Direct rosiglitazone action on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells.
BACKGROUND
Ovarian granulosa cells are the predominant source of estradiol and progesterone biosynthesis in vivo. Rosiglitazone, a synthetic agonist of the peroxisome proliferator-activated receptor gamma (PPAR gamma), is applied as the treatment of insulin resistance including women with PCOS. The aim of the study was to investigate the direct effects of rosiglitazone on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells (GLCs).


METHODS
Primary human GLCs were separated during in vitro fertilization and cultured in the presence of rosiglitazone, GW9662 (an antagonist of PPAR gamma) and hCG. The mRNA expression of key steroidogenic factors including 3beta- hydroxysteriod dehydrogenase (3beta-HSD), cytochrome P-450 scc (CYP11A1), cytochrome P-450 aromatase (CYP19A1), and steroidogenic acute regulatory protein (StAR) were detected by quantitative real-time PCR. Estradiol and progesterone levels in GLCs cultures were measured by chemiluminescence immunoassay, and the proinflammtory factors (TNFalpha and IL-6) in conditioned culture media were measured by ELISA.


RESULTS
PPAR gamma mRNA levels increased up to 3.24 fold by rosiglitazone at the concentration of 30 microM compared to control (P<0.05). hCG alone or hCG with rosiglitazone had no significant effects on PPAR gamma mRNA levels. The CYP19A1 mRNA level at exposure to rosiglitazone alone showed a drop, but was not significantly reduced comparing to control. The expression levels of enzymes 3beta-HSD and CYP11A1 in all treatments did not alter significantly. The StAR mRNA expression at exposure to rosiglitazone was significantly increased comparing to control (P<0.05). The media concentrations of E2 and progesterone by rosiglitazone treatment showed a declining trend comparing to control or cotreatment with hCG, which did not reach significance. Most importantly, treatment with rosiglitazone decreased TNFalpha secretion in a statistically significant manner compared with control (P<0.05). The concentration of IL-6 following rosiglitazone exposure did not significantly decrease comparing to control.


CONCLUSIONS
In cultured GLCs, rosiglitazone stimulated StAR expression, but did not significantly affect steroidogenic enzymes, as well as E2 and progesterone production. Moreover, rosiglitazone significantly decreased the production of TNFalpha in human GLCs, suggesting that PPAR gamma may play a role in the regulation of GLCs functions through inhibiting proinflammatory factors.
Chen
Qiuju
Q
NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai, PR China. chenqiuju2865@hotmail.com
Sun
Xiaoxi
X
Chen
Junling
J
Cheng
Linan
L
Wang
Jian
J
Wang
Yongwei
Y
Sun
Zhaogui
Z
D000328:Adult; D000813:Anilides / Q000494:pharmacology; D006063:Chorionic Gonadotropin / Q000096:biosynthesis; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000166:cytology; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis*; D008184:Luteal Cells / Q000187:drug effects / Q000378:metabolism*; D047495:PPAR gamma / Q000819:agonists*; D011374:Progesterone / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077154:Rosiglitazone; D013256:Steroids / Q000096:biosynthesis*; D045162:Thiazolidinediones / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
C457499:2-chloro-5-nitrobenzanilide; D000813:Anilides; D006063:Chorionic Gonadotropin; D015850:Interleukin-6; D047495:PPAR gamma; D013256:Steroids; D045162:Thiazolidinediones; D014409:Tumor Necrosis Factor-alpha; D000077154:Rosiglitazone; D011374:Progesterone; D004958:Estradiol; D012313:RNA