茶多酚对大鼠睾丸扭转/复位模型保护作用的研究

目的:探讨茶多酚对大鼠睾丸扭转/复位模型的保护作用。方法:将24只健康雄性Wistar大鼠随机分为3组,每组8只。第Ⅰ组为假手术组(切开左侧阴囊游离睾丸,但不予扭转),第Ⅱ、Ⅲ组扭转左侧睾丸720°6h,分别于扭转复位前30min腹腔注射生理盐水和茶多酚,术后连续3d分别以低剂量维持。3组大鼠喂养至术后第5天处死,切取左侧扭转睾丸检测睾丸组织中超氧化物歧化酶(SOD)和丙二醛(MDA)含量;以原位缺口末端标记法(TUNEL)检测生精细胞凋亡指数(AI)。结果:Ⅰ、Ⅱ、Ⅲ3组左侧扭转睾丸组织SOD活力分别为(285.00±22.51)、(242.00±17.62)、(261.00±10.01)nU/mg;MDA含量分别为(1.81±0.20)、(4.34±0.34)、(2.94±0.38)nmol/mg;3组之间比较均有显著性差异。Ⅰ、Ⅱ、Ⅲ3组左侧扭转睾丸生精细胞凋亡指数(AI)分别为6.64±1.82、55.23±6.46、31.84±5.56,第Ⅲ组与第Ⅱ组相比,其生殖细胞凋亡明显减少(P<0.05)。结论:茶多酚对因睾丸扭转导致的缺血再灌注损伤具有保护作用。     

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。     

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。     

葡萄籽原花青素对小鼠睾丸扭转复位后生精功能的保护作用

目的:探讨葡萄籽原花青素(GSP)对小鼠睾丸扭转复位后生精功能的保护作用。方法:24只健康雄性昆明小白鼠(8周龄,25～27 g)随机分为3组:对照组、扭转组、治疗组,每组8只。扭转组及治疗组建立单侧睾丸扭转复位动物模型,治疗组于扭转复位前30 min腹腔注射GSP(50 mg/kg),术后采用腹腔注射方式连续给药3 d,每天1次,每次50 mg/kg。扭转组方法同治疗组,治疗同体积生理盐水。术后第4天取扭转侧睾丸,检测组织病理学参数和生精细胞凋亡指数(AI),并检测睾丸组织超氧化物歧化酶(SOD)和丙二醛(MDA)含量。对照组行假手术。结果:治疗组与扭转组相比,Johnsen评分上升[(7.38±0.92)分vs(5.00±1.85)分,P<0.05],生精小管直径略增大[(178.75±1.58)μm vs(176.50±1.60)μm,P>0.05],生精细胞层数增加[(5.75±0.71)层vs(3.75±1.03)层,P<0.05],生精细胞凋亡指数AI明显降低[(16.25±1.67)%vs(40.50±1.60)%,P<0.05)],SOD活性明显上升[(52.67±3.57)U/mg prot vs(29.04±4.46)U/mg prot,P<0.05],MDA含量明显下降[(2.91±0.04)nmol/mg prot vs(4.63±0.05)nmol/mg prot,P<0.05]。结论:GSP对小鼠睾丸扭转复位后生精功能损伤有明显的保护作用,其作用机制可能与其能清除氧自由基、抑制脂质过氧化、提高机体抗氧化能力有关。     

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。     

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。     

己酮可可碱对大鼠睾丸扭转复位后生精功能的保护

目的:探讨己酮可可碱(PTX)对大鼠睾丸扭转复位后生精功能的保护作用。方法:雄性SD大鼠24只,随机分成3组,每组8只,建立睾丸扭转动物模型。第Ⅰ组为假手术组(扭转720°后立即复位),第Ⅱ、Ⅲ组扭转720°2 h,于复位前15 m in分别静脉注射生理盐水和PTX,术后24 h留取手术侧睾丸。应用流式细胞术(FCM)检测各组生精细胞凋亡和各级生精细胞计数,采用硫代巴比妥酸法测定丙二醛(MDA)含量,化学比色法测定组织内总抗氧化能力(T-AOC)。结果:第Ⅲ组应用PTX后与第Ⅱ组相比,生精细胞凋亡明显减少(399.50±33.31vs1221.75±132.48,P<0.01),单倍体和四倍体细胞群计数显著增多(5554.13±441.28vs4102.35±206.98;1906.00±200.72vs1711.63±144.55,P均<0.01;),T-AOC明显回升(32.52±2.86vs22.76±3.73,P<0.01),MDA含量下降(1.78±0.20vs3.98±0.36,P<0.01),其差异均有显著性(P<0.01)。结论:己酮可可碱对睾丸扭转复位后的生精功能具有明显的保护作用。     

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。     

灯盏花素对青春前期大鼠睾丸扭转复位双侧睾丸的保护作用

目的:探讨灯盏花素青春前期大鼠睾丸扭转复位双侧睾丸的远期保护作用。方法:对建立左侧睾丸扭转复位动物模型,将32只4周龄健康SD雄性大鼠随机分为4组:假手术组、生理盐水组(腹腔内注射生理盐水2 mg/kg)、灯盏花素单次给药组(睾丸复位前30 min,腹腔缓慢注入灯盏花素2 mg/kg,给药体积为0.5 ml/100 g体重)和灯盏花素连续给药组(在单次给药组的基础上,于术后每天注射同等剂量灯盏花素注射液1次,连续7 d),每组8只。术后6周处死大鼠,取双侧睾丸和附睾,检测睾丸组织总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性与丙二醛(MDA)含量,测定精子浓度、存活率和活动率,行睾丸组织病理学观察。结果:与同侧生理盐水组比较,两给药组双侧SOD、T-AOC、NOS活性、精子浓度、精子存活率和精子活动率均升高,MDA含量下降,其中各检测指标除单次给药组健侧T-AOC活性、精子浓度、精子活动率、双侧NOS活性及连续给药组健侧NOS活性外差异均显著(P0.05);与同侧单次给药组比较,连续给药组双侧SOD、T-AOC、NOS活性、精子浓度、精子存活率和精子活动率均升高,MDA含量下降,其中各项指标除健侧NOS活性外差异均显著(P0.05)。生理盐水组可见双侧生精小管退变,间质水肿,且患侧受损明显大于健侧;两给药组使双侧睾丸扭转复位后损伤的组织学改变明显改善。结论:灯盏花素可通过有效清除氧自由基,减轻脂质过氧化损伤,对青春前期大鼠睾丸扭转复位双侧睾丸有明显的保护作用,且连续给药明显优于单次给药。     

环孢素对大鼠睾丸缺血-再灌注损伤后生精细胞凋亡的影响

目的探讨环孢素对大鼠单侧睾丸缺血-再灌注损伤后生精细胞凋亡的影响,研究其保护性作用。方法雄性SD大鼠24只,随机分成假手术组、扭转组和环孢素组,每组8只,建立720°2h单侧睾丸扭转动物模型。扭转组和环孢素组于复位前15min分别腹腔注射生理盐水和环孢素,术后24h留取手术侧睾丸。应用流式细胞术检测各组患侧睾丸组织生精细胞凋亡;Quantitative Real-time PCR技术对Fas、FasL和Bax mRNA进行定量分析;Western-blot技术检测细胞色素C含量。结果与假手术组相比,扭转组患侧睾丸组织早期凋亡生精细胞百分比明显增多,Fas、FasL和Bax mRNA表达上调,同时胞质中细胞色素C含量显著升高,其差异均有显著性(P均<0.05)。环孢素干预能显著减轻上述变化,患侧睾丸组织正常细胞群百分比升高,Fas、FasL和Bax mRNA表达下调,同时胞质中细胞色素C含量显著降低,其差异均有显著性(P均<0.05)。结论环孢素可能参与调控生精细胞凋亡的分子途径,对睾丸扭转术后的生精功能具有明显的保护性效果。     

单侧睾丸扭转后生精细胞凋亡的分子途径

目的:研究大鼠单侧睾丸扭转复位后生精细胞凋亡的分子机制。方法:雄性SD大鼠16只,随机分为对照组和扭转组,每组8只。建立睾丸扭转动物模型(720°2h),术后24h留取手术侧睾丸。应用流式细胞术检测生精细胞凋亡和各级生精细胞计数,应用RT-PCR技术对Fas/FasL mRNA和Bax mRNA进行半定量分析,Western印迹技术检测细胞色素C含量。结果:两组间生精细胞凋亡及各类生精细胞计数均有显著性差异(P<0.01)。扭转组FCM直方图呈现高大凋亡峰,单倍体和四倍体细胞群计数下降,Fas/FasL mRNA和Bax mRNA表达上调,同时细胞质中细胞色素C含量亦明显升高,与对照组相比其差异均有显著性(P均<0.01)。结论:睾丸扭转复位后生精细胞凋亡存在着外源性和内源性两条基本途径。凋亡相关分子Fas/FasL表达上调和Bax介导的细胞色素C释放可能是睾丸扭转后生精细胞凋亡的重要环节。     

Role of endogenous cannabinoids in ischemia/reperfusion injury following testicular torsion in rats

Objectives:   The role of endogenous cannabinoids in ischemia/reperfusion induced germ cell apoptosis in rats was investigated.
Methods:   Baseline group was for basal normal values. The Sham operated group served as a control group. The torsion/detorsion (T/D) group underwent torsion (1 h) and detorsion; AN1, AN2, and AN3 groups received anandamide (10 mg/kg) 30 min before torsion, 30 min after torsion, and just after detorsion, respectively. In the AM251 group, AM251 (0.5 mg/kg) was injected 45 min before torsion and in the AN/AM group, AM251 and anandamide were injected 45 and 30 min before torsion, respectively. Lipid peroxidation, antioxidant enzymes, and germ cell apoptosis was determined.
Results:   Malondialdehyde (MDA) levels in the T/D group were significantly higher than the control group. Moreover, MDA values in the AN1, AN2, and AN3 groups were significantly lower than T/D. There were significant decreases in catalase and superoxide dismutase activities in the T/D group versus the control group. These values in the AN1, AN2, and AN3 groups were significantly higher than T/D. It was also shown that MDA levels in the AN/AM group were significantly higher than the AN1 group. In the AN/AM group, catalase and superoxide dismutase activities were significantly lower versus the AN1 group. The mean germ cell apoptosis scores in all animals with testicular T/D were significantly higher than the control group. There was no difference between the apoptotic indices in the AN1, AN2, AN3, and T/D groups. Apoptosis scores in AM251 and AN/AM were significantly higher compared with the T/D and AN1 groups.
Conclusions:   Although anandamide increased antioxidant markers, it failed to reduce germ cell apoptosis. AM251 worsened the antioxidant defense system, which is reflected as higher germ cell apoptosis.     

Effect of insulin-like growth factor-1 on apoptosis of rat testicular germ cells induced by testicular torsion

OBJECTIVE: To investigate the possible protective role of insulin-like growth factor-1 (IGF-1, reported to have a protective effect in experimental models of hypoxic ischaemia), and the involvement of apoptotic cell death in a model of torsion/detorsion of the rat testis. MATERIALS AND METHODS: Adult male Wistar rats were divided into five groups of five rats each. Group 1 underwent a sham operation as a control; in group 2 the testis was twisted and in group 3 then untwisted; in group 4 IGF-1 was injected subcutaneously just before bilateral torsion, and then the right testis removed after 4 h and the left after 24 h; in group 5, IGF-1 was injected immediately after bilateral detorsion and then the testes removed as in group 4. Both testicles were examined histologically, with apoptosis detected using the in situ DNA fragmentation (TUNEL) system, with combined enzymology and immunohistochemistry techniques. RESULTS: In groups 2 (torsion) and 3 (detorsion), light microscopy of the testis showed some degenerative changes in the germ cells. Compared to group 1, apoptosis was more significant in group 3 than in the other groups. Group 4 (torsion/IGF-1) had a similar number of apoptotic germ cells as in group 2 (torsion) after 24 h, but fewer than the same group after 4 h. In group 5 (detorsion/IGF-1), apoptosis was reduced by IGF-1 significantly more than in group 3 (P < 0.05). Apoptosis was significantly less in spermatids in group 5 than in group 3 (P < 0.05). CONCLUSIONS: IGF-1 seems to lower the levels of germ cell apoptosis, which may be important for protecting the testes from torsion/detorsion injury. Further clinical studies are needed to evaluate this protective effect in testicular torsion/detorsion.     

The protective effect of Cold-inducible RNA-binding protein (CIRP) on testicular torsion/detorsion: An experimental study in mice

Purpose

To evaluate the expression of Cold-inducible RNA-binding protein (CIRP) in torsion/detorsion of the testes in different phases and demonstrate the protective effect of CIRP on testicular injury after torsion/detorsion (T/D) in an experimental mouse model.

Methods

Twenty-four male BALB/c mice were divided randomly into 8 groups: normal control group (N), sham-operated group (S), torsion 2 h group (T2h), torsion/detorsion 12 h group (T/D12h), and T/D24h, T/D48h, T/D72h, and T/D96h groups. The testes were examined for the expression levels of CIRP. Another 32 male BALB/c mice were divided randomly in to 4 groups: normal control group (N), T/D group, T/D + pcDNA3.1 group, and T/D + pcDNA3.1-CIRP group. The plasmids were transfected into testes with in vivo-jetPEI. After 3 days, morphological changes, mean seminiferous tubule diameter (MSTD), and the number of the germ cell layers were observed. Superoxide dismutase (SOD) activity, the levels of malondialdehyde (MDA), and Bcl-2/Bax ratios were studied in the different groups.

Results

Compared with the N and S groups, the expression of CIRP in the T2h group was down-regulated. In T/D groups, the levels of CIRP were reduced in a time dependent manner. Compared to T/D and T/D + pcDNA3.1 group, the MSTD, number of the germ cell layers, SOD activity, and Bcl-2/Bax ratio increased in T/D + pcDNA3.1-CIRP group, while the level of MDA decreased.

Conclusions

The results of our study have shown that down-regulated CIRP is involved in testicular injury after testicular torsion/detorsion. Up-regulation of the expression of CIRP may reduce the damage caused by torsion/detorsion, possibly by preventing germ cell oxidative stress and apoptosis.     

Protective effects of immunophilin ligands on testicular torsion/detorsion damage in rats

Objectives  The purpose was to investigate the role of immunophilin ligands in ischemia/reperfusion (I/R)-induced germ cell apoptosis in the rat. Materials and methods  Sprague–Dawley rats were divided into five groups with ten animals in each. In animals undergoing torsion/detorsion, right testes were rotated 720o for 1 h. A baseline group was for basal normal values. The sham-operated group served as a control group. The TD group underwent torsion/detorsion surgery alone; the cyclosporine-A group (TD-CsA) received intravenous cyclosporine injection (5 mg/kg) at the time of detorsion, and the FK-506 group (TD-FK) received intravenous FK-506 (3.5 mg/kg) at the time of detorsion. For measurement of lipid peroxidation and antioxidant enzyme activities, the right testes of five animals in each group were excised after 4-h reperfusion. Germ cell apoptosis indices were determined 24 h following detorsion in the right testes of the remaining five animals in each group. Results  Malondialdehyde (MDA) levels in the TD group were significantly higher compared to control and baseline groups. Moreover, testicular MDA values in TD-CsA and TD-FK groups were significantly lower than in TD. There were also significant decreases in catalase and superxide dismutase activities in the TD group compared to control and baseline groups. These values in TD-CsA and TD-FK groups were significantly higher than in TD. The mean germ cell apoptosis scores were significantly higher in TD animals compared to control and baseline groups; however, CsA and FK-506 treatment significantly reduced the apoptosis compared with the TD group. Conclusion  We have shown that administration of immunophilin ligands in testicular torsion decreases ischemia/reperfusion (I/R) cellular damage. The results of biochemical studies suggest that reduction of oxidative stress along with attenuated neutrophil accumulation by immunophilin ligands may have a major role in their cytoprotective effects.     

睾丸扭转后生精细胞凋亡与iBOS基因表达

本文研究了睾丸扭转复位后生精细胞凋亡与iBOS基因表达的关系。采用大鼠建立左侧睾丸扭转复位模型（720,2h)。用TUNEL法和免疫组化SP法分别检测扭转复位后第五天生精细胞凋亡和iBOS基因表达。研究发现凋亡主要见于染色质降解的生精细胞（初级精母细胞和圆形精子细胞）。间质细胞和支持细胞未见凋亡发生。iBOS表达见于各级生精细胞,在染色质降解的生精细胞（即凋亡细胞）强表达。本文研究表明睾丸扭转复位后生精细胞凋亡增加与iBOS基因表达密切相关。睾丸局部NO生成异常可能是生精细胞凋亡增加的原因之一。