Valproic acid changes the expression of tyrosine‐phosphorylated proteins in rat seminal vesicle

Previous studies reported the effects of valproic acid (VPA) on tyrosine‐phosphorylated (TyrPho) protein expression in the testis and epididymis, but its effects on that in the seminal vesicle (SV) have never been demonstrated. This study attempted to investigate the expressions of TyrPho proteins in SV treated with VPA. Sixteen rats were divided into control and VPA‐treated groups. The control rats were injected with normal saline, whereas the treated animals were intraperitoneally injected with VPA (500 mg/kg BW) for 10 consecutive days (n = 8/each). The biochemical parameters in blood plasma and SV fluid were analysed. The SV tissues and fluid were investigated for the presence and expression of TyrPho proteins, androgen receptor (AR) proteins and heat shock protein 70 (Hsp70). Significantly, VPA caused SV atrophy and reductions in secretion and biochemical parameter levels. There were significant increases in many TyrPho proteins in the plasma (a 95 kDa) and SV tissue (a 40 kDa) of the VPA rats. However, the expressions of seminal AR, Hsp70 and TyrPho proteins (50 and 48 kDa) were significantly lower in VPA rats. Recent results have indicated that VPA affected SV morphology and decreased biochemical fluid substances including TyrPho proteins associated with decreased expressions of AR and Hsp70.     

Chronic stress affects tyrosine phosphorylated protein expression and secretion of male rat epididymis

Chronic stress (CS) is shown to decrease the semen quality with changed expression of tyrosine phosphorylated (TyrPho) proteins in testicular and seminal tissues. However, the alterations of such proteins and fluid contents in the epididymis, producing sperm maturation factors, have never been reported. Sixteen adult rats were randomly divided into 2 groups (n = 8). The control animals were not subjected to stressors whereas CS rats were immobilised within restraint cage (4 hr/day) before cold forced-water swimming (15 min/day) for 60 days. Corticosterone, testosterone, blood glucose level (BGL), malondialdehyde (MDA) and biochemical components in epididymal fluid were assayed. Expressions of heat shock protein 70 (HSP-70), androgen receptor (AR) and TyrPho protein were investigated in epididymal tissue and fluid. Significantly, CS increased the corticosterone and BGL but decreased testosterone and epididymal substance levels. MDA level in tail epididymal fluid and HSP-70 expression in both regions of epididymal tissues and fluids, except in head epididymal fluid of CS were increased. Epididymal tissues showed the decrease of AR expression. Presence and changes of many TyrPho proteins were observed in CS. In conclusion, CS could affect functional proteins particularly TyrPho in epididymis, resulted in low semen quality.     

Opposite effects of unmodified prolactin and a molecular mimic of phosphorylated prolactin on morphology and the expression of prostate specific genes in the normal rat prostate

BACKGROUND: In the current study, we have investigated the individual roles of unmodified, wild-type prolactin (WT PRL) and a molecular mimic of phosphorylated prolactin (S179D PRL) in the normal rat prostate. METHODS: In the first animal experiment, recombinant WT PRL and S179D PRL were delivered to adult male rats at a rate of 14 microg/kg per day for 3 weeks. In the second animal experiment, two subcutaneous (200 microg/kg) injections of long-acting forms of the two PRLs were given to adult male rats on day 1 and day 22 for a total of 5.5 weeks of treatment. RESULTS: The different forms of PRL had opposite effects on the normal rat prostate, independently of androgens. WT PRL promoted morphologic changes in prostate epithelium consistent with preparation for cell proliferation, whereas S179D PRL produced morphologic evidence of a more differentiated epithelium. Northern blot analysis of expression of the two major prostate specific proteins, prostatein and probasin, showed that WT PRL decreased, whereas S179D PRL increased, the expression of the mRNAs for these two proteins. At the same time, S179D PRL reduced both testosterone and dihydrotestosterone levels. CONCLUSION: We conclude that PRL is an important modulator of normal rat prostate biology and that different forms of PRL have specific functions. The molecular mimic of phosphorylated PRL, S179D PRL, is the most important in terms of epithelial cell differentiation.     

氯沙坦对自发性2型糖尿病KKAy小鼠肾小球蛋白表达谱的影响

目的 利用比较蛋白质组学双向电泳技术研究体系来观察血管紧张素受体拮抗剂(ARB)氯沙坦对自发性2型糖尿病KKAy小鼠肾小球蛋白表达谱的影响.方法 8周龄自发性2型糖尿病KKAy小鼠随机分为氯沙坦治疗组(饮用水中喂入氯沙坦粉末10 mg+kg-1·d-1)和非治疗组;8周龄C57BL/6小鼠作为正常对照组.饲养12周后,经胸主动脉磁珠灌流分离肾小球,提取肾小球蛋白.DIGE最小荧光标记法标记,行二维荧光差异凝胶电泳.应用Typhoon多功能成像系统扫描凝胶,DeCyder 2D差异分析软件进行图像分析.采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)鉴定差异表达蛋白质.结果 KKAy小鼠的体质量、血糖和尿白蛋白排泄率较正常对照C57BL/6小鼠显著升高(均P＜0.05),氯沙坦治疗显著改善2型糖尿病KKAy小鼠尿白蛋白肌酐比[(539.71±100.23)mg/g比(728±177.19) mg/g]、肾小球基底膜增厚和系膜基质增生等肾脏病理损害,而对血糖无影响.通过DeCyder 2D差异分析软件,发现KKAy氯沙坦治疗组与KKAy非治疗组肾小球内表达差异大于1.1倍的蛋白质斑点62个.经肽质量指纹图分析,鉴定出41种蛋白质.其中表达上调的蛋白28种,包括甘油激酶、亚硫酸盐氧化酶、甘氨酸脒基转移酶、腺苷高半胱氨酸酶等;表达下调的蛋白质13种,包括3-巯基丙酮酸硫基转移酶、ATP合酶亚单位d、60 000热休克蛋白、线粒体应激蛋白70(又名75 000葡萄糖调节蛋白,GRP75)等.有6种蛋白在20周龄KKAy小鼠和C57BL/6小鼠肾小球内存在差异表达,氯沙坦治疗抑制了其在糖尿病状态下的表达变化,包括丙酮酸脱氢酶复合物的二氢硫辛酰赖氨酸残基乙酰基转移酶组分、琥珀酰辅酶A连接酶[GDP-形成]亚单位β、ATP合酶亚单位d、GRP75、核苷二磷酸盐结合部分X模体19和硒结合蛋白1.结论 氯沙坦可明显减轻KKAy小鼠尿白蛋白排泄率和肾脏病理损害;可抑制糖尿病状态下肾小球ATP合酶亚单位d、GRP75、硒结合蛋白1等蛋白的表达变化;可能通过减少线粒体活性氧簇产生,抑制氧化应激反应发挥其肾脏保护作用.     

Therapeutic effect of pentoxifylline on reproductive parameters in diabetic male mice

In this study, we aimed to investigate the effects of pentoxifylline (PTX) on male reproductive parameters in diabetic mice. Male adult mice (n = 24) were divided into control and three experimental groups (n = 6) including Diabetic, Diabetic + PTX and PTX groups. Diabetes was induced by single injection of streptozotocin (60 mg kg^?1). PTX was administered intraperitoneally at the dose of 12 mg kg^?1 for 14 days 1 week after diabetes induction. Serum levels of testosterone and blood glucose were determined and collected spermatozoa from cauda epididymidis analysed. Based on histological slides prepared from testis, the diameter of seminiferous tubules was determined using Motic camera and software and also apoptosis using TUNEL assay. Data were analysed using one‐way anova method, and P < 0.05 was considered statistically significant. The mean of seminiferous tubules diameter, final body weight, testis weight, sperm parameters and testosterone hormone level in PTX‐treated diabetic group indicated a significant increase compared to diabetic one, whereas apoptosis index and blood glucose were decreased in this comparison (P < 0.05). These results suggest that intraperitoneal administration of PTX is a potentially beneficial agent to reduce testicular damage and improves sperm parameters in diabetic mice by decreasing the ratio of apoptosis.     

Terazosin‐induced alterations in catalase expression and lipid peroxidation in the rat seminal vesicles

Previous studies have shown that alpha1‐adrenergic receptor antagonists may alter seminal vesicle contractility and impair fertility in male rats. This study was designed to investigate the effects of terazosin on the catalase expression in the seminal vesicles and the lipid peroxidation of the seminal fluid in normal adult rats. Wistar rats were treated with terazosin (1.2 mg kg^?1 body weight, given orally every second day) for 120 days. Catalase expression was assessed immunohistochemically in tissue sections of the seminal vesicles, and lipid peroxidation was estimated by measuring the malondialdehyde (MDA) levels in the seminal vesicles' fluid. The seminal vesicles in terazosin‐treated rats were particularly distended in comparison with those of controls, and their secreting epithelium was suppressed. Cytoplasmic catalase expression in the secreting epithelial cells (% of cells) was increased in terazosin‐treated specimens in comparison with controls (76.1 ± 17.1 versus 51.3 ± 25.1, P = 0.005). MDA levels (μm ) were also higher in samples from treated subjects in comparison with controls (2.67 ± 1.19 versus 1.39 ± 0.19, P = 0.01). Although the direct effect of terazosin treatment on the seminal vesicles is that of impaired contractility, an indirect effect is that on fertility by increasing lipid peroxidation in the seminal fluid and/or through degrading of hydrogen peroxide that is essential for sperm capacitation.     

Melatonin prevents deterioration of erectile function in streptozotocin-induced diabetic rats via sirtuin-1 expression

A review of the literature indicated that sirtuin-1 expression, a regulator of nitric oxide bioavailability in erectile dysfunction (ED) after melatonin therapy, has not yet been investigated. The objective of this study was to evaluate the protective effects of melatonin for erectile function with sirtuin-1 protein expression in type 1 diabetic rat models. Fifty male Sprague Dawley rats were placed into five groups. Except for those in the control group (C), each animal received a single dose (60 mg/kg) of streptozotocin to induce diabetes. The animals were placed into the diabetes (D) group, insulin (I) group (6 U/kg/day), melatonin (Mel) group (10 mg kg⁻¹ day⁻¹) and combined treatment (I + Mel) group. Ten weeks later, the serum testosterone levels, intracavernosal pressure (ICP), mean arterial pressure (MAP), malondialdehyde (MDA), cyclic guanosine monophosphate (c-GMP), 8-hydroxydeoxyguanosine (8-OHdG), nitric oxide synthase (NOS), caspase-3 activity, sirtuin-1 and endothelial nitric oxide synthase (eNOS) protein expression and histological findings were assessed. The mean ICP/MAP ratio for the D group was lower than the mean ratios for the other groups. The treatment groups, particularly the I + Mel group, exhibited lower 8-OHdG and MDA levels and caspase-3 activity than the D group. The sirtuin-1 and eNOS expression and cavernosal tissue (CT) histology seemed to have been preserved by the melatonin and/or insulin therapy. These results were indicative of a profound protective effect of melatonin by the activation of sirtuin-1 protein expression against hyperglycemia-induced oxidative CT injury.     

Additional deleterious effects of alcohol consumption on sperm parameters and DNA integrity in diabetic mice

The aim of this study was to survey the impact of alcohol consumption on sperm parameters and DNA integrity in experimentally induced diabetic mice. A total of 32 adult male mice were divided into four groups: mice of group 1 served as control fed on basal diet, group 2 received streptozotocin (STZ) (200 mg kg^?1, single dose, intraperitoneal) and basal diet, group 3 received alcohol (10 mg kg^?1, water soluble) and basal diet, and group 4 received STZ and alcohol for 35 days. The cauda epididymidis of each mouse was dissected and placed in 1 ml of pre‐warm Ham's F10 culture medium for 30 min. The swim‐out spermatozoa were analysed for count, motility, morphology and viability. Sperm chromatin quality was evaluated with aniline blue, toluidine blue, acridine orange and chromomycin A3 staining. The results showed that all sperm parameters had significant differences (P < 0.05), also when sperm chromatin was assessed with cytochemical tests. There were significant differences (P < 0.001) between the groups. According to our results, alcohol and diabetes can cause abnormalities in sperm parameters and chromatin quality. In addition, alcohol consumption in diabetic mice can intensify sperm chromatin/DNA damage.     

Berberine modulates crucial erectogenic biomolecules and alters histological architecture in penile tissues of diabetic rats

Berberine is an isoquinoline alkaloid, found in several plants. Diabetes induces erectile dysfunction (ED) via reduction in some hormones and enzymes implicated in sexual function. This study aimed to investigate the role of berberine on crucial biomolecules linked to penile function in diabetic rats. Sixty-three (63) adult male rats were used and distributed into nine groups (each = 7). Group I–IV normal rats administered with citrate buffer (pH 4.5), sildenafil citrate (SD, 5.0 mg/kg), 50 and 100 mg/kg of berberine, respectively, via oral gavage. Rats in groups V–IX were diabetic rat with ED treated with buffer, SD, 50 and 100 mg/kg of berberine, and acarbose (25 mg/kg ACA) respectively. The result revealed that histological architecture in penile tissues were altered in diabetic groups treated with berberine, sildenafil citrate and acarbose when compared to the diabetic control group. Treatment with berberine, increased testosterone, luteinizing hormone and follicle-stimulating hormone in diabetic rat with ED. Also, reduced prolactin level and acetylcholinesterase, angiotensin-1 converting enzyme, adenosine deaminase and arginase activities were observed in berberine treated diabetic rat with ED. Molecular docking analysis revealed that berberine had strong binding affinities for these enzymes. Thus, berberine could represent a potential therapeutic agent for diabetes-induced ED.     

The use of steroid-containing silastic implants in male nude mice: Plasma hormone levels and the effect of implantation on the weights of the ventral prostate and seminal vesicles

The effects of implantation of steroid-containing capsules into male nude mice on steroid concentrations in plasma and weight of accessory sex organs were studied. Intact male nude mice had plasma levels of testosterone (T) of 8.0 ± 2.4 ng/ml, while T implantation (length 1.0 cm) of a group with castrated male mice resulted in a mean level of 8.1 ± 0.3 ng/ml. This level was reached within 2 days after implantation and lasted for at least 40 days. After longer periods of application (up to 75 days) physiological levels could still be attained. Treatment of intact male nude mice with estradiol (E2)-containing implants (length 0.5 cm) resulted in constant levels of plasma-E2 (250 pg/ml) also lasting for at least 32 days. This treatment resulted in a mean plasma-T level of 1.0 ± 0.2 ng/ml, which was still significantly higher than that obtained after castration (0.14 ± 0.03 ng/ml). Up to 16 days after implantation E2 did not cause a decrease of the weights of the accessory sex glands in intact male mice, while after 32 days a significant reduction (50% of the control animals) of the organ weights was observed. The present data obtained with T and E2 implantation show that this route of administration of hormones is also very applicable in the nude mouse model.     

慢性前列腺炎患者精浆中热休克蛋白的表达及临床意义

目的　探讨慢性前列腺炎患者精浆中热休克蛋白 70 (HSP 70 )的表达及临床意义。　方法　采用酶联免疫吸附测定法 (ELISA)对 10例慢性细菌性前列腺炎 (Ⅱ型 )、10例炎症型慢性非细菌性前列腺炎即炎症型慢性骨盆疼痛综合征 (ⅢA 型 )、10例非炎症型慢性非细菌性前列腺炎即非炎症型慢性骨盆疼痛综合征 (ⅢB 型 )患者及 6例健康志愿者精浆中HSP 70含量进行测定 ,并与慢性前列腺炎症状评分 (修订版 )进行相关性研究。　结果　Ⅱ型、ⅢA 型、ⅢB 型慢性前列腺炎患者及正常对照组精浆HSP 70含量分别为 (10 .74± 4 .0 2 )ng/ml、(2 .5 8± 0 .96 )ng/ml、(2 .78± 1.12 )ng/ml及 (6 .5 7±2 .72 )ng/ml。Ⅱ型患者精浆HSP 70含量显著高于正常对照组 ,P <0 .0 1,差异有显著性意义。ⅢA 型与ⅢB 型患者HSP 70含量均显著低于正常对照组 ,P <0 .0 1,差异有显著性意义。而ⅢA 型与ⅢB 型患者HSP 70含量间差异无显著性意义 ,P >0 .0 5。各组精浆HSP 70含量与慢性前列腺炎症状评分间均呈显著的负相关关系 (P <0 .0 1)。　结论　测定精浆中HSP 70的表达对各种类型慢性前列腺炎诊断有一定的临床应用价值 ,并可作为评价慢性前列腺炎病情程度的分子生物学指标。HSP 70在慢性前列腺炎的治疗方面也具有广阔的应用前景。     

链脲佐菌素诱导的糖尿病模型小鼠对炎症性关节炎的影响

目的 探讨链脲佐菌素(STZ)诱导的Ⅰ型糖尿病模型对小鼠炎症性关节炎的影响.方法 将16只6周龄SPF级雄性C57BL/6J小鼠随机分成两组,每组8只.糖尿病组小鼠采用于6周龄时一次性腹腔注射链脲佐菌素150 mg/kg建立小鼠Ⅰ型糖尿病模型;对照组注射同等体积柠檬酸盐缓冲液.在7周龄时对两组小鼠均于双侧后足足垫注射牛...     

SHG-44绿色荧光裸鼠原位模型的建立

目的 建立SHG-44绿色荧光裸鼠原位模型.方法 将C57BL/6J-增强绿色荧光蛋白(ECFP)转基因小鼠与裸小鼠进行交配,在继代时严格挑选都表达EGFP基因的无毛雄鼠(♂)与有毛母鼠(♀)交配,通过肉眼、荧光显微镜和分子生物学等方法观察EGFP基因在裸小鼠全身各组织中的表达;将胶质瘤细胞株SHG-44(15μl,含1×106个细胞)注射于绿色荧光裸鼠的脑内制成原位模型.结果 每只种鼠平均产仔7～8只,其中纯合裸仔鼠有3～4只,比例接近1∶1;每代仔鼠通过小动物活体成像系统出生后2d就可鉴别其是否表达EGFP基因;器官及组织冰冻切片荧光显微镜下观察均见有绿色荧光;聚合酶链反应(PCR)结果表明脑组织、皮下肌肉、鼠尾和脚趾全部表达EGFP基因;原位模型的成瘤率达100％,瘤细胞依然具有人脑胶质瘤的基本特征.结论 表达EGFP基因的裸小鼠可用于肿瘤移植实验.     

Effect of androgen deprivation on the expression of aquaporins in rat prostate and seminal vesicles

The aim of this study was to investigate the level of secretions of prostate and seminal vesicles and its association with the expression of AQP0, 1, 4, 5, 6 and 8 in castrated rats. Eight‐week‐old male Sprague‐Dawley (SD) rats (n = 18) were randomly divided into control group, castrated rats group and castrated followed testosterone replacement group. Four weeks after surgery, the secretions and expression of AQP0, 1, 4, 5, 6 and 8 of prostate and seminal vesicles were determined. Serum testosterone was significantly lower in castrated groups than in control and testosterone replacement groups (P < 0.05). The level of prostate and seminal vesicle secretions and the expressions of AQP0, 1, 4, 5, 6 and 8 in prostate and seminal vesicles were significantly lower in castrated group than in control and castrated followed testosterone replacement groups (P < 0.05). The decreased prostatic and seminal vesicle secretions in castrated rats may be related to the decrease in AQP0, 1, 4, 5, 6 and 8 in prostatic tissue and seminal vesicles.     

热休克蛋白90促进糖尿病创面愈合的作用研究

目的：研究HSP90在糖尿病性难愈创面中的作用。方法：在链脲佐菌素诱导的糖尿病大鼠背部做d=2cm的全层皮肤缺损模型,并随机分为四组：空白组（B组）、HSP90组（C组）、胰岛素组（D组）及HSP90与胰岛素联合用药组（E组）。另设一组正常大鼠同样在背部做d=2cm全层皮肤缺损模型作为对照组（A组）,每组n=（5n1＋n2）=20只,共100只大鼠,在创面形成后即时给予难愈创面药物治疗持续2天,A组则不治疗。于创面形成后第3天、第5天、第7天、第10天、第14天及第21天进行大体观察及活体取材做常规病理学观察,各时间点标本采用免疫组化ABC法观察HSP90,并计算其平均密度值及积分光密度值,观察其表达规律。结果：大体观察下,在计算第21天的创面愈合率后我们发现,C组（85.9%）、D组（86.8%）及E组（96.8%）相较于B组（79.7%）创面愈合率均有显著提高（P〈0.05）,其中E组疗效更加突出;而在镜下观察中,我们发现E组创面表皮增厚,上皮化活跃,表皮细胞排列规则,且HSP90在各时期的表达水平显著增高（P〈0.05）,C组与D组虽也有类似改变但不如E组显著。结论：HSP90能够提高糖尿病难愈性创面21天的愈合率,同时我们发现将HSP90与胰岛素联合应用后会取得更好效果,进一步提高创面21天时的愈合率。     

Measurement of protein C inhibitor in seminal plasma is useful for detecting agenesis of seminal vesicles or the vas deferens

Protein C inhibitor (PCI), a plasma serine protease inhibitor of activated protein C, is present at high concentrations in the seminal plasma of normal subjects and is decreased in some infertile patients. We measured the concentrations of PCI, prostate-specific antigen, and fructose in the seminal plasma of infertile patients (n = 125) and of normal subjects (n = 13). We also measured time-dependent changes in the concentrations of PCI and fructose in seminal plasma after ejaculation. A weak correlation was found between the levels of PCI and fructose (r = 0.268, P = 0.016). The PCI level in seminal plasma of patients with seminal vesicle and/or vasal agenesis was significantly lower (P < .01) than in normal subjects. The level of fructose in seminal plasma decreased in vitro in a time-dependent manner after ejaculation, whereas the concentration of PCI was stable at 48 hours after ejaculation. These data suggest that PCI in seminal plasma, as well as fructose, may become one of the markers for agenesis of seminal vesicles and/or the vas deferens.