摘要
目的建立成骨细胞体外培养模型,并研究雌二醇(E_2)对体外培养大鼠成骨细胞主要凋亡受体mRNA的调节作用,探讨E_2抑制成骨细胞凋亡的机制。方法用新生大鼠颅盖骨进行成骨细胞的原代培养并进行纯化和鉴定;应用TNF-α(200U/ml)以及TNF-α+E_2(10^(-8)mol/L)进行刺激;用AO-EB染色观察成骨细胞凋亡并计数和计算凋亡率;应用RT-PCR的方法测定成骨细胞的Fas、FasL、TNFR1、TNFR2 mRNA的相对表达量。结果 10^(-8)mol/L的E_2明显抑制由TNF-α诱导的成骨细胞Fas、TNFR1 mRNA的表达(P<0.01),但是对FasL、TNFR2 mRNA的表达没有影响(P>0.05)。结论雌激素可以通过调节成骨细胞凋亡受体的表达来改变成骨细胞对致死性细胞因子如TNF-α和FasL等的凋亡敏感性,从而抑制其凋亡。选择性抑制成骨细胞凋亡受体的表达可能有助于增加成骨。
Objective To evaluate the effects of E2 on the expressions of major Death Receptors (DRs) mRNA in osteoblastes (OBs) , and to explore the mechanism of anti-apoptosis effects of E2 on OBs. Methods OBs from neonatal rats' cranium were cultured and then purified and appraised. They were treated with TNF-α (200U/ml) or TNF-α + E2 (10-gmol/L) for 24h. The apoptotie cells stained by AO-EB were counted. The expressions of mRNAs for Fas, FasL, TNFR1 and TNFR2 in OBs were assayed by RT-PCR. Results E2 ( 10^-8mol/L) suppressed the expressions of Fas and TNFR1 mRNAs in OBs induced by TNF-α (200U/ml) (P 〈0. 01 ), and had no inhibitive effects on the expressions of FasL and TNFR2 mRNAs in OBs indueed by TNF-a (200U/ml) ( P 〉 0. 05 ). Conclusion E2 could downregulate the apoptosis sensitivity of OBs to TNF-α and FasL. Selectively inhibiting the expressions of DRs of OBs may contribute to the treatment of osteoporosis.
出处
《中华骨质疏松和骨矿盐疾病杂志》
2009年第3期183-188,共6页
Chinese Journal Of Osteoporosis And Bone Mineral Research
关键词
雌激素
成骨细胞
凋亡
骨质疏松
estradiol ( E2 )
osteoblaste
apoptosis
osteporosis