Hypoxia-induced leptin production in human trophoblasts does not protect from apoptosis.
OBJECTIVE
The ob-gene product, leptin, is an important regulator of placental and fetal development during pregnancy. Leptin, being induced by hypoxia in the placenta, is a known pro-apoptotic molecule in adipose tissue but is also known to inhibit apoptosis in other tissues like neuroblastoma cells. Based on these findings, we investigated if leptin has a pro- or anti-apoptotic effect on a trophoblastic cell line (JAr cells) in the presence or absence of oxygen.


RESULTS
Measurement of leptin in the supernatant by using ELISA showed hypoxia-induced leptin production in JAr cells in vitro. This could be confirmed by a leptin-specific RT-PCR. By analyzing leptin and/or hypoxia exposed cells with FACS cytometry we found that JAr cells can cope with hypoxia down to oxygen tensions of 1%. At this level, only a small number of cells underwent apoptosis. Interestingly, leptin added to the culture medium in high concentrations was not able to interfere with the rate of proliferation or apoptosis in these cells independent of the oxygen tension. Finally, an anti-caspase-3 and anti-caspase-9 Western blot was performed. Again, no difference in the expression of caspase-3 and -9 under the conditions tested was seen.


CONCLUSIONS
These results show that leptin, produced by placental cells after hypoxia in vitro, has no influence on the rate of proliferation of these cells. Furthermore, it does not influence apoptotic pathways in the trophoblastic cell line tested under hypoxic and non-hypoxic conditions.
Meissner
Udo
U
Department of Pediatrics, University of Erlangen-Nürnberg, Loschgestrasse 15, Erlangen 91054, Germany. udo.meissner@kinder.imed.uni-erlangen.de
Spranger
Robert
R
Lehner
Manfed
M
Allabauer
Ida
I
Rascher
Wolfgang
W
Dötsch
Jörg
J
D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000096:biosynthesis; D015687:Cell Hypoxia / Q000502:physiology; D006063:Chorionic Gonadotropin / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D020738:Leptin / Q000096:biosynthesis* / Q000235:genetics; D011247:Pregnancy; D020033:Protein Isoforms; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014327:Trophoblasts / Q000378:metabolism*
D006063:Chorionic Gonadotropin; D020738:Leptin; D020033:Protein Isoforms; C505407:CASP3 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases