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Effects of Bushen Tiaochong Recipe (补肾调冲方) containing serum on ovarian granulosa cell proliferation, steroidogenesis and associated gene expression in rats
  
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KeyWord:serum pharmacology  Bushen Tiaochong Recipe  cell proliferation  steroidogenesis  follicle stimulating hormone receptor  insulin-like growth factor-1
Author NameAffiliationE-mail
Tian Xia The First Clinical Medical College Affiliated to Guangzhou University of TCM, Guangzhou, 510405, China  
Song-ping Luo The First Clinical Medical College Affiliated to Guangzhou University of TCM, Guangzhou, 510405, China  
Yu Fu The First Clinical Medical College Affiliated to Guangzhou University of TCM, Guangzhou, 510405, China  
Bing Han The Second Affi liated Hospital to Tianjin University of TCM, China xiatian76@163.com 
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Abstract:
      Objective To observe the effect of Bushen Tiaochong Recipe (补肾调冲方, BSTCR) on rats’ ovarian granulosa cell (GC) proliferation, steroidogenesis and follicle-stimulating hormone receptor (FSHR), and insulin-like growth factor-1 (IGF-1) mRNA expression using serum pharmacological method. Methods Rats’ GCs were incubated with 10% blank serum (as negative control group), folliclestimulating hormone (FSH)-containing serum (S-FSH, as positive control group), or BSTCR (in different dosages) containing serum (S-BSTCR, as the BSTCR groups) for 48 h. 3H-TdR incorporation was then performed; DNA was measured to analyze the distribution of GCs in the cell cycle and their proliferation index (PI) using a flow cytometer; estradiol (E2) and progesterone (P) content in the culture fl uid were examined by radioimmunoassay; and levels of FSHR and IGF-1 mRNA expression in GCs were measured by real-time RT-PCR. Results A dose-dependent increase of 3H-TdR incorporation in GC was shown in the BSTCR groups. Cells in G0/G1 phase had markedly less, while those in S phase had a signifi cantly higher increase in the BSTCR groups compared with the negative control group. A high value of PI was also shown in the BSTCR groups, especially in the high dose group where the infl uence of cell proliferation was stronger than that in the positive control group. The levels of E2 and P in the BSTCR groups of all dosages were signifi cantly higher than those in the negative control group, and did not show any signifi cant difference compared with those in the positive control group. Levels of FSHR and IGF-1 mRNA expression in the BSTCR groups increased in a dose-dependent manner at levels higher than those in the negative control group. Conclusion S-BSTCR can obviously stimulate the proliferation and steroidogenesis of ovarian GCs. It is speculated that BSTCR could play a regulatory action on ovarian function through two different pathways of endocrine and autocrine by promoting FSHR and IGF-1 mRNA expression. Supported by Jiangsu Provincial Natural Science Foundation (No. BK2005006)
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