Steroidogenic effects of Taraxacum officinale extract on the levels of steroidogenic enzymes in mouse Leydig cells - PubMed (original) (raw)

Steroidogenic effects of Taraxacum officinale extract on the levels of steroidogenic enzymes in mouse Leydig cells

Hyun Joo Chung et al. Anim Cells Syst (Seoul). 2018.

Abstract

In this study, we investigated the steroidogenic effect of Taraxacum officinale extract on mouse TM3 Leydig cells, which produce male hormones by increasing the levels of steroidogenic enzymes. Steroidogenic enzymes are involved in the production of testosterone in the testis. To date, the steroidogenic effect of T. officinale has not been reported. Therefore, we examined the steroidogenic effects of T. officinale extract (TOE) on mouse Leydig cells in vitro. Traditionally, plants have been used for the treatment of various kinds of ailments. For many years, some medicinal plants have been used to regulate steroidogenesis or late-onset hypogonadism (LOH). In particular, plants belonging to the genus Taraxacum have anti-inflammatory, anti-nociceptive, anti-oxidant, and anti-cancer properties. In this study, we determined whether the TOE exerts steroidogenic effects by increasing the levels of enzymes associated with steroidogenesis, such as the steroidogenic acute regulatory protein (STAR), CYP11A1, and translocator protein (TSPO) in the mitochondria and CYP17A1 in the smooth endoplasmic reticulum, in mouse Leydig cells. Our results showed that the TOE significantly increased the mRNA and protein levels of steroidogenic enzymes, thereby increasing the testosterone levels in mouse Leydig cells. Thus, our results indicate that the TOE increases the levels of steroidogenic enzymes, and further studies are required to establish the potential of this plant in regulating steroidogenesis and improving LOH.

Keywords: Leydig cell; Taraxacum officinale extract (TOE); late-onset hypogonadism (LOH); medicinal plant; steroidogenesis.

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Figures

Figure 1.

Figure 1.

HPLC chromatograms of standard sample (A) and Taraxacum officinale extract (B). Peak1, Chicoric acid. The compound in T. officinale was identified on the basis of the retention time and chromatogram pattern, by co-chromatography with an authentic standard. From the data presented, peak 1 with a retention time of 18.45 min, was chicoric acid. The contents of chicoric acid in our study had high level of around 2.6 mg/g.

Figure 2.

Figure 2.

Cell viability of TM3 Leydig cells treated Taraxacum officinale extract. The varying concentration of 0, 1, 10, 25 and 50 µg/ml of T. officinale were treated into TM3 cells for 24(A) and 48 h(B). The experiments were performed in triplicate.

Figure 3.

Figure 3.

Measurement of mRNA levels of Cyp11a1, Cyp17a1 and StAR in mouse Leydig cells. Cells were treated with 0, 1, 10, 25 and 50 µg/ml of T. officinale for 24 h. The qRT-PCR was performed with the gene-specific primers for Cyp11a1 (A), Cyp17a1 (B) and StAR (C). mRNA were normalized with 18s rRNA gene. The experiments were performed in triplicate.

Figure 4.

Figure 4.

Measurement of protein levels of CYP11A1, CYP17A1 and STAR in mouse TM3 Leydig cells. Cells were treated with 0, 1, 10, 25 and 50 µg/ml of T. officinale for 48 h. The Western blotting analysis was performed with the gene-specific antibodies for CYP11A1, CYP17A1 and STAR (A). Total protein were normalized with ß-ACTIN. The data were quantified by using Image J program (B). The experiments were performed in triplicate.

Figure 5.

Figure 5.

Measurement of levels of testosterone in mouse Leydig cell supernatants. Cells were treated with 0, 1, 10, 25 and 50 µg/ml of T. officinale for 48 h. The cell supernatant were collected and used to do the ELISA method. Each group's _p_-value was calculated. (*p < .05, **p < .001). The experiments were performed in three times, respectively.

Figure 6.

Figure 6.

The model of steroidogenesis with the treatment of TOE in mouse Leydig cell.

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Grants and funding

This work was supported by a grant [714001-07] from the Center for Industrialization of Natural Nutraceuticals through the Agriculture, Food and Rural Affairs Research Center Support Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea. Also this research was supported by the Chung-Ang University Research Scholarship Grants in 2017.

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