摘要
目的观察17β-雌二醇和孕酮对人成骨细胞和人成骨肉瘤MG-63细胞胰岛素受体底物家族 (insulin receptor substrate IRS)表达的影响。探讨雌、孕激素对骨的作用机制。方法从正常骨折病人骨组织中分离正常人成骨细胞,经纯化和培养后,用Van Gieson行Ⅰ型胶原染色、钙钴法行碱性磷酸酶染色、茜素红行钙化结节染色进行成骨细胞鉴定。用半定量RT-PCR检测细胞IRS-1、- 2、-3 mRNA表达量。结果分离和培养的细胞具有正常人成骨细胞的形态,能分泌Ⅰ型胶原、碱性磷酸酶并形成钙化结节。17β-雌二醇和／或孕酮均不影响人成骨细胞和MG-63细胞IRS-1mRNA 的表达(P>0．05),可诱导人成骨细胞和MG-63细胞IRS-2 mRNA的表达上调(P<0．05),IRS- 3 mRNA的表达下调(P<0．05)。二者联合干预时作用明显加强(P<0．01)。结论 17β-雌二醇和孕酮均不影响IRS-1 mRNA的表达,但可使人成骨细胞和MC-63细胞IRS-2 mRNA的表达上调, IRS-3mRNA的表达下调,二者联合干预具有正性协同效应。不同的IRS家族成员对同一细胞具有效应的多样性。
Objective To investigate the effects of 17β-estradiol (E2) and progesterone (P) on the expression gene of insulin receptor substrate (IRS) family in cultured normal human osteoblast-like cells (HOB) and human osteosarcoma cell line MG-63 and its mechanism. Methods Human osteoblasts were isolated from bone tissues in subjects free of osteoporotic fractures. Alkaline phosphatase, mineralized nodes, and type 1 collagen in osteoblasts were tested by modified staining techniques of Gomori' s, alizarin red, and Van Gieson, respectively. Semi-quantitative RT-PCR was used to measure on the expression of E2 and P mRNA of IRS family. Results No influence was found of E2 and P on IRS-1 mRNA expression in MG-63 cells or HOB. The expression of IRS-2 mRNA ( P<0. 05) was increased and the expression of IRS-3 mRNA was decreased by E2 and/or P (P<0. 05) . This effect became mone significant with the combination of E2 and P. Conclusions E2 and P have no effects on the expression of IRS-1 mRNA in MG-63 cells and HOB, but could up-regulate the expression of IRS-2 mRNA and down -regulate the expression of IRS-3 mRNA. Combined intervention with E2 and P can positively create synergistic effects on expressions of IRS-2 and IRS-3.
出处
《中国骨质疏松杂志》
CAS
CSCD
2005年第2期140-145,共6页
Chinese Journal of Osteoporosis
基金
国家自然科学基金资助项目(30400218)
