Adipocyte differentiation in Sod2(-/-) and Sod2(+/+) murine bone marrow stromal cells is associated with low antioxidant pools.
OBJECTIVE
Adipocytogenesis in bone marrow stromal cells (BMSCs) from manganese-superoxide dismutase-deficient (Sod2(-/-)) and wild-type (Sod2(+/+)) mice and the effect of antioxidant pool size were determined.


METHODS
BMSCs from Sod2(-/-) or Sod2(+/+) mice were cultured with and without adipocytogenic supplements including: 10 mug/mL insulin, 1 muM dexamethasone, and 100 muM indomethacin. Oil Red-O-positive cells and reverse-transcriptase polymerase chain reaction measurement of peroxisome proliferator-activated receptor-gamma (PPARgamma) and lipoprotein lipase (LPL) were measured. Antioxidant glutathione levels (GSH) and glutathione peroxidase activity (GPX) were determined.


RESULTS
Sod2(-/-) cells demonstrated constitutive adipocytogenesis in basal medium and generated 34% more adipocytes in adipocytogenic media. Growth of cells in the free radical scavenger antioxidant, amifostine (WR2721; 4 mM) decreased numbers of adipocytes in Sod2(-/-) BMSCs in both basal (38.0%, p = 0.037) and adipocytogenic (37.5%, p = 0.021) media and reduced to undetectable the levels of expression of PPARgamma and LPL. In contrast, Sod2(+/+) cells showed no detectable constitutive adipocytogenesis but formed adipocytes in adipocytogenic medium, with a decrease (43.7%, p = 0.001) by addition of WR2721. In basal conditions, Sod2(-/-) cells had lower GSH (78.6%; p = 0.0089) and GPX (52.7%; p < 0.001) levels than did Sod2(+/+) cells, which were increased in either medium by WR2721 treatment of Sod2(-/-) or Sod2(+/+) cells (all p < 0.001). Differentiation of BMSCs to adipocytes was inversely correlated with the level of GSH (r = -0.9427, p = 0.0167). Sod2(-/-) long-term bone marrow cultures had decreased hematopoiesis compared to those from Sod2(+/-) or Sod2(+/+) mice.


CONCLUSIONS
The cellular redox pathway has a role in adipocyte differentiation of cells of the hematopoietic microenvironment.
Lechpammer
Stanislav
S
Department of Orthopedic Surgery, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
Epperly
Michael W
MW
Zhou
Shuanhu
S
Nie
Suhua
S
Glowacki
Julie
J
Greenberger
Joel S
JS
D017667:Adipocytes / Q000166:cytology / Q000502:physiology*; D004999:Amifostine / Q000494:pharmacology; D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D005978:Glutathione / Q000378:metabolism*; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D010084:Oxidation-Reduction / Q000187:drug effects; D047495:PPAR gamma / Q000096:biosynthesis / Q000235:genetics; D011837:Radiation-Protective Agents / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000099156:Superoxide Dismutase 2
D004999:Amifostine; D000975:Antioxidants; D005978:Glutathione; D008071:Lipoprotein Lipase; D047495:PPAR gamma; D011837:Radiation-Protective Agents; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2