The effects of Sutaehwan-Gami on menopausal symptoms induced by ovariectomy in rats

Abstract

Background: This study was undertaken to evaluate the beneficial effects of a modified prescription of Sutaehwan named Sutaehwan-Gami (SG), created by adding Rhizoma dioscoreae and Carthami semen to Sutaehwan, on menopausal symptoms.

Methods: To evaluate the estrogenic effect of SG, we first examined estrogen receptor (ER) activation by SG treatment in breast adenocarcinoma cells and confirmed the estrogenic effect of SG in vivo ovariectomized rats. The animals were randomized into four groups: Sham operated group (Sham), saline treated ovariectomized group (OVX), SG treated group (SG) and raloxifene treated group (RLX). Animals were provided with SG at a dose of 500 mg/kg bw/day and RLX at a dose of 5.4 mg/kg bw/day with standard rat pellets for 3 months.

Results: SG significantly increased ERα phosphorylation, and its downstream effectors, extracellular signal-regulated kinase (ERK) and protein kinase B (Akt) phosphorylation in breast adenocarcinoma cells. Treatment with SG reversed ovariectomy-induced uterine weight reduction and weight gain. Decreases in the levels of GOT and GPT were observed in the SG group. The significantly reduced E2β level in OVX rats was raised by treatment with SG. Moreover, SG significantly increased the phosphorylation levels of ERK and Akt in the uterus.

Conclusion: Taken together, these data indicate that SG has phytoestrogen-like properties through ERK and Akt activation, implying that it could be protective and beneficial for the management of menopausal symptoms.

Figure 1

Effect of SG on cell viability. MCF-7 cells were treated with different concentrations of SG for 48 hours. Cell viability was determined by MTT assay. Data are mean ± SE of three experiments performed in triplicate. *P < 0.05 versus 0 μg/ml.

Figure 2

Effect of SG on the activation of ERα, ERK and Akt in the MCF-7 cells. Total and phosphorylation state of each protein was measured in MCF-7 cells which were treated with SG at 1 μg/ml for the indicated time. The whole cell lysates were immunoblotted with anti-p- ERα/ERK/Akt or ERα/ERK/Akt antibody. The intensity of the phosphorylated or total ERα/ERK/Akt was quantified by densitometry, and the intensity of the phosphorylated ERα/ERK/Akt band was normalized to that of total ERα/ERK/Akt. *P < 0.05 versus control.

Figure 3

Effect of SG on the activation of ERK and Akt. Ovariectomized rats were treated with SG or raloxifene for 3 months. The uterine tissue lysates were immunoblotted with anti-p-ERK, anti p-Akt, ERK antibody and Akt antibody. The intensity of protein was quantified by densitometry and the intensity of the phosphorylated ERK or Akt band was normalized to that of total ERK or total Akt, respectively. †P < 0.05 versus OVX.

Figure 4

Effect of SG on uterine morphology in ovariectomized rats. Following three month treatment, animals were sacrificed and uterus was removed. Formalin-fixed rat uteri were paraffin-sectioned serially into 4 μm sections to be stained with hematoxylin and eosin. A: Sham, B: OVX, C: SG, D: RLX, Bar = 500 μm.