脂多糖对雄性大鼠睾丸组织病理学和生殖内分泌功能的影响

目的:研究脂多糖诱导的炎症对雄性大鼠睾丸组织病理学和生殖内分泌功能的影响,初步探讨炎症影响雄性生育的可能机制。方法:36只雄性SD大鼠(400~450g)随机分为4组,每组9只。对照组(A组)大鼠腹膜内注射无菌生理盐水,余下3组大鼠腹膜内注射脂多糖(LPS,5 mg/kg,溶于无菌生理盐水中),分别于注药12 h(B组)、24 h(C组)和72 h(D组)后麻醉处死。取各组大鼠左侧睾丸,制成组织切片,HE染色,镜下观察睾丸组织病理学改变;取各组大鼠血清进行血清睾酮(T)、卵泡刺激素(FSH)和黄体生成素(LH)含量检测。结果:①大鼠睾丸组织HE染色显示:与A组相比,B组大鼠睾丸生精小管结构清晰,各级生精细胞排列整齐,部分生精小管管腔内精子数目有所减少,并可见脱落的生精细胞;C组大鼠睾丸生精上皮变薄,生精小管结构较紊乱、不规则,各级生精细胞减少且排列不整齐,生精小管管腔内成熟精子数目减少,并可见脱落的生精细胞;D组大鼠睾丸生精上皮变薄,生精小管结构紊乱、不规则,各级生精细胞明显减少且层次紊乱,生精小管管腔内成熟精子数目减少,可见较多生精细胞脱落并阻塞管腔。②各组大鼠血清T、LH、FSH含量,A组T为(0.490±0.028)ng/ml,LH为(6.290±0.515)ng/L,FSH为(1.837±0.127)IU/L;B组T为(0.460±0.024)ng/ml,LH为(5.881±0.124)ng/L,FSH为(1.707±0.098)IU/L;C组T为(0.417±0.021)ng/ml,LH为(5.123±0.271)ng/L,FSH为(1.620±0.115)IU/L;D组T为(0.378±0.021)ng/ml,LH为(4.504±0.279)ng/L,FSH为(1.562±0.216)IU/L;与A组相比,B组大鼠血清T、LH和FSH含量均降低,但无统计学差异(P0.05);C、D组大鼠血清T和LH明显降低(P0.01),FSH降低,有统计学差异(P0.05)。结论:脂多糖(5 mg/kg)腹膜内注射诱导炎症可损害雄性大鼠睾丸组织,并影响大鼠生殖内分泌功能,导致血清T、LH和FSH含量降低,可能影响雄性生育。     

二甲磺酸乙烷致间质细胞破坏所引起的成年大鼠睾丸和附睾的组织学变化:形态定量研究

目的:定量研究睾酮分泌剧烈减少所致睾丸和附睾的组织学变化。方法:14只成年 SD 大鼠腹腔内注射二甲磺酸乙烷(EDS,75mg/kg),14只注射生理盐水作为对照。7天后处死各组中的一半动物,过5天后处死另一半。取睾丸和附睾组织块,甲基丙烯酸树脂包埋。用体视学的光学体视框技术估计睾丸内的细胞数,并用其它形态定量研究方法获取另外一些参数。结果:EDS 注射使睾丸内的间质细胞几乎完全消失,但对支持细胞总数没有影响。EDS 注射7天后,生精上皮内可见许多长形精子细胞滞留,附睾管内可见许多圆形精子细胞。EDS 注射12天后,精子细胞和精母细胞的排列明显变疏松,生精细胞之间出现明显的裂隙,裂隙近似放射状朝向生精小管腔;睾丸内的非 B 型精原细胞总数和精母细胞总数与对照组相似,但 B 型精原细胞总数增加59%,而早期(圆形)、中期和晚期(长形)精子细胞总数分别减少37%、72%和52%。结论:EDS 所致精子发生损害主要是(1)精子释放障碍,(2)精子细胞、精母细胞分离并伴有精子形成和成熟分裂障碍。     

白藜芦醇对2,5-己二酮导致SD大鼠睾丸生精障碍治疗作用的实验研究

目的:观察白藜芦醇促进2,5-己二酮(2,5-HD)所致睾丸生精障碍后的生精功能恢复作用。方法:40只雄性SD大鼠随机分为5组(每组8只),A组正常喂饲,B、C、D、E组均饮用含1%2,5-HD的水溶液5周,然后C、D、E组用不同浓度的白藜芦醇[分别为20、40、80 mg/(kg.d)]治疗9周。比较各组外表体征、体重增加值、睾丸重量的变化,及各组生精小管数目、直径以及生精细胞膜c-kit蛋白表达水平。结果:与A组相比,B、C、D、E组大鼠体质瘦弱、皮肤松弛、毛色暗淡、体重增长减慢、睾丸萎缩;睾丸组织HE染色及免疫组化染色显示生精上皮萎缩,生精细胞发育停滞,c-kit蛋白表达受到抑制。经过白藜芦醇治疗后,C、D、E组大鼠外表体征得到恢复,接近A组,体重和睾丸重量均较B组有所恢复(P<0.01);睾丸生精功能部分恢复,但生精小管数目、直径低于A组水平(P<0.001),同时c-kit蛋白重新获得表达,但低于A组水平(P<0.01)。随着白藜芦醇用量增加,C、D、E组大鼠睾丸生精小管数目、直径显著恢复(P<0.01),生精细胞膜c-kit蛋白表达量增加更为明显(P<0.01)。结论:白藜芦醇可以促进2,5-HD所致大鼠睾丸生精障碍的生精功能恢复。     

L-肉碱在奥硝唑所致大鼠睾丸和附睾损伤中的保护作用

目的:探讨L-肉碱(LC)在奥硝唑(ORN)所致大鼠附睾和睾丸损伤中的的保护作用。方法:40只雄性SD大鼠(200～230g)随机均分为5组:①A组:给予0.5%的羧甲基纤维素钠(溶剂)灌胃;②B组:每天给予400mg/kgORN灌胃;③C组:每天给予800mg/kgORN灌胃;④D组:每天给予[ORN(400mg/kg)+LC(100mg/kg)]灌胃;⑤E组:每天给予[ORN(800mg/kg)+LC(100mg/kg)]灌胃。上述各组均连续灌胃20d,末次给药24h后,所有大鼠麻醉后处死,分别取睾丸、附睾,进行称重和HE染色,计算睾丸、附睾系数并观察睾丸和附睾病理组织学改变。结果:①与A组相比,B组睾丸、附睾系数明显降低(P<0.05);而C组睾丸、附睾系数为极显著性降低(P<0.01);D组与A组相比无差异,E组与A组相比有极显著性差异(P<0.01);②HE染色显示,与A组相比,B组睾丸生精小管内各级生精细胞排列基本整齐,部分生精小管管腔内有脱落的生精细胞,附睾管腔中精子数目下降,有时可见散在的生精细胞;C组大鼠睾丸生精小管管腔内均可见坏死脱落的生精细胞,附睾管腔中精子数目明显减少,且有较多的非精子细胞成分。D组睾丸生精小管无明显改变,附睾管腔中精子数目也未见明显下降;E组睾丸生精小管管腔内精子数目减少,可见坏死脱落的生精细胞,附睾腔中精子数目明显减少,并伴有较多的非精子细胞成分。结论:奥硝唑(ORN)可导致雄性大鼠附睾和睾丸病理组织学改变,LC对ORN引起大鼠附睾和睾丸损伤具有一定的保护作用。     

手机频率电磁辐射对大鼠睾丸超微结构损伤的研究

目的:观察手机频率(900 MHz)电磁辐射对雄性大鼠睾丸组织超微结构和生精细胞凋亡的影响。方法:将30只SPF级健康雄性SD大鼠随机分为3组:正常组、辐射2 h组和辐射4 h组,每组10只。正常组不辐射,辐射2 h组和辐射4 h组每天暴露于900 MHz手机频率分别辐射2、4 h,连续辐射30 d。透射电镜观察睾丸组织超微结构的变化,原位末端标记(TUNEL法)检测生精细胞凋亡。结果:与正常组比较,辐射2 h组大鼠生精小管基膜肿胀,支持细胞紧密连接分开,细胞间隙明显增大,部分线粒体内出现空泡,见线粒体髓样化,生精细胞凋亡增多[(25.62±1.33)%vs(18.71±1.76)%,P0.05],以初级精母细胞凋亡为主;与辐射2 h组比较,辐射4 h组生精小管基膜肿胀,支持细胞紧密连接分开,细胞间隙明显增大,精原细胞胞膜不完整,可见胞质碎片、胞核固缩和切迹,核周隙轻度扩张,细胞内线粒体畸形,内有空泡,结构模糊,部分嵴融合或消失,其损伤程度增加,生精细胞凋亡增多[(29.93±1.46)%vs(25.62±1.33)%,P0.05],精原细胞、初级精母细胞和次级精母细胞均有凋亡。结论:手机频率电磁辐射可造成大鼠睾丸组织超微结构损伤,生精细胞凋亡增加,有时效关系;生精细胞凋亡可能与线粒体损伤有关。     

实验性精索静脉曲张大鼠生精状态评价

目的 通过对大鼠实验性精索静脉曲张(varicocele,VC)模型睾丸生精小管生精上皮结构、性激素水平的分析,探讨精索静脉曲张致不育的机制.方法 40只雄性青春期Wistar大鼠随机分为VC8周组(n=12)、VC12周组(13=12)和相应对照组(分别n=8);左肾静脉部分结扎建立实验性大鼠VC模型.术后8周或12周,分别测量各组大鼠:(1)左侧精索静脉直径、睾丸温度及体质量、睾丸生精小管生精上皮;(2)外周血中促卵泡刺激素(FSH)、黄体生成素(LH)和睾酮(T)的水平.结果 VC8周和VC12周组大鼠左侧精索静脉明显扩张,与对照相比血管直径差异有统计学意义(P<0.01);VC8和VC12周组大鼠左侧睾丸体质量均低于自体右侧睾丸和对照组睾丸,差异有统计学意义(P<0.05);光镜下,VC组大鼠双侧睾丸生精小管生精上皮精子发生阻滞、细胞脱落和细胞层数减少等,VC12周组损伤程度较VC8周组明显加重,左侧较右侧显著;与对照组相比,VC组大鼠外周血FSH、LH升高,T降低,差异均有统计学意义(P<0.01).结论 本研究提示:VC对大鼠双侧睾丸生精小管生精上皮产生明显的损害作用,并导致大鼠血中T水平降低和FSH、LH水平升高.     

Attractin蛋白在大鼠睾丸和附睾中的免疫组织化学定位

目的 :探讨Attractin蛋白在成熟大鼠睾丸和附睾组织中的分布。　方法 :健康成年雄性SD大鼠 2 0只 ,灌流取睾丸和附睾组织固定 ,石蜡包埋和冰冻切片。用免疫组化法和间接免疫荧光方法检测Attractin蛋白在成熟大鼠睾丸和附睾组织中的表达。　结果 :睾丸组织间质细胞、睾丸精曲小管管周肌样细胞和各级生精细胞 (精原细胞、初级精母细胞和精子细胞 )、支持细胞呈阳性反应 ,主要表达于胞膜及胞质。睾丸间质细胞表达略强于生精细胞。附睾头、体、尾均未见表达。　结论 :Attractin蛋白在成熟大鼠睾丸组织间质细胞和生精细胞中有较强的表达 ,其生理功能尚有待进一步探讨。     

中药生精冲剂对生精障碍小鼠治疗作用的实验研究

目的:研究生精冲剂对环磷酰胺所致生精障碍小鼠的治疗作用。方法:46只雄性昆明小鼠随机分为正常组(n=10)、模型组(n=12)、对照组(n=12)和中药组(n=12),后3组腹腔注射环磷酰胺,建立生精障碍模型。造模完成后,中药组和对照组分别灌胃中药生精冲剂[16 g/(kg.d)]和克罗米芬[21.6 mg/(kg.d)],正常组与模型组每天灌胃等量生理盐水,连续15 d。次日,测量睾丸重量,并计算睾丸指数;放射免疫法测定血清FSH、LH、T水平;对睾丸组织进行组织学观察。结果:中药组小鼠血清T[(7.046±0.291)nmol/L]、FSH[(2.947±0.587)mIU/ml]、LH[(3.254±0.492)mIU/ml]、睾丸指数[(3.958±0.342)g/kg]与模型组各检测值相比[(6.231±0.317)nmol/L、(5.428±0.719)mIU/ml、(5.155±0.460)mIU/ml、(3.525±0.462)g/kg]差异有显著性(P<0.05)。组织学观察中药组睾丸生精小管生精细胞层数及管腔内精子数明显高于模型组。结论:生精冲剂治疗小鼠生精障碍有效。     

成年大鼠睾丸Smad1和Smad5的免疫组织化学研究

目的 :探讨骨形态形成蛋白的细胞内信号传导分子Smad1和Smad5蛋白在成年大鼠睾丸内的表达。　方法 :应用免疫组织化学ABC法结合葡萄糖氧化酶 DAB 硫酸镍铵增强技术 ,检测成年大鼠睾丸Smad1和Smad5蛋白的表达和定位。　结果 :Smad1蛋白表达于大鼠睾丸精曲小管的各级生精细胞的胞质内 ,呈淡染的紫蓝色颗粒 ,胞核为阴性 ,免疫染色阳性细胞主要包括精原细胞、初级精母细胞、次级精母细胞和精子细胞 ;而Smad5蛋白阳性染色不明显 ,只有睾丸间质细胞呈弱阳性反应 ,免疫反应阳性物质主要定位于细胞质。　结论 :Smad1蛋白主要表达于睾丸精曲小管各级生精细胞 ,Smad5蛋白表达于间质细胞 ,为揭示TGF β超家族在精子发生中的分子机理提供了直接证据。     

酒精染毒后大鼠睾丸生精小管超微结构的改变

目的:观察酒精染毒后大鼠睾丸生精小管超微结构的变化。方法:将48只W istar雄性性成熟健康大鼠随机分为对照组(A组)、染毒组(B组),每组24只,B组用50度酒精(6 m l/kg)每天灌胃染毒1次,连续39 d,A组用生理盐水灌胃(6 m l/kg);在第14、27、40 d分别从A、B两组中各处死8只大鼠,取睾丸组织,常规电镜制片,透射电镜观察大鼠睾丸生精小管超微结构。结果:透射电镜发现:对照组各时间段睾丸生精小管超微结构无明显变化,基膜平滑、致密、厚薄均匀,支持细胞胞质丰富、粗面内织网及线粒体较多,核大、核质均匀、核仁清楚,精原细胞与基膜和支持细胞之间连接紧密,紧密连接层次清楚。染毒组第1生精周期末(相当于第14 d)超微结构开始变化;第2、3个生精周期末(分别相当于第27、40 d)超微结构变化最明显,主要表现为:①支持细胞溶酶体增多,线粒体肿胀空泡化,细胞器模糊且数量减少,支持细胞内出现较大空泡,甚至支持细胞出现萎缩;②精原细胞与支持细胞及生精小管的基膜之间出现较多较大空泡;③精原细胞凋亡,凋亡小体边集;④生精小管内的精子有过多的胞质且有大小不一的空泡,尾部断面线粒体排列不整齐、缺失或堆积;⑤紧密连接层次改变、模糊不清;⑥基膜厚薄不均,基膜外胶原组织疏松增厚,成波浪式皱褶,可见基膜断裂。结论:酒精可引起大鼠睾丸生精小管的基膜、紧密连接、支持细胞等超微结构异常,并可引起精原细胞凋亡。     

五味子对模拟航渡及高强度运动大鼠垂体-睾丸轴及糖代谢的影响

目的:研究五味子对实验性模拟航渡及高强度运动雄性大鼠睾丸轴功能、超微结构及糖代谢的影响。方法:选取34只SD雄性大鼠,分为安静对照组(A组)、应激对照组(B组)、五味子处理组(C组),B组和C组参照Bedford的训练模式进行训练,分别以生理盐水和五味子灌胃1周,每日2次。实验结束后,取血以放免法测定血清睾酮(T)、皮质酮(CORT)、血清黄体生成素(LH)、葡萄糖(G lu)水平,同时随机取垂体、睾丸组织在电子显微镜下观察其超微结构。结果:①A组血G lu为(5.22±2.13)mmol/L,T为(0.61±0.68)ng/m l,CORT为(4.67±1.19)ng/m l;B组血G lu为(9.41±2.56)mmol/L,较A组水平升高(P<0.01),T为(0.10±0.15)ng/m l,与A组相比水平降低(P<0.01),CORT为(7.25±6.20)ng/m l,与A组无明显差异(P>0.05);C组血CORT为(3.55±3.52)ng/m l,G lu为(5.09±1.64)mmol/L,较B组水平均显著降低(P<0.05,P<0.01),T为(0.11±0.12)ng/m l,较B组无明显变化(P>0.05);LH在3组之间均无显著变化(P>0.05)。②电子显微镜观察发现,B组垂体细胞胞质中分泌颗粒较A组显著减少,C组与B组相比胞质中分泌颗粒的数目明显增多,B组睾丸Leyd ig细胞线粒体肿胀,电子密度增高,嵴减少或消失,C组睾丸Leyd ig细胞中线粒体结构趋向于正常,可见大小不等深染的分泌颗粒。结论:模拟航渡及高强度运动训练抑制睾丸功能并影响细胞内超微结构,灌服五味子对垂体-睾丸轴具有保护作用,同时可降低应激大鼠的血糖水平。     

Effects of Oestrogens and FSH on LH Stimulation of Steroid Production by Testis Leydig Cells from Immature Rats

Hypophysectomy of immature male rats results after 5 days in a decreased production of testosterone by isolated testis Leydig cells in response to LH. The LH responsiveness of the Leydig cells can be partly restored by treatment of the hypophysectomized rats with FSH. In continuation of previous reports on this subject ( Steroids 28 (1976) 847; and 30 (1978) the following conclusions were derived from the results in the present paper:
1. After hypophysectomy of immature male rats the production of testosterone (T) as well as of 5-pregnenolone (Δ5P) by isolated Leydig cells in response to LH is reduced.
2. Daily administration of FSH after hypophysectomy restores the Δ5P production in response to LH almost completely, but has a much smaller effect on the restoration of T production.
3. Administration of oestradiol benzoate (E₂B) together with FSH has no effect on the restoration of LH-stimulated Δ5P production, but causes a reduction of T production, when compared with Leydig cells from animals treated with FSH only.
4. Treatment of intact immature rats with E₂B results in a decreased production of T and an increased production of Δ5P in isolated Leydig cells.
5. From experiments with labelled pregnenolone it appears that E₂B and diethylstilboestrol (DES) inhibit the 17α-hydroxylase activity of Leydig cells from intact as well as from hypophysectomized rats. This results in a reduced conversion of pregnenolone to C1:)-steroids and in increased production of 3α-hydroxy-5α-pregnan-20-one from δ5P.
6. The observed effects of FSH and E, were similar within a dose range of 100–10000 ng LH per 106 Leydig cells.     

Expression of leptin and leptin receptor in the testis of fertile and infertile patients

The aim of our study was to investigate the relationships between the expression of leptin, leptin receptor in the testis and spermatogenesis, and testosterone (T) concentration in infertile men. Testicular tissue samples were collected from the testes of five fertile volunteers, eight patients with obstructive azoospermia (OA), six patients with Sertoli cell-only syndrome (SCO) and 32 oligospermic patients with varicocele testis. In testicular tissue, leptin and leptin receptor were identified by staining with polyclonal antibodies. Serum follicle stimulating hormone, lutenising hormone (LH), and T were determined by chemiluminescence assays. Leptin was expressed on germ cells, mainly on spermatocytes. The ratio of immunostained germ cells to total germ cells was inversely correlated with the concentration of T (r = -0.32, P = 0.01), sperm concentration (r = -0.51, P = 0.002) and Johnsen's score (r = -0.44,P = 0.005). In contrast, leptin receptor immunostained cells were found in the interstitium, primarily in Leydig cells. Leptin receptor expression on Leydig cells was inversely correlated with serum T concentration (r = -0.50, P < 0.001). The dysfunction of spermatogenesis is associated with an increase in leptin and leptin receptor expression in the testis.     

Therapeutic effect of osteogenically induced adipose derived stem cells on vascular deprivation-induced osteonecrosis of the femoral head in rabbits

Objective: To explore the therapeutic effect of osteogenically induced adipose-derived stem cells (ADSCs) on vascular deprivation-induced osteonecrosis of the femoral head (ONFH) in rabbit model. Methods: Vascular deprivation-induced ONFH was established by intramuscular injection of methylprednisolone, and vascular occlusion of the capital femoral epiphysis by electrocoagulation in adult New Zealand white rabbits. Eight weeks after the establishment of vascular deprivation-induced ONFH, animals were randomly divided into three equal groups. In Group A (control), no therapy was given. In Group B, core decompression was performed by drilling a hole (1.2 mm in diameter) from the outer cortex 2.5 cm distal to the proximal end of the greater trochanter. In Group C, 1 ×107 osteogenically induced ADSCs were resuspended in 0.5 ml PBS, and then injected directly into the femoral head. Femoral head specimens were obtained at postoperative 8 weeks. The bone formation and three-dimensional microstructure of the femoral head was evaluated by micro-computed tomography scans (μ-CT). Immunohistochemical analysis was performed to detect the expression of osteocalcin. Angiogenesis and repair of the femoral head were observed histologically. Results: In trabecular bone at the proximal femur region, the trabecular volume was higher in Group C (130.70 mm3± 4.33 mm3) than that in Groups A (101.07 mm3±7.76 mm3) and B (107.89 mm3±8.68 mm3, P＜0.01). Bone volume was significantly increased in Group C (40.09 mm3±6.35 mm3) than in Groups A (29.65 mm3±4.61 mm3) and B (31.80 mm3± 4.01 mm3, P＜0.01). The trabecular number was higher in Groups C (1.58±0.25) than other two groups (1.15±0.18, 1.16± 0.21, P＜0.01). Bone mineral density showed statistically significant difference between Groups C and A or B (375.38± 23.06) mg HA/ccm, vs (313.73 ±19.30) mg HA/ccm and (316.09± 16.45) mg HA/ccm, P<0.01). Histological examination indicated that there was more new bone formation in Group C than in other groups. Conclusion: Treatment with autologous osteogenically induced ADSCs transplantation results in an enhanced osteogenesis and microstructure of the vascular deprivation-induced osteonecrosis in rabbits.     

藏药鲁堆多吉水提物对雄性幼年大鼠生殖系统的影响

目的:研究藏药鲁堆多吉对雄性幼年大鼠生殖系统的影响。方法:将32只SD雄性幼年大鼠分为对照组、鲁堆多吉水提物低、中、高剂量组,灌胃2周后处死,用放射免疫法检测大鼠血清中睾酮(T)的含量,用ELISA试剂盒测量大鼠血清中LH和FSH的含量,称量大鼠的睾丸重量,计算体重增加量、睾丸脏器系数和精子浓度。将高剂量组大鼠血清在57℃水浴锅中灭活后,按0(对照组)、10%、15%、20%的体积分数加到培养的大鼠睾丸间质细胞中,分别用台盼蓝染色法和MTT检测细胞活率和活力,用放免法检测睾丸间质细胞培养液中T的含量,用大鼠环磷酸腺苷(cAMP)ELISA试剂盒检测睾丸间质细胞内cAMP的变化。结果:与对照组相比,低、中、高剂量的鲁堆多吉水提物能使幼年雄性大鼠的血清T水平[(1.22±0.32)、(1.06±0.29)、(0.57±0.18)nmol/L vs(3.09±0.42)nmol/L,P0.01]、睾丸重量[(0.96±0.09)、(0.92±0.11)、(0.91±0.08)g vs(1.40±0.16)g,P0.01]和精子浓度[(0.19±0.07)、(0.17±0.08)、(0.16±0.07)×106/ml vs(1.03±0.16)×106/ml,P0.01]显著降低,并且呈剂量依赖性。与对照组相比,中、高剂量给药组血清中的LH[(14.69±0.12)、(14.93±0.28)ng/L vs(13.62±0.89)ng/L,P0.01]、FSH[(4.77±0.23)、(4.89±0.38)IU/L vs(4.32±0.18)IU/L,P0.05]显著上升;睾丸脏器系数[(51.39±3.09)、(52.28±4.86)、(54.13±6.06)mg/10 g vs(42.22±3.02)mg/10 g,P0.01]显著提高。与对照组相比,鲁堆多吉水提物含药血清能显著抑制睾丸间质细胞内cAMP水平[(4.39±0.06)、(4.28±0.07)、(4.11±0.10)nmol/L vs(5.51±0.12)nmol/L,P0.01]和T的合成[(1.42±0.15)、(1.12±0.18)、(0.88±0.21)nmol/L vs(1.85±0.18)nmol/L,P0.01]。结论:鲁堆多吉水提物可能通过PKA途径抑制大鼠睾丸间质细胞中T的合成,从而抑制睾丸发育和精子生成,影响雄性幼鼠生殖系统的发育。     

Effects of supplementation with impuberal or adult testicular protein extracts on genital tract and testicular histology as well as hormonal levels in adult busulfan treated rats

24 adult Wistar rats received one SC injection of busulfan (10 mg/kg BW) on day 0. They were injected daily from day 1 to day 10 either with BSA (1 and 2 mg/rat, n = 5 respectively), with acetonic extract of adult ram testicular cytosol (2 mg/rat, n = 10) or with acetonic extract of impuberal calf testicular cytosol (2 mg/rat, n = 4). 10 animals served as control. Animals were killed on day 17. Busulfan treatment induced decrease of total volumes of intertubular tissue and of Leydig cells per testis, of accessory glands, and of germ cells from type A spermatogonia to zygotene spermatocytes. Supplementation with adult testicular extract did not modify accessory glands but increased Leydig cell total volume, individual cellular and nuclear areas and Sertoli cell nuclear areas. It decreased further the germ cells from A spermatogonia to zygotene primary spermatocytes. Supplementation with impuberal testicular extract did not modify accessory glands, Leydig cell total volume, individual cellular and nuclear areas or Sertoli cell nuclear area. It increased the germ cells from A spermatogonia to zygotene primary spermatocytes to values intermediate between those of control and busulfan treated rats.     

实验性水上漂浮和高强度运动对大鼠下丘脑-垂体-睾丸轴功能的影响

目的:探讨大鼠在经历水上漂浮和高强度运动后血清T、皮质酮(CORT)、LH、FSH的变化规律。方法:将大鼠随机分为对照组(C组)、水上漂浮组(A组)、水上漂浮并高强度运动组(B组)3组,每组12只,实验分为3个阶段:第一阶段为适应性训练阶段,3d,每天10min,对模拟水上漂浮装置和大鼠跑台进行适应性训练;第二阶段为水上漂浮阶段,共3h;第三阶段为水上漂浮并高强度运动阶段,共2h。A组大鼠和B组大鼠分别经历第一、二阶段和第一、二、三阶段。实验结束后测定血清T、CORT、LH、FSH水平。结果:血清T(呈非正态分布,经对数转换后分析)A组为0.17±0.07,B组-1.11±0.35,与C组0.67±0.26相比均显著下降(P<0.05);LHB组为(3.97±1.57)mIU/ml,与C组(6.49±1.56)mIU/ml相比显著下降(P<0.05);FSH无显著变化;CORTB组为(977.22±207.36)ng/ml,与C组(434.58±110.45)ng/ml相比明显上升(P<0.05)。结论:在水上漂浮阶段,大鼠血清T出现明显下降,说明严重的心理应激会抑制下丘脑-垂体-睾丸轴的功能。随着应激原的增多和应激强度的增加,LH明显下降,T进行性下降,说明下丘脑-垂体-睾丸轴功能受到进一步抑制。     

Hormonal regulation of inhibin B secretion by immature rat sertoli cells in vitro: possible use as a bioassay for estrogen detection

The influences of follicle-stimulating hormone (FSH), gonadal steroids, and culture time were studied in relation to inhibin B production by Sertoli cells of immature rats cultured in vitro. Sertoli cell-enriched cultures were established from 18-day-old rats and were maintained in medium supplemented with insulin, transferrin, and epidermal growth factor at 34 degrees C. A recently developed ELISA for the measurement of inhibin B was used to assess the effects of recombinant human FSH (rh FSH), testosterone (T), and estradiol (E2) on inhibin B production and accumulation in the culture media of Sertoli cell-enriched cultures and to optimize the cell culture system to serve as a bioassay for the detection and quantification of estrogens and estrogenlike substances. Prolonging the incubation time (24, 48, or 72 hours) of Sertoli cells with control medium without rh FSH, T, or E2 resulted in a time-dependent increase of inhibin B production. Incubation with rh FSH (1, 2.5, 5, or 10 U/L) caused a dose- and time-dependent increase of inhibin B production by Sertoli cells (but not by cultured Leydig cells), reaching a plateau at 5 U/L rh FSH. Addition of T in concentrations of 2.88, 5, or 50 ng/ml to medium without rh FSH and E2 significantly lowered the daily production rate of inhibin B (P < 0.05). In contrast, addition of E2 (0.01 and 0.1 ng/ml) caused a dose-responsive increase in inhibin B production after 24 and 48 hours. The relative increment of inhibin B production induced by E2 was maximal after 24 hours in the presence of 2.5 U/L rh FSH (acting synergistically) and in the absence of T. When these conditions are implemented, the Sertoli cell culture system may serve as a bioassay for estrogenic substances, and it may reflect the possibly harmful effect they may have on spermatogenesis.     

Temperature and germ cell regulation of Leydig cell proliferation stimulated by Sertoli cell-secreted mitogenic factor: a possible role in cryptorchidism

Summary. Local control of Leydig cell morphology and function by seminiferous tubules was suggested in previous in vivo studies, especially those that used experimental cryptorchid rat testis as a model. These studies reported changes in morphology, increases in cell number and mitotic index and decreases in testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels of Leydig cells. However, little is known about how these changes are mediated. We recently observed that a novel Sertoli cell-secreted mitogenic factor stimulated proliferation, decreased testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels, and dramatically altered the morphology of Leydig cells in culture. In the present studies, we demonstrate that an increase in coculture temperature from 33 to 37 °C increased [³H]-thymidine incorporation (5.6- vs. 19.2-fold) and labelling index (4.3% vs. 15.8%), and accelerated proliferation (2.1- vs. 3.9-fold) of cultured immature Leydig cells. In addition, testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels of Leydig cells cocultured with Sertoli cells were further decreased following a 4°C increase in coculture temperature. This elevation in culture temperature increased both the secretion of this factor by Sertoli cells and responsiveness of Leydig cells to this factor. In addition, the presence of germ cells, especially pachytene spermatocytes, inhibited the secretion of the mitogenic factor by Sertoli cells. These temperature- and germ cell-associated effects mimicked the morphological and functional changes of Leydig cells reported following experimental cryptorchidism. These observations suggest a possible role of this Sertoli cell-secreted mitogenic factor in explaining Leydig cell changes following experimental cryptorchidism.     

Reproductive effects of the anticancer drug cyclophosphamide in male rats at different ages

This study was undertaken to determine the effects of the anticancer and immunosuppressant drug cyclophosphamide (CP) on several endpoints of the male rat reproductive system at different ages; 10-day-old (experiment A), 45-day-old (experiment B), and adult (experiment C) Sprague-Dawley rats were injected intraperitoneally with CP at doses of 20 mg/kg/day or/week and 100 mg/kg/week for 2 weeks (experiment A), doses of 20 mg/kg/day for 5 weeks and 100 mg/kg/day for 10 days (experiment B), and doses of 20 mg/kg/day for 5 weeks (experiment C). In all groups CP induced a significant rate of mortality. Body weight gain was moderately to severely reduced in two groups of experiment A (20 mg/kg/day and 100 mg/kg/week) and of experiment B (20 mg and 100 mg/kg/day) but normal in the others. Absolute as well as relative reproductive organ weights decreased following some of the treatments in experiments A and B. At the light microscope level, effects of CP ranged from nonapparent in immature rats (experiment A, 100 mg/kg/week for 2 weeks) and young adult animals (experiment B, 100 mg/kg/day for 10 days) to moderate in the other groups treated for 5 weeks (experiments B and C). Affected tubules exhibited atrophy, exfoliation, and a decrease in the number of spermatogonia, primary spermatocytes, and round and elongated spermatids. Sertoli cell function appeared preserved, whereas Leydig cells, present in the intratubular tissue of the rats in all the experiments, were occasionally and moderately altered in animals of experiment B, as shown by significant decreases of serum testosterone and LH levels. Leydig cell dysfunction in these rats was associated with normal in vitro basal and hCG-stimulated testosterone production. A significant decrease in epididymal sperm reserves was observed only in one group of animals (experiment B, 100 mg/kg/day for 10 days). Since in these animals the number of spermatids in the seminiferous tubules was normal, it is possible that CP at a high dose alters the epididymal function. Furthermore, fertility trials demonstrated that despite no change in the number of implantation sites, there was a dramatic fall in the number of fetuses per female in all the experimental groups. In conclusion, this study shows that in pre- and postpubertal rats treated chronically or subacutely, CP primarily and essentially induces alterations of germ cells, whereas this compound has little or no direct effect upon Leydig cell and Sertoli cell functions, respectively.