Horsfall D J, Mayne K, Skinner J M, Saccone G T, Marshall V R, Tilley W D
Department of Surgery, Flinders University School of Medicine, Flinders Medical Centre, Adelaide, Australia.
Prostate. 1994 Dec;25(6):320-32. doi: 10.1002/pros.2990250606.
The effects of aging and hormone manipulation on the glycosaminoglycan (GAG) content of prostatic stroma in guinea pigs were investigated. Total GAG and individual GAG classes (chondroitin, dermatan, and heparan sulfates, and hyaluronic acid) were measured biochemically in stromal extracts. Chondroitin sulfate was also measured and localized by video image analysis of immunocytochemically-stained tissue sections. The weight and total GAG (uronic acid) content of prostatic stroma increased between the ages of 2 weeks and 2 years by 7-8-fold and 4-5-fold respectively. GAG concentration per unit weight of stroma declined 4-fold during puberty and remained essentially unchanged thereafter. Similar results were obtained for each of the GAG classes. The decreases in GAG concentration were associated with a 3-fold increase in the size of the smooth muscle cells of the prostatic stroma during puberty. Hormonal control of GAG deposition in the prostatic stroma was investigated by steroid replacement in prepubertally-castrated animals. Administration of dihydrotestosterone (DHT) to castrate animals for 6 weeks resulted in significantly reduced concentrations of stromal uronic acid, compared with untreated castrate animals (P < 0.05). The GAG levels post-DHT treatment were similar to those observed after pubertal development in sham-operated control animals. Estradiol treatment had the opposite effect to that of DHT, resulting in a significantly increased concentration of uronic acid compared with castrate animals (P < 0.05). These steroid-induced changes in stromal GAG deposition were mostly contributed to by chondroitin and dermatan sulfates. Combined treatment with DHT and estradiol resulted in stromal uronic acid concentrations similar to those of animals receiving DHT alone, indicating that the effect of DHT on stromal GAG deposition is dominant over the effects of estradiol. Morphometric measurement, using computer-assisted video image analysis of a chondroitin sulfate epitope in prostatic sections stained with a monoclonal antibody (6C3), supported the biochemical data. Stereometric profiles across several sectioned glands demonstrated that chondroitin sulfate was confined to the periacinar basement membranes of the prostatic stroma in all groups except the estradiol-treated castrate animals, where the immunostaining extended from the periacinar basement membrane throughout the fibromuscular stroma. Treatment of castrate animals with estradiol alone also induced a physicochemical change in the chondroitin sulfate molecule, resulting in reduced electrophoretic mobility. In summary, this study identifies changes in the quantity, structure, and localization of chondroitin sulfate in the prostatic stroma of estradiol-treated guinea pigs. Furthermore, estradiol and DHT have opposing effects on the level of chondroitin and dermatan sulfate expression in the prostatic stroma.
研究了衰老和激素调控对豚鼠前列腺基质中糖胺聚糖（GAG）含量的影响。采用生化方法测定了基质提取物中的总GAG以及各类型GAG（硫酸软骨素、硫酸皮肤素、硫酸乙酰肝素和透明质酸）。还通过对免疫细胞化学染色组织切片的视频图像分析来测定和定位硫酸软骨素。前列腺基质的重量和总GAG（糖醛酸）含量在2周龄至2岁之间分别增加了7 - 8倍和4 - 5倍。每单位重量基质中的GAG浓度在青春期下降了4倍，此后基本保持不变。各类型GAG均得到类似结果。GAG浓度的降低与青春期前列腺基质平滑肌细胞大小增加3倍有关。通过对青春期前阉割动物进行类固醇替代来研究激素对前列腺基质中GAG沉积的调控。给阉割动物注射二氢睾酮（DHT）6周后，与未处理的阉割动物相比，基质糖醛酸浓度显著降低（P < 0.05）。DHT处理后的GAG水平与假手术对照动物青春期发育后的水平相似。雌二醇处理产生与DHT相反的效果，与阉割动物相比，糖醛酸浓度显著增加（P < 0.05）。这些类固醇诱导的基质GAG沉积变化主要由硫酸软骨素和硫酸皮肤素引起。DHT和雌二醇联合处理导致基质糖醛酸浓度与单独接受DHT的动物相似，表明DHT对基质GAG沉积的影响比雌二醇的影响更显著。使用计算机辅助视频图像分析对用单克隆抗体（6C3）染色的前列腺切片中的硫酸软骨素表位进行形态测量，支持了生化数据。对多个切片腺体的立体测量轮廓显示，除了雌二醇处理的阉割动物外，所有组中硫酸软骨素都局限于前列腺基质的腺泡周围基底膜，在雌二醇处理的阉割动物中，免疫染色从腺泡周围基底膜延伸至整个纤维肌性基质。单独用雌二醇处理阉割动物也诱导了硫酸软骨素分子的理化变化，导致电泳迁移率降低。总之，本研究确定了雌二醇处理的豚鼠前列腺基质中硫酸软骨素在数量、结构和定位上的变化。此外，雌二醇和DHT对前列腺基质中硫酸软骨素和硫酸皮肤素的表达水平具有相反的作用。
