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。     

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。     

维拉帕米及低温对大鼠睾丸扭转复位后生精功能的保护作用

目的:探讨药物维拉帕米及低温对大鼠睾丸扭转复位后生精功能的影响。方法:60只健康青春期SD大鼠随机均分成5组,A组为睾丸扭转组,25℃室温,扭转时间2h;B组为睾丸扭转加维拉帕米组;C组为睾丸扭转加低温(9～13℃)组;D组为睾丸扭转加维拉帕米、低温组;E组为对照组。按Turner法制作睾丸扭转模型,取手术侧睾丸,光镜下观察组织学变化,行生精小管Makler评分,流式细胞术检测凋亡细胞百分率。结果:HE染色:除E组光镜下观察正常外,其余各组生精细胞排列不整齐,层次减少,生精细胞数量减少,可见凋亡小体及坏死区,部分区域可见炎症细胞浸润,其中扭转组最明显。生殖细胞凋亡率:A、B、C、D、E组分别为(32.11±2.20)%,(20.18±1.50)%,(20.02±1.90)%,(13.75±1.40)%,(8.56±0.90)%,与A组比较,B、C、D组凋亡率降低,差异有显著性(P<0.01)。Makler评分结果:A、B、C、D、E组分别为(14.47±1.35)、(15.45±0.75)、(15.48±0.75)、(16.22±0.72)、(19.60±0.56)分,与A组比较,B、C、D组Makler评分升高,差异有显著性(P<0.01)。结论:睾丸扭转复位后生精细胞凋亡增加可导致睾丸生殖能力下降;应用钙通道阻滞剂维拉帕米及局部低温均能增强睾丸组织的抗损伤能力;两者联合应用能更好地保护扭转复位睾丸的生殖功能。     

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。     

DPPH法测定葡萄籽原花青素清除自由基的能力

通过DPPH法测定葡萄籽原花青素抗氧化、清除自由基能力,实验表明:随着葡萄籽原花青素纯化程度的提高,其抗氧化、清除自由基的能力也相应地提高。葡萄籽原花青素纯化物的抗氧化、清除自由基能力大于VC和芦丁,葡萄籽粗提物的抗氧化、清除自由基能力略低于VC,但高于芦丁。GSP、GSPP1、GSPP2、GSPP3、GSPP3-SP、VC、芦丁清除DPPH自由基的半抑制量分别为118,93,88,86,63,110,170 g/kg,其自由基清除能力AE分别为0.84×10-2、1.08×10-2、1.14×10-2、1.16×10-2、1.58×10-2、0.59×10-2、0.59×10-2。     

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。     

睾丸扭转后生精细胞凋亡与iNOS基因表达

本文研究了睾丸扭转复位后生精细胞凋亡与iNOS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iNOS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)。问质细胞和支持细胞未见凋亡发生。iNOS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。本研究表明睾丸扭转复位后生精细胞凋亡增加与iNOS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。     

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。     

葡萄籽提取物原花青素对紫外线照射朗格汉斯细胞的影响

目的:评估外用葡萄籽提取物原花青素对皮肤光损伤后朗格汉斯细胞的影响.方法:实验设计正常皮肤组(G1组)、单纯2MED照射组(G2组),基质+2MED照射组(G3组),含葡萄籽提取物原花青素样品外用+2MED照射组(G4)共四组,连续3天同样处理24小时后对皮肤取材,进行CDla免疫组化染色.结果:G2组朗格汉斯细胞明显减少,G4组朗格汉斯细胞数比正常皮肤略有增多,并且树突明显增多变长.结论:葡萄籽提取物原花青素能够保护皮肤免受紫外线照射所致的免疫抑制,其机制可能与防止紫外线照射所致的朗格汉斯细胞减少有关.     

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。     

The protective effect of darbepoetin alfa on experimental testicular torsion and detorsion injury

AIM: Testicular torsion is a serious urological emergency, usually involving newborns, children, and adolescents which can lead to subfertility and infertility. Prevention of testicular damage caused by torsion is still a clinical and experimental problem. So far many chemicals and drugs have been investigated for decreasing ischemia/reperfusion (I/R) injury in experimental animals. The possible protective effect of darbepoetin alfa, a novel erythropoietic protein, on testicular tissue after I/R injury was examined in this study. METHODS: Thirty rats were divided into three groups: sham operation, torsion/detorsion, and torsion/detorsion plus darbepoetin alfa groups. After torsion (2 hours) and detorsion (4 hours), bilateral orchiectomy was performed. Malondialdehyde, nitric oxide and glutathione levels were determined in testicular tissue. RESULTS: Administration of darbepoetin alfa caused a decrease of malondialdehyde and nitric oxide levels and an increase in glutathione levels compared with the torsion/detorsion group. In addition, histological injury scores were significantly decreased in the treatment group more than the torsion/detorsion group. CONCLUSION: The results suggest that darbepoetin alfa may be a potential protective agent for preventing testicular injury caused by testis torsion.     

Effects of montelukast and zileuton on testicular torsion/detorsion injury in rats

The aim of this study was to evaluate and compare the effects of 5‐lipoxygenase enzyme (5‐LO) inhibitor zileuton and cysteinyl leukotriene receptor (CysLT1R) antagonist montelukast in testicular torsion/detorsion (T/D) injury model in rats. Rats were anaesthetised with 75 mg kg^?1 ketamine hydrochloride and 8 mg kg^?1 xylazine intraperitoneal before the operation. Torsion was created by rotating the right testis 720° clockwise and maintained by fixing the testis. The rats were treated with CysLT1R antagonist montelukast (10 mg kg^?1; i.p.), 5‐LO inhibitor zileuton (3 mg kg^?1; i.p.), and vehicle, at 30 min prior detorsion. After 1 h of torsion, the testis was counter‐rotated to the natural position and replaced into the scrotum. Malondialdehyde (MDA) level was measured in testicular tissue after 3 h of reperfusion. Histological examination was performed after 24 h of reperfusion. T/D caused a significant increase in MDA level and histopathological injury in testes. Montelukast and zileuton treatments prevented the T/D‐induced augmentation in MDA levels. Only zileuton treatment significantly reduced the T/D‐induced histopathological injury. In this study, we demonstrated for the first time that zileuton had protective effects on testicular T/D injury. We have also found that zileuton is more effective than montelukast on histopathological injury.     

Protective effects of sumatriptan on ischaemia/reperfusion injury following torsion/detorsion in ipsilateral and contralateral testes of rat

This study was planned to evaluate the effects of sumatriptan, 5‐HT1B/1D receptors agonist, on ischaemia/reperfusion injury in bilateral testes after unilateral testicular torsion/detorsion in rats. Male Wistar rats (n = 42) were allocated into a sham‐operated group, a control group and treatment groups which were injected sumatriptan (0.1, 0.3 and 1 mg/kg), GR‐127935 (0.01 mg/kg)—5‐HT1B/1D receptors antagonist—and sumatriptan (0.1 mg/kg) + GR‐127935 (0.01 mg/kg). Torsion was induced for 1 hr by rotating right testis 720⁰ in the clockwise direction, and after 7 days of detorsion, bilateral orchiectomy was conducted. While the level of TNF‐α rose in testicular tissue after inducing torsion/detorsion, sumatriptan injection notably lowered TNF‐α level in ipsilateral (torted) and contralateral (nontorted) testes (p < 0.001). Moreover, after inducing testicular torsion/detorsion, SOD activity was decreased, whereas administration of sumatriptan significantly increased SOD activity in bilateral testes (p < 0.001). After induction of torsion/detorsion, macroscopic and histological analyses also showed severe damages which were improved by sumatriptan injection. Interestingly, co‐administration of sumatriptan with GR‐127935 reversed the beneficial impacts of sumatriptan on macroscopic appearance, microscopic pattern and biochemical markers. It is concluded that sumatriptan presumably via stimulation of 5‐HT_1B/1D receptors decreased inflammation, oxidative stress and deteriorations induced by ischaemia/reperfusion injury following testicular torsion/detorsion.     

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。     

“生精散”对大鼠睾丸扭转复位后生精功能的影响及其机制研究

目的:研究"生精散"对大鼠睾丸扭转复位后生精功能的影响及其机制。方法:40只雄性SD大鼠随机分为假手术组(A组),对照组(B组),生精散组小(C组)、中(D组)、大剂量组(E组),每组8只。建立左侧睾丸扭转模型,B组自扭转前1 h开始给予每日灌胃生理盐水1 ml/d,C组(0.01 g/kg.d)、D组(0.02 g/kg.d)、E组(0.03 g/kg.d)分别按体重给予灌服生精散,连续35 d后处死大鼠,对大鼠进行精液常规分析,用RT-PCR检测大鼠精子CatSper1的表达。结果:a+b级精子百分率、精子存活率、精子浓度,CatSper1基因表达,与A组[(51.30±6.60)%、(69.01±7.20)%、(40.53±7.01)×106/ml、2.04±0.77]相比,B组[(15.30±6.30)%、(44.42±6.36)%、(21.00±6.14)×106/ml、1.12±0.50),均显著降低(P<0.01);与B组相比,D组[(51.63±3.20)%、(72.09±2.20)%、(55.30±5.90)×106/ml、2.11±0.20]、E组[(55.93±3.17)%、(73.01±2.11)%、(58.33±4.90)×106/ml、2.31±0.17]均显著升高(P<0.01),而C组[(18.02±0.23)%、(48.04±7.01)%、(22.87±2.10)106/ml、1.19±0.51]升高不明显(P>0.05)。结论:生精散可以促进睾丸扭转复位后精子质量的恢复,其机制可能与调节生精功能,提高精子细胞CatSper1基因的表达有关。     

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.