Effect of forskolin on corticosterone secretion in Y1 mouse adrenocortical tumor cells
10.3969/j.issn.1001-1978.2015.10.017
- VernacularTitle:佛司可林促进小鼠肾上腺皮质瘤细胞皮质酮分泌的作用研究
- Author:
Zhiqiang PAN
;
Longlong LIANG
;
Zhaoqin FANG
- Publication Type:Journal Article
- Keywords:
forskolin;
adrenocortical;
Y1 mouse ad-renocortical tumor cell;
corticosterone;
steroidogenic enzyme;
Star;
Cyp11 b1
- From:
Chinese Pharmacological Bulletin
2015;(10):1408-1413
- CountryChina
- Language:Chinese
-
Abstract:
Aim To study the effect of forskolin on cor-ticosterone secretion in mouse adrenocortical tumor cells. Methods Y1 cells were treated with 1μmol· L-1 or 10μmol·L-1 forskolin for 15 min to 24 h. Y1 cells growth morphology was observed, cell culture su-pernatants were collected and corticosterone was tested by ELISA kit. The cells total RNA was extracted using TRIzol kit and was reversely transcribed to obtain the cDNA, then was amplificated by real time quantitative PCR. The cells were lysed with RIPA lysis buffer and protein expressions were carried out by Western blot. Results Y1 cells morphology changed from flat adher-ent to shrink and spherical growth after 1μmol · L-1 forskolin treatment for 3 h. Compared with the control group, corticosterone levels were increased significantly by 1μmol · L-1 forskolin treatment for 24 h ( P <0. 01), then, forskolin significantly enhanced steroid ogenic acute regulatory protein ( Star) mRNA and pro-tein expression ( P <0. 01 ) , moreover, the steroido-genic enzymes such as Cyp11 a1 and Cyp11 b1 mRNA expressions were also up-regulated significantly by fors-kolin ( P <0. 01 ) . Additionally, Star mRNA expres-sion was increased significantly in a time-dependent re-sponse after 10μmol·L-1 forskolin treatment from 1 h to 12 h (P<0. 01). Furthermore, Nr4a1 and Nr4a2 mRNA expressions were up-regulated significantly after 10μmol · L-1 forskolin treatment from 15 min and reached the top at 1 h ( P<0. 01 ) . However, forsko-lin showed no effect on Mc2 r and Nr5 a1 mRNA expres-sions. Conclusion Y1 mouse adrenocortical tumor cells have a strong response to adenylate cyclase ago-nists, then, forskolin can be used to glucocorticoid se-cretion inducer reagent.
