LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro
Abstract Background This study was aimed to evaluate the effect of LPS in glucocorticoid receptor (GR) isoforms expression on different cell lines and PBMC from healthy donors in vitro and glucocorticoid sensitivity of PBMC in vitro. Methods U-2 OS cell lines expressing GR isoforms, different cell l...
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| Format: | Article |
| Language: | English |
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BMC
2017-10-01
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| Series: | Journal of Inflammation |
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| Online Access: | http://link.springer.com/article/10.1186/s12950-017-0169-0 |
| _version_ | 1819054188775079936 |
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| author | Maria Luisa Molina Julia Guerrero John A. Cidlowski Héctor Gatica Annelise Goecke |
| author_facet | Maria Luisa Molina Julia Guerrero John A. Cidlowski Héctor Gatica Annelise Goecke |
| author_sort | Maria Luisa Molina |
| collection | DOAJ |
| description | Abstract Background This study was aimed to evaluate the effect of LPS in glucocorticoid receptor (GR) isoforms expression on different cell lines and PBMC from healthy donors in vitro and glucocorticoid sensitivity of PBMC in vitro. Methods U-2 OS cell lines expressing GR isoforms, different cell lines (CEM, RAJI, K562 and HeLa) or PBMC from healthy donors, were cultured or not with LPS. The expression of GRα and GRβ was evaluated by Western blot. Glucocorticoid sensitivity was evaluated in PBMC treated with LPS, testing genes which are transactivated or transrepressed by glucocorticoid. For transactivated genes (MKP1, FKBP5) PBMC were treated with Dexamethasone 100 nM for 6 h. The mRNA expression was measured by RT-PCR. For transrepressed genes (IL-8, GM-CSF), PBMC were cultured in Dexamethasone 100 nM and LPS 10 μg/ml for 6 h and protein expression was measure by ELISA. Results GR isoforms were induced in U-2 OS cells with a greater effect on GRα expression. Both isoforms were also induced in CEM cells with a tendency to a greater effect on GRβ. LPS induced only the expression of GRα in Raji and HeLa cells, and in PBMC, with no effect in K562 cells. LPS induced a loss of glucocorticoid inhibitory effect only on the secretion of GM-CSF. Conclusion LPS in vitro differentially modulates the expression of GR isoforms in a cell specific manner. In PBMC from healthy donors LPS induces an approximately two times increase in the expression of GRα and a loss of the glucocorticoid inhibitory effect on the secretion of GM-CSF, without affecting other glucocorticoid responses evaluated. |
| first_indexed | 2024-12-21T12:47:39Z |
| format | Article |
| id | doaj.art-59b517a03ef84f13ad1ace0abfc4dcc6 |
| institution | Directory Open Access Journal |
| issn | 1476-9255 |
| language | English |
| last_indexed | 2024-12-21T12:47:39Z |
| publishDate | 2017-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Inflammation |
| spelling | doaj.art-59b517a03ef84f13ad1ace0abfc4dcc62022-12-21T19:03:35ZengBMCJournal of Inflammation1476-92552017-10-011411810.1186/s12950-017-0169-0LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitroMaria Luisa Molina0Julia Guerrero1John A. Cidlowski2Héctor Gatica3Annelise Goecke4Rheumatology Section, Internal Medicine Department, Clinical Hospital, University of ChilePhysiology and Biophysics Disciplinary Program, ICBM, Faculty of Medicine, University of ChileDepartment of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of HealthRheumatology Section, Internal Medicine Department, Clinical Hospital, University of ChileRheumatology Section, Internal Medicine Department, Clinical Hospital, University of ChileAbstract Background This study was aimed to evaluate the effect of LPS in glucocorticoid receptor (GR) isoforms expression on different cell lines and PBMC from healthy donors in vitro and glucocorticoid sensitivity of PBMC in vitro. Methods U-2 OS cell lines expressing GR isoforms, different cell lines (CEM, RAJI, K562 and HeLa) or PBMC from healthy donors, were cultured or not with LPS. The expression of GRα and GRβ was evaluated by Western blot. Glucocorticoid sensitivity was evaluated in PBMC treated with LPS, testing genes which are transactivated or transrepressed by glucocorticoid. For transactivated genes (MKP1, FKBP5) PBMC were treated with Dexamethasone 100 nM for 6 h. The mRNA expression was measured by RT-PCR. For transrepressed genes (IL-8, GM-CSF), PBMC were cultured in Dexamethasone 100 nM and LPS 10 μg/ml for 6 h and protein expression was measure by ELISA. Results GR isoforms were induced in U-2 OS cells with a greater effect on GRα expression. Both isoforms were also induced in CEM cells with a tendency to a greater effect on GRβ. LPS induced only the expression of GRα in Raji and HeLa cells, and in PBMC, with no effect in K562 cells. LPS induced a loss of glucocorticoid inhibitory effect only on the secretion of GM-CSF. Conclusion LPS in vitro differentially modulates the expression of GR isoforms in a cell specific manner. In PBMC from healthy donors LPS induces an approximately two times increase in the expression of GRα and a loss of the glucocorticoid inhibitory effect on the secretion of GM-CSF, without affecting other glucocorticoid responses evaluated.http://link.springer.com/article/10.1186/s12950-017-0169-0LipopolysaccharideGlucocorticoid receptorGlucocorticoid resistanceAcute inflammation |
| spellingShingle | Maria Luisa Molina Julia Guerrero John A. Cidlowski Héctor Gatica Annelise Goecke LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro Journal of Inflammation Lipopolysaccharide Glucocorticoid receptor Glucocorticoid resistance Acute inflammation |
| title | LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| title_full | LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| title_fullStr | LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| title_full_unstemmed | LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| title_short | LPS regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| title_sort | lps regulates the expression of glucocorticoid receptor α and β isoforms and induces a selective glucocorticoid resistance in vitro |
| topic | Lipopolysaccharide Glucocorticoid receptor Glucocorticoid resistance Acute inflammation |
| url | http://link.springer.com/article/10.1186/s12950-017-0169-0 |
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