Estrogens attenuate oxidative stress and the differentiation and apoptosis of osteoblasts by DNA-binding-independent actions of the ERalpha.
Estrogens diminish oxidative stress in bone and bone marrow, attenuate the generation of osteoblasts, and decrease the prevalence of mature osteoblast apoptosis. We have searched for the molecular mechanism of these effects using as tools a mouse model bearing an estrogen receptor alpha (ERalpha) knock-in mutation that prevents binding to DNA (ERalpha(NERKI/-)) and several osteoblast progenitor cell models expressing the wild-type ERalpha or the ERalpha(NERKI/-). We report that the ability of estrogens to diminish the generation of reactive oxygen species, stimulate the activity of glutathione reductase, and decrease the phosphorylation of p66(shc), as well as osteoblastogenesis and osteoblast number and apoptosis, were fully preserved in ERalpha(NERKI/-) mice, indicating that the DNA-binding function of the ERalpha is dispensable for all these effects. Consistent with the attenuation of osteoblastogenesis in this animal model, 17beta-estradiol attenuated bone morphogenetic protein 2 (BMP-2)-induced gene transcription and osteoblast commitment and differentiation in murine and human osteoblastic cell lines. Moreover, 17beta-estradiol attenuated BMP-2-induced differentiation of primary cultures of calvaria- or bone marrow-derived osteoblastic cells from ERalpha(NERKI/-) mice as effectively as in cells from wild-type littermates. The inhibitory effect of the hormone on BMP-2 signaling resulted from an ERalpha-mediated activation of ERKs and the phosphorylation of Smad1 at the linker region of the protein, which leads to proteasomal degradation. These results illustrate that the effects of estrogens on oxidative stress and the birth and death of osteoblasts do not require the binding of ERalpha to DNA response elements, but instead they result from the activation of cytoplasmic kinases.
Almeida
Maria
M
Division of Endocrinology and Metabolism, Center for Osteoporosis and Metabolic Bone Diseases, University of Arkansas for Medical Sciences and Central Arkansas Veterans Health Care System, Little Rock, AR, USA.
Martin-Millan
Marta
M
Ambrogini
Elena
E
Bradsher
Robert
R
Han
Li
L
Chen
Xiao-Dong
XD
Roberson
Paula K
PK
Weinstein
Robert S
RS
O'Brien
Charles A
CA
Jilka
Robert L
RL
Manolagas
Stavros C
SC
D000818:Animals; D017209:Apoptosis; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation; D004789:Enzyme Activation; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000378:metabolism* / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000032:analysis; D005260:Female; D055879:Gene Knock-In Techniques; D005980:Glutathione Reductase / Q000032:analysis; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D010012:Osteogenesis / Q000187:drug effects; D018384:Oxidative Stress; D010766:Phosphorylation; D055765:Shc Signaling Adaptor Proteins / Q000378:metabolism; D051898:Smad1 Protein / Q000032:analysis; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D014158:Transcription, Genetic / Q000187:drug effects
C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D047628:Estrogen Receptor alpha; D004967:Estrogens; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D051898:Smad1 Protein; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D005980:Glutathione Reductase; D048049:Extracellular Signal-Regulated MAP Kinases