摘要
目的:研究Wnt/β-catenin信号通路在去睾丸小鼠骨质疏松发病中的作用。方法:11月龄雄性健康ICR小鼠30只,随机分为3组,睾丸切除组、睾丸切除+雄激素替代组和假手术组,每组10只。睾丸切除组和睾丸切除+雄激素替代组摘除小鼠双侧睾丸,假手术组仅切开皮肤,分离睾丸包膜。手术后睾丸切除+雄激素替代组丙酸睾酮肌注每次10μg,每隔3 d注射1次;睾丸切除组和假手术组0.9%氯化钠溶液肌注每次10μg,每隔3 d注射1次。饲养14周后,检测3组小鼠骨密度(BMD)。处死后心脏采血,ELISA法检测雌、雄激素含量。取小鼠骨组织,部分提取RNA并翻转成为c DNA,再通过实时荧光定量PCR法检测β-catenin和Runx2的m RNA表达;部分提取总蛋白,通过Western blot法检测β-catenin和Runx2蛋白的表达。结果:与假手术组比,睾丸切除+雄激素替代组和睾丸切除组BMD值均降低,差异均有统计学意义(P<0.05);睾丸切除+雄激素替代组与睾丸切除组比较BMD值升高,差异有统计学意义(P<0.05)。与假手术组比,睾丸切除组小鼠β-catenin和Runx2的m RNA和蛋白表达均降低,差异有统计学意义(P<0.05);使用雄激素替代治疗后,β-catenin和Runx2的m RNA及蛋白表达均上调,差异有统计学意义(P<0.05)。结论:去睾丸骨质疏松小鼠Wnt/β-catenin信号通路受到抑制,可能是其骨质疏松发生发展的重要分子生物学机制之一。
Objective： To evaluate the potential pole of Wnt/beta-catenin signaling pathway in the induc-tion and development of osteoporosis within elder mice. Methods： Healthy ICR mice were randomly divided into three groups, including sham-operated group （n=10）, orchiectomy （ORX） group （n=10） and ORX＋androgen group （n=10）. Testicles were removed from the mice within ORX group and ORX＋androgen group. In sham-operated group, the surgery to separate the testicular capsula was performed and the testicles were intact. After surgery, all the mice were fed for 14 weeks freely with water and food under the same condition. The mice were subjected to the evaluation of the bone mineral density （BMD）. The blood samples were collected from heart to examine the expression level of estrogen and androgen by an ELISA method. The bone tissues were obtained. The expression levels of β-catenin and Runx2 mRNA expression were determined by real-time PCR while the expression levels of β-catenin and Runx2 protein were evaluated by Western blot. Results： The BMD in the mice of ORX group was significantly decreased compared with NS group （P〈0.05）. The expression levels of bothβ-catenin and Runx2 in the mice of ORX group were also significantly decreased compared with sham-operated group （P〈0.05）. The mice in ORX＋androgen group showed no significant difference from ORX group （P〉0.05） in BMD. The treatment of androgen could enhance the expression levels of β-catenin and Runx2 in ORX group. Conclusion： Wnt/β-catenin signaling pathway is inhibited 14 weeks after orchiectomy in osteoporosis mice, which can be involved in the induction and development of the osteoporosis.
出处
《温州医科大学学报》
CAS
2017年第7期519-522,共4页
Journal of Wenzhou Medical University
