Effect of Triptolide on Progesterone Production from Cultured Rat Granulosa Cells

J. Zhang; Z. Jiang; X. Mu; J. Wen; Y. Su; L. Zhang

doi:10.1055/s-0032-1309041

Arzneimittelforschung, Table of Contents

Arzneimittelforschung 2012; 62(06): 301-306
DOI: 10.1055/s-0032-1309041

Original Article

Effect of Triptolide on Progesterone Production from Cultured Rat Granulosa Cells

J. Zhang

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

²The Secondary Affiliated Hospital of Nanjing Medical University, Nanjing, P.R. China

,

Z. Jiang

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

,

X. Mu

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

,

J. Wen

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

,

Y. Su

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

,

L. Zhang

¹Jiangsu Center of Drug Screening, China Pharmaceutical University, Nanjing, P.R. China

› Author Affiliations

Abstract

Triptolide(CAS 38748-32-2), a major active component of Tripterygium wilfordii Hook F (TWHF), is known to have multiple pharmacological activities. However, studies have also shown that triptolide is highly disrupt to the reproductive system by disrupting normal steroid hormone signaling. In the present study, we investigated the effect of triptolide (5, 10, or 20 nM for 24 h) on progesterone production by rat granulosa cells. Triptolide inhibited both basal and human chorionic gonadotropin (HCG)- and 8-bromo-cAMP-stimulated progesterone production as revealed by RIA assay. Furthermore, the HCG-evoked increase in cellular cAMP content was also inhibited by triptolide, indicating that disruption of the cAMP/PKA signaling pathway may mediate the deleterious effects of triptolide on progesterone regulation. In addition, triptolide inhibited 25-OH-cholesterol-stimulated progesterone production, suggesting that activity of the P450 side chain cleavage (P450scc) enzyme was also be inhibited by triptolide. Western blot and quantitative real-time PCR (qRT-PCR) assays further revealed that triptolide decreased mRNA and protein expression of P450scc and the steroidogenic regulatory (StAR) protein in granulosa cells. In contrast, cell viability tests using 3-(4,5-dimethyl-thiazol-2-yl)-2,5- diphenyl-tetrazolium bromide (MTT) indicated that triptolide did not cause measurable cell death at doses that suppressed steroidogenesis. The reproductive toxicity of triptolide may be caused by disruption of cAMP/PKA-mediated expression of a number of progesterone synthesis enzymes or regulatory proteins, leading to reduced progesterone synthesis and reproductive dysfunction.

Key words

triptolide - progesterone - P450scc - StAR - rat granulosa cell

Full Text

References

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