Bu-shen-zhu-yun decoction promotes synthesis and secretion of FSHβ and LHβ in anterior pituitary cells in vitro

Biomed Pharmacother. 2018 Jun:102:494-501. doi: 10.1016/j.biopha.2018.02.003. Epub 2018 Mar 24.

Abstract

Luteal phase defects (LPD) are an important etiology of infertility which has increased in recent years. Studies have shown that bu-shen-zhu-yun decoction (BSZY-D) can lower the expression of estrogen receptor and progesterone receptor, in rats endometrium of embryonic implantation period, which upregulated by mifepristone, and improve uterine receptivity. The aim of present study was to determine the effect of BSZY-D on the synthesis and secretion of gonadotropic hormones in the anterior pituitary cells of rats. Rats were treated with saline (control) or BSZY-D two times/day for three estrous cycles by gavage. The cerebrospinal fluid (CSF) were collected for further cell treatment. The components in BSZY-D, serum and CSF were analysed by High Performance Liquid Chromatography (HPLC). Cells were either pretreated with normal CSF or BSZY-D/CSF before being stimulated with or without cetrorelix. The mRNA and proteins levels of receptors, hormones, and transcription factors were detected by RT-PCR, western blot analysis and immunostaining. We show that non-toxic concentrations of cetrorelix, a GnRH antagonist, can reduce the mRNA and protein levels of GnRHR, LH, and FSH. This effect could be reversed by the addition of BSZY-D/CSF. We also show decreased mRNA and protein expression of transcription factors, such as CREB, and Egr-1 and secretory vescicles, including SNAP-25 and Munc-18 upon treatment with cetrorelix could be reversed post co-treatment with BSZY-D/CSF. These results indicate that BSZY-D/CSF treatment led to increased levels of GnRHR, transcription factors, and secretory vesicles leading to increased secretion of FSH and LH. Thus, BSZY-D presents a promising candidate to treat luteal phase defects and infertility.

Keywords: Anterior pituitary cells; Bu-shen-zhu-yun decoction; FSH; Infertility; LH.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Chromatography, High Pressure Liquid
  • Drugs, Chinese Herbal / pharmacology*
  • Early Growth Response Protein 1 / metabolism
  • Female
  • Follicle Stimulating Hormone, beta Subunit / biosynthesis*
  • Follicle Stimulating Hormone, beta Subunit / metabolism*
  • Gonadotropin-Releasing Hormone / analogs & derivatives
  • Gonadotropin-Releasing Hormone / pharmacology
  • Luteinizing Hormone, beta Subunit / biosynthesis*
  • Luteinizing Hormone, beta Subunit / metabolism*
  • Munc18 Proteins / metabolism
  • Pituitary Gland, Anterior / cytology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats, Sprague-Dawley
  • Receptors, LHRH / metabolism
  • Synaptosomal-Associated Protein 25 / metabolism
  • Transcription Factors / metabolism
  • Up-Regulation / drug effects

Substances

  • Drugs, Chinese Herbal
  • Early Growth Response Protein 1
  • Egr1 protein, rat
  • Follicle Stimulating Hormone, beta Subunit
  • Luteinizing Hormone, beta Subunit
  • Munc18 Proteins
  • RNA, Messenger
  • Receptors, LHRH
  • Synaptosomal-Associated Protein 25
  • Transcription Factors
  • Gonadotropin-Releasing Hormone
  • cetrorelix