生脉注射液对不同周龄大鼠睾丸扭转复位后睾丸损伤影响的研究

目的:探讨生脉注射液对不同周龄大鼠睾丸扭转复位后睾丸损伤影响的差异。方法:3、6、12周龄健康雄性SD大鼠各16只,随机分成睾丸扭转复位+注射生脉注射液组(实验组)和睾丸扭转复位+注射生理盐水组(对照组),每组8只,建立睾丸扭转动物模型(左侧阴囊切开,绕精索顺时针扭转睾丸720°2h),并于手术后24h处死,测定各组大鼠睾丸组织内总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性与丙二醛(MDA)含量。结果:与各自对照组比较,3、6周实验组大鼠左侧睾丸组织中T-AOC、SOD活性和MDA含量均无显著性改变(P>0.05);12周实验组大鼠左侧睾丸组织中SOD、T-AOC明显升高,而MDA含量显著降低,差异均具有显著性(P<0.05)。结论:生脉注射液对睾丸扭转复位后的缺血再灌注急性损伤有保护作用,但对不同周龄大鼠的再灌注损伤保护作用不同,存在年龄相关性差异,对较大周龄大鼠的急性保护作用较为明显。     

大鼠单侧精索扭转后保留同侧坏死睾丸对对侧睾丸和附睾的组织学影响

目的:将大鼠单侧精索持续扭转96 h,研究保留扭转侧坏死睾丸对对侧睾丸和附睾的影响,以说明延迟性的睾丸切除是否对对侧睾丸功能有保护作用。方法:将33只青春前期(21～42日龄)正常雄性SD大鼠,随机分为假手术对照组(n=11)、扭转保留组(n=12)和扭转切除组(n=10)。假手术对照组只对左侧睾丸行睾丸肉膜囊固定术,后两组扭转实验组用睾丸肉膜囊固定术固定维持扭转720°的睾丸、附睾,在扭转96 h后将扭转保留组扭转侧睾丸、附睾行复位及固定,而同时扭转切除组则将扭转侧睾丸、附睾切除。术后3个月抽取血液标本,ELISA测定大鼠血清睾酮、抗精子抗体浓度。同时取睾丸、附睾标本,石蜡包埋切片后作组织学观察,并利用体视学方法定量研究睾丸、附睾结构的体积以及生精小管直径。结果:3组大鼠血清睾酮值无统计学差异;仅扭转保留组造模后1例大鼠血清抗精子抗体阳性。光镜下定性观察发现精索扭转组睾丸间质细胞核较假手术对照组间质细胞核增大,假手术对照组、扭转保留组和扭转切除组睾丸结构有明显形态改变的数量分别为1、3、0只。扭转保留组对侧睾丸相对假手术对照组睾丸体积增加19%,相应附睾体积增加11%,扭转切除组对侧睾丸体积增加21%,附睾体积增加7%,且睾丸代偿性肥大具有统计学意义(P<0.05)。假手术对照组、扭转保留组和扭转切除组对侧睾丸生精小管体积分别为(1.15±0.07)、(1.30±0.04)、(1.35±0.05)cm3,3组间无显著性差异。3组大鼠对侧睾丸内间质体积分别为(0.25±0.02)、(0.36±0.02)、(0.34±0.03)cm3,精索扭转组和假手术对照组相比显著增加(P<0.05)。3组大鼠对侧睾丸内生精小管直径分别为(226.00±7.00)、(223.00±6.00)、(221.00±3.00)μm,无显著性差异(P>0.05)。结论:单侧精索长期扭转后对侧睾丸、附睾主要改变是单侧去势后的对侧代偿性肥大表现,是否切除对对侧睾丸和附睾组织学上无明显影响。     

大鼠一侧睾丸扭转对侧睾丸改变的实验研究

目的 :研究一侧睾丸扭转 (UTT)后对侧睾丸组织学及生精细胞凋亡的改变 ,以明确UTT后对侧睾丸是否存在损伤。　方法 :SD雄性大鼠 6 0只 ,随机分为实验组 (n =4 8)及对照组 (n =12 )。实验组采用Turner方法建立左侧睾丸扭转模型 ,于扭转后 6h处死 4只 ,其余 4 4只再分为扭转睾丸复位及切除组 ,分别于术后 1d、1周、4周处死7～ 8只 ,取睾丸组织进行组织学及生精细胞凋亡的检测。　结果 :UTT复位后对侧睾丸组织学发生明显改变 ,生精细胞凋亡指数明显高于对照组 (P <0 .0 5 )。扭转睾丸切除后对侧睾丸变化不明显。　结论 :UTT可引起对侧睾丸损伤 ,其机制可能与再灌注有关 ,扭转睾丸切除可防止或减轻对侧睾丸的损伤     

高压氧对睾丸扭转/复位后生精细胞凋亡的作用

目的 探讨单侧睾丸扭转/复位后睾丸细胞凋亡状况和高压氧(HBO)的干预作用。方法 32只青春期 Wistar大鼠随机等分为 4组,1组为对照组,2～4组制成单侧睾丸扭转/复位模型(扭转 720°、2 h后复位),3组、4组分别于睾丸复位前和复位后立即予以 60 min HBO治疗。术后48 h获取双侧睾丸,原位缺口末端标记法(TUNEL)观察细胞凋亡,常规染色观察病理学改变。结果 与1组比较,2组扭转侧睾丸生精细胞凋亡增加(P<0.01)、病理损害明显;对侧睾丸生精细胞凋亡增加(P<0.01)。3～4组尤其是4组扭转侧睾丸生精细胞凋亡较2组明显减少(P<0.01)、病理损害减轻;对侧睾丸细胞凋亡较1组无明显变化(P>0.05)。结论 单侧睾丸扭转复位后,双侧睾丸生精细胞凋亡增加,复位前和复位后早期HBO治疗能明显减少细胞凋亡的发生。     

地塞米松对大鼠睾丸扭转复位后保护作用的研究

目的探讨地塞米松对大鼠睾丸扭转复位后的保护作用.方法24只成年健康SD雄性大鼠随机分为3组A组为睾丸扭转复位加生理盐水组,B组为睾丸扭转复位加地塞米松组,C组为对照组.建立单侧睾丸扭转复位模型.术后24h取手术侧睾丸,化学比色法测定睾丸组织的SOD、CAT、MPO活性和MDA含量,光镜观察病理学变化.结果与A组相比,B组的MDA含量和MPO活性降低(P＜0.01),SOD和CAT的活性增高(P＜0.01),睾丸被膜下白细胞浸润减少,无间质水肿.结论地塞米松可减轻大鼠睾丸扭转复位后的再灌注损伤,对睾丸有保护作用.     

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。     

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。     

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。     

大鼠睾丸扭转复位后附睾唾液酸含量变化及意义

目的 :探讨大鼠睾丸扭转 2h和 4h复位后 2 4h附睾唾液酸含量的变化和意义。　方法 :用 2 4只雄性SD大鼠建立左侧睾丸扭转复位模型 ,分为对照组、扭转 2h组和 4h组 ,每组 8只。 5 甲基苯二酚法检测扭转侧附睾唾液酸的含量。　结果 :睾丸扭转 2h复位后 2 4h扭转侧附睾唾液酸含量 [(2 3.385± 9.2 2 0 )mg/mgprot]改变不明显 ;睾丸扭转 4h复位后 2 4h附睾唾液酸含量 [(13.72 5± 7.80 1)mg/mgprot]下降明显 (P <0 .0 5 )。　结论 :睾丸扭转 2h复位后 2 4h附睾分泌唾液酸功能不受影响 ,扭转 4h复位后 2 4h附睾分泌唾液酸功能下降 ;附睾耐受缺血再灌注损伤的时间可能较长。     

一氧化氮在一侧睾丸扭转对侧睾丸损伤中的作用

目的 研究总抗氧化能力(T-AOC)和一氧化氮(NO)在一侧睾丸扭转对侧睾丸损伤中的作用。方法 SD雄性大白鼠建立左侧睾丸扭转模型，于扭转后6h再分为扭转睾丸复位及切除组，分别于术后1h、1d、1周、2周和4周处死4—5只，取出睾丸用于一氧化氮合酶(NOS)活性、NO、T-AOC及细胞凋亡的检查。结果 UTT复位后对侧睾丸组织NOS活性、NO含量明显升高，T—AOC显著降低。结论 NO过量产生及T-AOC的下降是UTT对侧睾丸损伤的重要原因。     

【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。     

The protective effects of nitric oxide on the contralateral testis in prepubertal rats with unilateral testicular torsion

OBJECTIVE: To investigate histological changes in the contralateral testis of rats with unilateral testicular torsion and the protective effects of nitric oxide (NO) on possible damage. MATERIAL AND METHODS: Twenty-eight prepubertal male Sprague-Dawley rats were divided into four equal groups. Group 1 underwent a sham operation of the right testis under general anaesthesia. Group 2 underwent a similar operation but the right testis was rotated 720 degrees clockwise for 6 h, maintained by fixing the testis to the scrotum, and saline infused during the procedure. Group 3 underwent similar torsion but L-arginine methyl ester (a precursor of NO) was infused during the procedure. In Group 4, NG-nitro-L-arginine-methyl ester, a NO synthase inhibitor, was infused separately during the administration of L-arginine methyl ester and torsion. All the left (untwisted) testes were removed from rats 21 days after surgery and evaluated histologically, assessing seminiferous tubule diameter, loss of sperm and spermatids, loss of germ cell layers, disarray of germ cell layers, rupture of tubules, Leydig cell proliferation and reaction in the ruptured tubules, and oedema. RESULTS: There was a significant difference in the indicators of histological damage between groups 2 and 4 and groups 1 and 3, except for the Leydig cell reaction in the ruptured tubules and oedema. The damage was significantly less in group 3 than in groups 2 and 4. CONCLUSION: These results suggest that long-term histopathological changes in the contralateral testes are important after unilateral testicular torsion and that NO has a protective effect on the contralateral testis.     

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。     

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.     

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.     

The effect of testicular torsion on the contralateral testis and the value of various types of treatment

In an attempt to investigate the effect of testicular torsion and various forms of treatment on the contralateral testis, an experimental study on rats was undertaken. The first group comprised control animals. In the second group the left testes were twisted 720 degrees and the right testes were removed 4 weeks later for histopathological examination. In the third group the rats were subjected to a left detorsion procedure 24 h after torsion, while in the fourth group cortisone treatment was added to the above procedure. The fifth group consisted of rats which had undergone left orchiectomy 24 h after torsion and the sixth group had cortisone treatment plus orchiectomy after torsion. Cortisone treatment was started 24 h after testicular torsion and continued for 4 weeks. Histopathological examination of the contralateral testes which were removed 4 weeks later showed that either orchiectomy plus cortisone or detorsion plus cortisone was more successful than other forms of treatment.     

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.     

单侧睾丸扭转对生殖细胞凋亡及黄芪保护作用的实验研究

目的观察大鼠单侧睾丸扭转／复位后患侧和对侧睾丸生精细胞凋亡情况,探讨单侧睾丸扭转／复位后生殖能力下降的机制以及黄芪注射液对其再灌注损伤的保护作用。方法将40只健康雄性Wistar大鼠分为4组,分别为假手术对照组（A组）,睾丸扭转／复位组（B组）,睾丸扭转／复位＋单次腹腔内注射黄芪注射液组（C组）及扭转／复位十连续腹腔内注射黄芪注射液组（D组）,每组10只。按Turner法建立睾丸扭转／复位模型,所有大鼠均在同等条件下喂养至术后7d处死,切取双侧睾丸后检测凋亡指数。结果扭转侧睾丸生殖细胞凋亡指数（AI）A组（5．82±1．21）与B组（36．18±8．40）、C组（20．39±3．57）、D组（11．61±5．12）相比差异有显著性（P〈0．05）,B组明显高于C组及D组（P〈0．05）,C组与D组相比差异有显著性（P〈0．05）;B组对侧睾丸（12．95±3．06）与C组（9．45±1．71）、D组（7．56±1．06）两组对侧睾丸AI相比差异有显著性（P〈0．05）,C、D两组对侧睾丸AI差异有显著性（P〈0．05）。结论单侧睾丸扭转可致患侧和对侧睾丸生精细胞凋亡明显增加,黄芪注射液可明显减少双侧睾丸生殖细胞凋亡,连续应用黄芪注射液优于单次应用。     

Capsaicin effectively prevents apoptosis in the contralateral testis after ipsilateral testicular torsion

OBJECTIVE: To evaluate the effect of afferent nerve blockage by administration of capsaicin on apoptotic changes in the contralateral testis in rats undergoing ipsilateral testicular torsion. MATERIALS AND METHODS: Twenty male albino rats were randomly divided into four groups. In groups 1 and 2, rats underwent a sham operation and testicular torsion, respectively, after the intraperitoneal administration of 0.9% NaCl. Similarly, in groups 3 and 4 the rats underwent a sham operation and testicular torsion, respectively, after an intraperitoneal capsaicin injection. The testes were untwisted 24 h later and the contralateral testes harvested. Apoptosis was assessed in paraffin-embedded sections stained for nuclear DNA fragmentation. Fifteen cells were counted in each seminiferous tubule and the apoptotic cells recorded. A score was calculated for each group and the results compared using the Kruskal-Wallis analysis of variance and Mann Whitney U-tests, with P<0.05 considered to be significant. RESULTS: The mean apoptotic score of group 2 was significantly higher than that of the other groups. There was no difference between the apoptotic scores of groups 1 and 3, 1 and 4, and 3 and 4. CONCLUSION: Capsaicin effectively prevented apoptosis in the contralateral testes of rats that had undergone testicular torsion.