摘要
目的观察雌激素对绝经后骨质疏松患者骨髓间充质干细胞(OP-hBMSCs)Notch信号通路的影响。方法无菌条件下抽取绝经后骨质疏松患者和健康女性骨髓间充质干细胞(hBMSCs),分离培养并传代。通过流式细胞仪分子表型鉴定确定培养的细胞为骨髓间充质干细胞。比较绝经后骨质疏松患者与健康女性骨髓间充质干细胞的生物特征。通过实时定量-聚合酶链式反应(RT-PCR)比较OP-hBMSCs与hBMSCs中Notch信号通路关键分子(Notch1、Jagged1、Hes1)的表达水平。对人骨髓间充质干细胞进行成骨诱导与成脂诱导,观察雌激素对Notch信号通路Notch1,Jagged1、Hes1表达的影响。结果与正常健康女性相比,绝经后骨质疏松患者骨髓间充质干细胞Notch信号通路减弱(P<0.01)。给予雌激素后,可逆转绝经后骨质疏松患者骨髓间充质干细胞已减弱的Notch信号通路活性,使Notch信号通路下游分子Hes1表达上升近3倍。雌激素促进OP-hBMSCs向成骨分化,抑制成脂分化,Notch信号通路关键分子的表达均明显增高(P<0.01)。结论 Notch信号通路在绝经后骨质疏松患者骨髓间充质干细胞显著减弱。Notch信号通路可能在绝经后骨质疏松症的发生过程中起着重要作用,可能成为雌激素治疗绝经后骨质疏松症的新机制。
Objective To study the effects of estrogen on the activity of Notch signaling in hBMSCs from patients with postmenopausal osteoporosis. Methods hBMSCs were obtained from healthy women and patients with postmenopausal osteoporosis after informed consent to the research protocol. We characterized several phenotypes known to be associated with hBMSCs by flow cytometric analysis of expressed surface markers. Real-time PCR was performed to determine the expression level of key molecules of Notch signaling in hBMSCs between patients with postmenopausal osteoporosis and healthy women. We further tested the relationship between estrogen and Notch signaling during the osteoblastic dif- ferentiation and adipogenic differentiation of hBMSCs. Real-time PCR was performed to evaluate the expression of Notchl, Jaggedl and Hesl. Results The key molecules of Notch signaling impaired in hBMSCs from postmenopausal osteoporosis compared with those from healthy women （P 〈 0. 01 ）. When OP-hBMSCs were treated with 17β-estradiol, the expression of Notchl and Jagged1 were found to be significantly up-regulated. Hesl, which is known as a target of canonical Notch signaling, also demonstrated a more than 3-fold increase in its expression level. Real-time PCR was performed to demonstrate that 17β-estradiol enhanced the expression of the expression level of key Notch signaling molecules during osteoblastic differentiation and adipogenic differentiation of hBMSCs. Meanwhile, estrogen can promote osteogenesis and inhibit adipogenesis. Conclusion The results showed that the key molecules of Notch signaling impaired significantly in hBMSCs from postmenopausal osteoporosis. The present study provides the foundation for further study on the effect of Notch signaling in pathogenesis of postmenopausal osteoporosis.
出处
《中华骨质疏松和骨矿盐疾病杂志》
2013年第3期232-239,共8页
Chinese Journal Of Osteoporosis And Bone Mineral Research
基金
国家重点基础研究发展计划(2011CB9647O3-G)
