The effects of sex hormone on growth plate chondrocyte.

性激素对生长板软骨细胞的影响。

• 批准号: 14571355
• 负责人: YAMADA Norikazu
• 金额: $ 2.18万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571355/
• 关键词: Growth Plate chondrocyte; Castration; Testosterone; Apoptosis; エストロゲン

Sex hormones play important roles in the regulation of chondrocyte proliferation, maturation and death in epiphyseal growth plate. We investigated the effects of male castration on the cell kinetics of chondrocytes, defined by the numbers of proliferating and dying cells. The growth plates of normal rabbits and castrated at 8 weeks of age were obtained at 10, 15, 20 and 25 weeks old. They were stained for apoptotic and proliferating chondrocytes using immunostaining for caspase-3 and PCNA. The castrated rabbits showed higher caspase-3-positive and lower PCNA-positive ratios than the corresponding normal groups. In addition, the number of chondrocytes in the castrated rabbits was less than that of the normal ones at the same age. Our study suggested that castration led an increase of apoptosis and decrease of proliferating ability in growth plate chondrocytes.Then, we cultured chondrocytes to investigate the effects of sex hormone on chondrocytes in vitro. 17-day-old fatal chick sterno was obtained and revealed by H-E stain to be composed of hypertrophic chondrocytes in 1/4 of cranial side and premature chondrocytes in 1/2 of caudal side. We obtained 5mm cranial and 8mm caudal side of sterno and cultivate them according to Gerstenfeld's method. The RNA of the cultured chondrocytes was extracted and reversetranscription-polymerase chain reaction (RT-PCR) was performed. The cranial chondrocytes presented type 10 collagen and the caudal ones type 2 collagen. That indicated that the former is mature hypertrophic chondrocyte and the latter premature chondrocyte. Sex hormone load test was performed with the culture system. Testosterone and estrogen at 10-10,10-9,10-8,10-7,10-6mol/l were added in hypertrophic and premature chondrocyte. The ALP activity, proliferative activity, cell death, mRNA of type 2 and 10 collage were tested. Testosterone stimulated cell proliferation, and estrogen stimulated cell death.

性激素在骨骺生长板软骨细胞增殖、成熟和死亡的调控中起重要作用。我们研究了雄性阉割对软骨细胞动力学的影响，通过增殖和死亡细胞的数量来定义。分别于10、15、20、25周龄取正常兔和8周龄去势兔的生长板。用caspase-3和PCNA免疫染色法检测细胞凋亡和增殖软骨细胞。与正常对照组相比，去势组caspase-3阳性比例较高，pcna阳性比例较低。此外，去势家兔的软骨细胞数量比同龄正常家兔少。本研究提示去势导致生长板软骨细胞凋亡增加，增殖能力下降。然后，体外培养软骨细胞，研究性激素对软骨细胞的影响。H-E染色显示，17日龄死亡鸡胸骨由1/4的颅侧肥大软骨细胞和1/2的尾侧过早软骨细胞组成。取胸骨头侧5mm、尾侧8mm，按Gerstenfeld法培养。提取培养的软骨细胞RNA，进行逆转录聚合酶链反应（RT-PCR）。颅骨软骨细胞呈10型胶原，尾骨软骨细胞呈2型胶原。提示前者为成熟肥大软骨细胞，后者为早熟软骨细胞。用培养系统进行性激素负荷试验。在肥大和过早软骨细胞中分别添加10-10、10-9、10-8、10-7、10-6mol/l的睾酮和雌激素。检测2型和10型细胞的ALP活性、增殖活性、细胞死亡、mRNA水平。睾酮刺激细胞增殖，雌激素刺激细胞死亡。