Effects of chemotherapy on testicular functions

Of 26 previously normospermic patients treated with MOPP (nitrogen mustard, vincristine, procarbazine and prenisone) for Hodgkin's disease, 22 had azoospermia, 2 had severe oligozoospermia below 0.1 times 10⁶ spermatozoa/ml, 2 had 5 times 10⁶ spermatozoa/ml. Follicle stimulating hormone (FSH) increased significantly in all but 2 patients, but the luteinizing hormone (LH) increase was not significant. Prolactin, testosterone and 17β-oestradiol levels were unmodified. Chemotherapy is widely used for malignant and non-malignant diseases, the various drugs having different levels of toxicity. In order to prevent male sterility as a result of chemotherapy, the physician must choose the therapeutic agents carefully and may recommend cryopreservation of the patient's semen prior to therapy. In addition, the minimize the hazards of chemotherapy, the testis may be cooled during drug infusion and the germinal epithelium may be put to rest by desensitization through a LH releasing hormone analog.     

Quercetin mitigates fenitrothion‐induced testicular toxicity in rats

Fenitrothion (FNT) is a widely used organophosphorus pesticide in agriculture. Quercetin (QR), a plant‐derived flavonoid, has a free radical scavenging property. This study investigated the protective effect of QR on FNT‐induced testicular toxicity in rats. Twenty‐four male rats were divided into four groups. Group I (control) received normal saline. Group II was administered QR at the dose of 50 mg kg^?1 b.wt. Group III was orally administered FNT (20 mg kg^?1 b.wt). Group IV was gavaged FNT and QR together at the same doses. All administrations were performed daily by gavage and maintained for 70 days. Sperm parameters and histopathological changes in testes were investigated. Serum testosterone and luteinising hormone were estimated using radioimmunoassay kits. In testes, expressions of steroidogenic genes (3β‐hydroxysteroid dehydrogenase type 6, 17 β‐hydroxysteroid dehydrogenase type 3 and steroidogenic factor‐1) and oxidative stress genes (catalase and superoxide dismutase) were determined using real‐time PCR. FNT administration caused significant decreases in sperm count, motility and hormonal levels, a significant increase in abnormal sperm morphology and a significant down‐regulation of steroidogenic and antioxidant genes in the testis. However, QR administration ameliorated FNT‐induced toxic effects. Our results concluded that QR effectively mitigated testicular damage induced by FNT in rats.     

Recovery of testicular functions after surgical treatment of experimental cryptorchidism in the rat

When rats were made unilaterally cryptorchid at 17 days of age (before spontaneous descensus), the further maturation of the testis was prevented. At 34 days of age, the abdominal testis was smaller than the scrotal testis and showed less secretion of the Sertoli cell specific androgen binding protein (ABP). In 120–130 days old rats that were made bilaterally cryptorchid at 17 days of age, testicular weight, histology, secretion of fluid and ABP were restored and testosterone secretion and fertility were normal if orchidopexy was performed at 33 days of age. If the orchidopexy was delayed until 59 days of age, the recovery of testicular function and morphology was only partial. The results show that in the rat, the testicular damage caused by cryptorchidism is reversible, if the abdominal testis is surgically descended during early sexual maturation.     

Characteristics of the inhibitory effect of chronic treatment with an LHRH agonist on testicular steroidogenesis in the dog

Daily subcutaneous administration for 3 months of the potent LHRH agonist (D-Ser(TBU)6, des-Gly-NH2(10] LHRH ethylamide (25 micrograms) to adult dogs having spontaneous benign prostate hyperplasia (BPH) causes a marked inhibition of testicular androstenedione and testosterone secretion. This inhibition of delta 4-androgen secretion is accompanied by a decrease of testicular progestin precursors and 5 alpha-androgen metabolites, thus suggesting that, in dog, the loss of testicular steroidogenic activity, induced by the administration of an LHRH agonist, is due to a total inhibition of testicular steroidogenesis. In plasma, the concentration of both testosterone and dihydrotestosterone is also markedly depressed while androstane-3 alpha, 17 beta-diol levels remain unchanged. Measurement of prostatic steroid content has shown that administration of the LHRH agonist as well as castration is associated with a marked decrease in androstenedione, testosterone, and dihydrotestosterone levels in prostate while there is a small inhibition of androst-5-ene-3 beta, 17 beta-diol, androstane-3 beta, 17 beta-diol, dehydroepiandrosterone, and estrone concentrations in this tissue. The present data show that treatment with an LHRH agonist in the dog causes a marked inhibition of testicular steroid secretion similar to the one observed in adult men, and suggest that steroids from adrenal origin may also be involved in prostatic function.     

比较四种冷冻保护剂对大鼠睾丸组织结构的保护作用

目的 比较4种保护剂对睾丸组织结构的冷冻保存效果,探索冷冻睾丸组织的最佳保护剂.方法 选取6只雄性成龄SD大鼠(体重210～225g)的新鲜睾丸组织,分别用以下四种保护剂进行冷冻保存:甘油-卵黄-柠檬酸钠(Ⅰ组);甘油-蔗糖(Ⅱ组);丙二醇(PrOH,Ⅲ组);二甲亚砜(D M S O,Ⅳ组).光镜和透射电镜综合分析非冷冻睾丸组织和冷冻1个月后的睾丸组织的完整性,用光镜对各组进行评分,并用非参数秩和检验方法对各组之间的差异进行统计分析.结果 光镜观察,冷冻组和非冷冻组睾丸组织的生精小管、间质细胞及基质的评分差异没有统计学意义(P＞0.05),但基底膜的两组评分差异有统计学意义(P＜0.05),显示基底膜损伤较严重;用秩和检验各组保护剂对睾丸组织的冻存效果,分析结果显示各冷冻组对生精小管和间质细胞的损伤差异有统计学意义(P＜0.05),Ⅰ组和Ⅳ组的生精小管评分高于其它各组.透射电镜观察发现Ⅳ组对睾丸组织的精原细胞、支持细胞和间质细胞损伤轻微,明显优于其它各组.结论 DMSO作低温保护剂对睾丸组织的保护作用优于甘油-卵黄-柠檬酸钠、甘油-蔗糖及丙二醇.     

睾丸局部热作用对幼鼠生精细胞凋亡的影响

目的探讨睾丸局部热作用对幼鼠生精细胞凋亡的影响。方法48只健康幼年期SD雄性大鼠随机分为热处理组(43℃,15min)和对照组(22℃,15min):分别按睾丸局部处理后2h,6h,24h,14d分成4个亚组。应用电镜和原位末端标记法(TUNEL)检测各亚组幼鼠睾丸生精细胞凋亡情况,并在光镜下观察睾丸组织学变化。结果电镜显示热处理组除2h外各组均出现生精细胞凋亡表现;TUNEL法检测结果用生精细胞凋亡指数(AI)表示;热处理组2hAI与对照组相比无统计学意义,热处理组6hAI高于对照组(P<0.05),热处理组24h和14dAI明显高于对照组(P<0.01)。结论睾丸局部热作用导致幼鼠生精细胞凋亡增加。     

Quercetin has a protective role on histopathological findings on testicular ischaemia-reperfusion injury in rats

We investigated the effects of quercetin on pathological findings on testicular ischaemia-reperfusion (I/R) injury in rats. Twenty-four male Wistar albino rats were randomly assigned into four groups: Group 1, control (n = 5); Group 2, sham (n = 4); Group 3, I/R (n = 8); and Group 4, I/R + quercetin (n = 7). Bilateral testicular artery and vein were occluded for 1 h, followed by reperfusion in I/R and I/R + quercetin animals. Quercetin (20 mg kg(-1) per day) was administrated once daily by gavage to Group 1 and Group 4, respectively, after reperfusion. At the end of the study, bilateral orchiectomies were performed for histopathologic examination. The tissue damage was evaluated with light microscopy. Normal inter-stitium and seminiferous tubules were observed in control group. In the sham group, rats were seen minimal oedema around the seminiferous tubules and congested vascular structures. In Group 3, oedema, vascular congestion and haemorrhage between seminiferous tubules were observed. In Group 4, histopathologic features were markedly less than Group 3 (P = 0.03). Our study demonstrated that quercetin seems to have a protective effect on testis histopathology in rats with testicular I/R.     

Effects of vasoactive intestinal peptide (VIP) on the testicular vasculature of the rat

Using laser Doppler flowmetry, the effects of unilateral intratesticular injection of vasoactive intestinal peptide (VIP) on testicular blood flow and mean arterial blood pressure were studied in anaesthetized adult rats. At doses of 50 and 500 ng, VIP increased blood flow by 29 ± 8 and by 77 ± 25% ( p < 0.05), respectively (means ±  SEM) at 5 mm, but not at 15 mm, from the injection site. Blood pressure was not significantly affected by 50 ng, but was slightly reduced (by −8 ± 2%) after treatment with 500 ng VIP. The highest dose of VIP (5 μg) reduced blood pressure (by −30 ± 9%), and decreased blood flow at 15 mm, but not at 5 mm, from the injection site. It is concluded that VIP can act as a vasodilator in the rat testis.     

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on foetal male rat steroidogenesis

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is the most toxic and widely investigated dioxin congener. In utero and lactational exposure to TCDD results in developmental and reproductive defects that are the most sensitive endpoints for TCDD toxicity. TCDD has a potential to interfere with steroid metabolism, but the mechanisms by which this occurs are not well understood. In this study, we investigated the effects of TCDD on prenatal rat steroidogenesis. Pregnant Sprague-Dawley female rats were treated once with TCDD (0, 0.3 or 1 microg/kg) by gavage on embryonic day (ED) 11 and the expression levels of androgen (AR) and estrogen receptors (ER), steroidogenic enzymes (P450scc and 3beta-HSD) and four regulatory factors (StAR, SF-1, GATA-4 and Insl-3) involved in foetal Leydig cell and adrenal function were analysed on ED 19.5. Hormonal status of male foetuses was determined by measuring testicular testosterone (T) levels, plasma luteinizing hormone (LH) and corticosterone concentrations. In utero exposure to TCDD reduced intratesticular T of foetal males (significant at 0.3 microg/kg TCDD) and tended to reduce the protein expression of ERalpha and AR of foetal male rat testis. Foetal male rat plasma LH levels were significantly reduced at the dose of 1 microg/kg TCDD, while corticosterone levels tended to be increased possibly because of the TCDD-induced stress. Only minor alterations in steroidogenesis were observed in rat adrenal. mRNA expression of developmental regulatory factors was not influenced by foetal TCDD exposure, except for significantly reduced adrenal SF-1. The results demonstrate that maternal exposure to TCDD suppressed testicular steroidogenesis of 19.5-day-old foetal male Sprague-Dawley rat. The highest dose of TCDD (1 microg/kg) had also an effect on pituitary LH secretion. Our data implicate that TCDD has direct testicular and pituitary effects on foetal male rat but with different dose-responses. These changes can lead to impaired steroidogenesis and it may result in the maldevelopment of the testis and weaken masculinization.     

Protective effect of Zingiber officinale extract on rat testis after cyclophosphamide treatment

Decreasing the side effects of chemotherapy in testis has been the subjects of many studies. In this study, the protective effects of Zingiber officinale extract on rat testis were investigated after chemotherapy with cyclophosphamide. Histological and biochemical parameters were compared in cyclophosphamide‐treated rats with or without ginger extract intake. Wistar male rats were randomly divided into four groups each 10. The control group received a single injection of 1 ml isotonic saline intraperitoneally. The Cyclophosphamide (CP) group received a single dose of cyclophosphamide (100 mg kg^?1 BW) intraperitoneally. CP + 300 and CP + 600 groups received orally 300 or 600 mg of ginger extract, respectively, for a period of 6 weeks after cyclophosphamide injection. The morphologic and histological structure of the testis was compared in different groups of the rats. Also, factors like malondialdehyde, reactive oxygen species, total antioxidant capacity and testosterone level were assessed in blood serum as well. Our results showed that although ginger extract could not change testis weight, malondialdehyde (MDA) and ROS, but antioxidant and testosterone levels in serum were increased significantly. Also, an obvious improved histological change was seen in CP + 300 and CP + 600 groups in comparison with CP group. These protective effects of ginger on rat testis after cyclophosphamide treatment could be attributed to the higher serum level of antioxidants.     

The effect of poly (adenosine diphosphate-ribose) polymerase inhibitors on biochemical changes in testicular ischemia-reperfusion injury

PURPOSE: Poly (adenosine diphosphate [ADP]-ribose) polymerase inhibitors have been used successfully to decrease ischemia-reperfusion injury in several organ systems. We evaluated the efficacy of poly (ADP-ribose) polymerase inhibitors on biochemical changes in testicular ischemia-reperfusion injury. MATERIALS AND METHODS: Adult male Wistar rats were divided into 9 groups of 6 each. One group served to determine baseline values of biochemical parameters, 1 that underwent sham operation served as a control, 1 underwent 2 hours of testicular torsion and 4 hours of detorsion, 2 received pretreatment with vehicle (saline or dimethyl sulfoxide) before detorsion and 4 received pretreatment with the poly (ADP-ribose) polymerase inhibitor nicotinamide, 3-aminobenzamide, 1,5-dihydroxyisoquinoline or 4-amino-1,8-naphthalimide before detorsion. Lipid peroxidation products, nitric oxide content and myeloperoxidase activity, an indicator of neutrophil accumulation, were assessed in testicular and renal tissues. RESULTS: Testicular torsion-detorsion caused a significant increase in lipid peroxidation products, nitric oxide content and myeloperoxidase activity in ipsilateral testes (p <0.01) but not in the contralateral testes or kidneys. Animals treated with poly (ADP-ribose) polymerase inhibitors had a significant decrease in these biochemical parameters compared with vehicle treated animals (p <0.01). CONCLUSIONS: These data emphasize that poly (ADP-ribose) polymerase may have a role in testicular damage caused by ischemia-reperfusion and the inhibition of poly (ADP-ribose) polymerase may be a novel approach to therapy for ischemia-reperfusion injury of the testis.     

Age-specific effect of phthalate ester on testicular development in rats

Purpose

Phthalate esters have been shown to induce testicular damage in both adult and immature rats; however, there have been, so far, few reports describing the age-specific effects of phthalate esters on testicular function. The aim of this study is therefore to investigate the age-specific effects of mono-n-butyl phthalate (MBP) on the testes in both prepubertal and mature adult rats.

Materials and methods

Both prepubertal male rats and adult mature male rats were fed special rat chow containing 1% MBP for 10 days. Control prepubertal and adult rats were fed standard commercial rat chow during the same period as the MBP-treated rats. After 10 days of feeding, all rats were killed, and the testes were removed. The weight of the testis was measured, and histological examination of the testis was performed. In addition, the frequency of an apoptotic cell appearance in the seminiferous tubules was determined in both MBP-treated and control groups.

Results

In the prepubertal rats, the mean weight of the testes was significantly lower in the MBP-treated rats than in the control rats. A histological examination of the MBP-treated testes showed a decreased seminiferous tubular diameter and an inhibited maturation of germ cells in comparison to those of the control testes. Furthermore, apoptotic cells appeared more frequently in the MBP-treated testes than in the control testes. Although in adult mature rats, no significant difference was observed in either the testicular weight or the histological findings between the MBP-treated and control rats.

Conclusions

The oral administration of MBP to male rats was observed to produce more pronounced testicular damage in prepubertal rats than in adult mature rats. Immature testes may thus be more sensitive to MBP, which induces the germ cell apoptosis in seminiferous tubules and testicular atrophy in prepubertal young rats.     

糖尿病对小鼠睾丸组织中PCNA表达的影响

为研究糖尿病对睾丸生殖细胞DNA合成的影响，应用免疫组化ABC法检测增殖细胞核抗原（proliferation cell nuleus antigen PCNA)在睾丸组织的表达。结果显示免疫阳性细胞包括精原细胞和初级精母细胞，少数支持细胞和肌样细胞胞核也有着色。PCNA阳性率随糖尿病病程进展呈明显下降趋势，而且每年龄段糖尿病组PCNA阳性率均明显低于相应正常对照组。由此得出结论，糖尿病对PCNA在睾丸组织中表达起抑制作用，且与糖尿病病程呈正相关，即糖尿病时生精过程受到明显抑制，这种抑制作用可能与糖尿病所导致的雄性生殖能力下降有关。     

The effect of varicocele repair on testicular volume in children and adolescents with varicocele

PURPOSE: We investigated the effect of varicocele repair on testicular volume according to age in children and adolescents and review the long-term results of varicocele surgery. MATERIALS AND METHODS: The study included 39 boys 11 to 19 years old with clinical palpable varicocele who underwent varicocele surgery with at least 1 year of postoperative followup. Preoperative and postoperative testicular volumes were monitored and measured with an ellipsoid Prader orchidometer. Physical examination findings (testicular volumes and testicular consistency) in all boys, and serum hormone values and semen parameters in 16 adolescents were recorded and compared before and after surgery. RESULTS: Left unilateral varicocelectomy was done in 29 boys (74%) and bilateral varicocelectomy in 10 (26%). While no postoperative hematoma, infection or testicular atrophy was observed, 1 boy (2.5%) had varicocele recurrence and 2 boys (5.1%) had minimal hydroceles that required no intervention. Significant increases were observed in postoperative sperm concentration (p = 0.01), total motile sperm count (p = 0.009), testis volume (p = 0.000) and serum testosterone level (p = 0.014). All 15 boys with preoperative soft testis had normal testicular consistency postoperatively. Of the 19 boys with preoperative testicular atrophy 10 (53%) did regain normal testicular growth, while 9 (47%) retained testicular volume loss after surgery. When comparing preoperative to postoperative increase in testicular volume according to age in all boys, the mean was statistically significantly higher in boys younger than 14 years (left testis p = 0.037, right testis p = 0.000). CONCLUSIONS: Testicular consistency achieved normal firmness after varicocelectomy in all boys with preoperative soft testis. While there was catch-up growth in comparison to the contralateral testis, testicular consistency improved but testicular volumes may not increase significantly after varicocele repair at ages older than 14 years. However, in these adolescents postoperative semen parameters and serum hormone values may significantly improve regardless of testicular volume. Therefore, boys with varicocele and their families should be fully informed in light of these findings.     

Protective effect of ebselen on experimental testicular torsion and detorsion injury

Ebselen is used as a drug in clinical trials against stroke, reperfusion injury with anti‐atherosclerotic and renoprotective effects. The aim of this study is to investigate the protective effect of ebselen, on torsion/detorsion (T/D)‐induced biochemical and histopathological changes in experimental testicular ischaemia/reperfusion injury. A total of 28 male Wistar Albino rats were divided into four groups: group 1(sham‐operated group, n = 7), group 2(ebselen group, n = 7), group 3(torsion/detorsion + saline, n = 7) and group 4(T/D + 10 mg kg^?1 ebselen group, n = 7). The tissue homogenate samples were used for immediate nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase, catalase and glutathione measurement. Testes in all groups were evaluated for the biochemical assay and histopathological examinations. To evaluate spermatogenesis, Johnsen scoring system was used. Testicular tissue MDA and NO levels in group 3 were significantly higher than in group 1 and 4. In histological evaluation of the testicular tissues, ebselen administration improved tubular histology significantly compared with T/D group. Significant increase in histological score was observed in the testis of group 3 compared with group 1 and 2. Histological score in group 4 significantly decreased compared with group 3. Johnson score was significantly lower in T/D group compared with all other three groups, ebselen administration increased the score significantly compared with T/D group. Ebselen reduced oxidative biochemical and histopathological damage in our testicular T/D rat model.     

Ciliated Cells in the Rat Testis Effects of Neonatal Administration of Estrogens

The presence of cells bearing solitary cilia has been studied in the testes of normal and neonatally estrogenized rats. Peritubular myoid cells and undifferentiated interstitial cells showed a higher frequency of cilia in estrogenized animals than in control ones, from 15 to 45 days of age, the differences being slighter at 90 days of age.     

Influence of rat testicular macrophages on Leydig cell function in vitro

The influence of co-cultures of rat testicular macrophages and Leydig cells (LC) on LC morphology and steroidogenesis was investigated with and without macrophage stimulation by a bacterial lipopolysaccharide (LPS). LC showed an elongated form in the presence of stimulated testicular macrophages. In the presence of non-stimulated testicular macrophages a significant inhibition of testosterone production was observed (decrease of 33%) from 48 h in co-culture while an increase of 16% was obtained at the same culture time, after stimulation of macrophages by LPS. When LC were treated with testicular macrophage-conditioned media (MCM) obtained from LPS-treated macrophages, they became fusiform and there was stimulation (78%) of steroid production. After human FSH stimulation (1-1000 mIU ml-1), MCM from testicular macrophages was no more effective in enhancing testosterone production by LC than was media from untreated LC. Similar experiments with LPS were conducted with macrophages of peritoneal origin. Peritoneal macrophages stimulated or not by LPS in co-cultures with LC or peritoneal MCM did not significantly modify testosterone production. However, these cells were able to modify LC morphology when LPS-MCM was added to LC-culture medium. The present results suggest strongly that testicular macrophage-LC interactions could be important in the control of LC steroidogenesis.     

Studies on relationship between testicular capsule and sperm transport in rat testis

Aim: In SD rats, histological changes in the testis were observed after bilateral capsulotomy (of the tunica albuginea) in order to investigate the physiological role of the testicular capsule on sperm transport. Methods: Bilateral longitudinal capsulotomy was devised to disrupt the capsular contractile function. With this technique, only the tunica vaginalis and tunlca albuginea were slit open, leaving the tunica vasculosa intact to embrace the underlying testicular parenchyma. After capsulotomy, the structural changes in the seminiferous tubules, the transitional distal seminiferous segment, and the rete testis were observed. Results: In the capsulotomized testis, there was sperm retention at the transitional seminiferous segment and progressive degenerative changes in seminiferous tubules. Conclusion: The results clearly indicated that an intact testicular capsule was required for normal sperm transport from the seminiferous tubules into the rete testis. This is the first attempt to study the physiological role of the testicular capsule in intact animals.     

Effects of various cryopreservation media and freezing–thawing on the morphology of rat testicular biopsies

Currently, testicular sperm extraction is successfully combined with intracytoplasmic sperm injection into the oocyte (ICSI). Several pieces of a testicular biopsy can be frozen and thawed until the ICSI attempt. In this study, the effects of freezing-thawing on the morphology of rat testicular biopsies stored in different cryopreservation media were analysed. Each cryopreservation medium contained glycerol and/or dimethyl sulfoxide (DMSO) as cryoprotectants. In general, both glycerol and DMSO, when applied at moderate concentrations (6-25%), preserved the structure of the seminiferous epithelium. The freezing-thawing procedure had no significant effect on tubular diameter; however, it caused a 'folding' of the lamina propria and notable damage to Sertoli cells, spermatogonia and spermatocytes. Round and elongated spermatids and spermatozoa displayed occasional nuclear damage, vacuolization, and shrinkage/swelling of the cytoplasm. However, the vast majority of these cells maintained their normal structure in nearly all the applied cryomedia. It is concluded that freezing-thawing of testicular biopsies, and the cryopreservation medium, have a significant impact on the structure of the seminiferous epithelium, particularly on its basal compartment.