[Effect of Bushen Tiaojing Recipe containing serum on FSH/cAMP-PKA pathway in in vitro cultured human ovarian granular cells].
OBJECTIVE
To explore the potential molecular mechanisms for Bushen Tiaojing Recipe (BTR) improving the endocrine function of ovarian granular cells by observing the effect of BTR containing serum on follicle stimulating hormone/cyclic adenosine monophosphate-protein kinase A (FSH/ cAMP-PKA) pathway in in vitro cultured human ovarian granular cells.


METHODS
The primary ovarian granular cells collected from in vitro fertilization-embryo transfer patients were cultured for 24 h. The human and rat serum containing different concentrations of BTR (low, medium, high dose), and their normal serums were co-incubated with ovarian granular cells for 48 h respectively, and then they were divided into the low, medium, high dose BTR groups and the control group. The levels of estradiol (E2), progesterone (P), and cyclic adenosine monophosphate (cAMP) in the culture medium were measured by radioimmunoassay. The protein expression of FSHR in ovarian granular cells was detected by Western Blot. The mRNA expression of follicle stimulating hormone receptor (FSHR) and P450 aromatase (P450arom) in ovarian granular cells were detected by Real-time PCR.


RESULTS
In human BTR containing serum groups: Compared with control group, the levels of E2 and cAMP in the culture medium were higher (both P < 0.05) in the medium and high dose BTR groups; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells increased (all P < 0.01), the mRNA expressions of P450arom in ovarian granular cells were higher (P < 0.05, P< 0.01) in the medium and high dose BTR groups. In rat BTR containing serum groups: Compared with the control group, the levels of E2 in the culture medium were higher (all P < 0.01), cAMP in the culture medium were higher (P < 0.05, P < 0.01) in the medium and high dose BTR group; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells were higher (all P < 0.01), the mRNA expression of P450arom in ovarian granular cells increased in the medium and high dose BTR groups (P < 0.05, P < 0.01).


CONCLUSIONS
BTR could possibly improve the endocrine function of ovarian granular cells by regulating main effector molecules FSHR, cAMP, P450arom, and E2 in FSH/cAMP-PKA pathway of ovarian granular cells.
Cui-Miao
CM
Song
Hong-Min
HM
Yang
Jun
J
Lu
Nan
N
Zhou
Shuai
S
Lu
Yan-Cang
YC
Duan
Hui-Rong
HR
Ma
Hua-Zhou
HZ
Xu
Hui-Lan
HL
Du
D002478:Cells, Cultured; D054743:Cyclic AMP-Dependent Protein Kinase Type I / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D044967:Serum / Q000737:chemistry; D015398:Signal Transduction / Q000187:drug effects
D004365:Drugs, Chinese Herbal; D005640:Follicle Stimulating Hormone; D054743:Cyclic AMP-Dependent Protein Kinase Type I