.