Beneficial effects of Heqi san on rat model of polycystic ovary syndrome through the PI3K/AKT pathway - PubMed (original) (raw)

Beneficial effects of Heqi san on rat model of polycystic ovary syndrome through the PI3K/AKT pathway

Hengxia Zhao et al. Daru. 2017.

Abstract

Background: Heqi San, a traditional Chinese medicine (TCM) has been reported to regulate hormone levels in patients with metabolic disease, suggesting a potential clinical application. In the current study, we aimed to elucidate the effect of Heqi San on rat model of polycystic ovary syndrome (PCOS).

Method: PCOS model was established in female SD rats. Rats were randomly divided into four groups: the control, untreated PCOS model, Heqi San treated PCOS model (8.1 g/kg) and metformin (MET) treated PCOS model (135 mg/kg) groups. All animals were subcutaneously injected with 6 mg/100 g dehydroepiandrosterone (DHEA) in the neck once a day for 20 consecutive days. The serum hormone levels were measured by ELISA. The ovarian tissues were stained with hematoxylin and eosin (HE) to undergo pathological examination. The expression levels of GLTU4 and PTEN mRNA were examined by real time PCR. The crucial proteins in the PI3K/APT pathway were analyzed by western blotting. Then, the functions of the target genes were analyzed using bioinformatics approaches.

Results: We found that Heqi San was able to recover the serum hormone levels and improve insulin resistance in PCOS rat model. A morphological lesion of the ovary was also restored with the Heqi San treatment. More importantly, we discovered a correlation between the PI3K/AKT signaling pathway and the beneficial effects of Heqi San, demonstrating that its application could alter the expression levels of p-ERK, p-AKT, p-GSK3β, IRS-1, PTEN and GLTU4, all key factors in the PI3K/APT pathway. Through a bioinformatical analysis, we predicted the related gene function and pathway of the pathological mechanism of PCOS and found miRNAs that are likely to be critical in PCOS occurrence, including rno-miR-144-3p, rno-miR-30c-2-3p, rno-miR-486, rno-miR-3586-3p and rno-miR-146b-5p.

Conclusion: The beneficial effects of Heqi on PCOS, including alter serum hormone levels, recover ovary morphological lesions and improve insulin resistance, which is mediated through the PI3K/AKT pathway. The potential role of miRNA-144-3p in PCOS pathogenesis.

Keywords: Heqi san; PI3K/AKT pathway; Polycystic ovary syndrome; miRNA PTEN.

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Conflict of interest statement

Ethics approval

The animal study was approved by the Animal Ethics Committee of the Guangzhou University of Chinese Medicine.

All authors agree to publish our manuscript.

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The authors have declared no conflict of interest.

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Figures

Fig. 1

Fig. 1

Establishment of PCOS model. a Testosterone concentration in serum was increased in PCOS model. b FINS level was elevated in PCOS model. *p < 0.05 vs. control

Fig. 2

Fig. 2

Application of Heqi San restored serum hormone levels in the PCOS model, including (a) luteinizing hormone (LH), (b) Testosterone (T), (c) estradiol (E2), and (e) follicule-stimulating hormone (FSH), but not (e) progesterone (P). *p < 0.05 vs. control, # p < 0.05 vs. PCOS

Fig. 3

Fig. 3

Heqi San treatment changed the insulin sensitivity in the PCOS model. a HOMA-IR was alleviated in both Heqi San-treated and MET-treated PCOS model. b Insulin sensitivity index (ISI) was decreased in the PCOS model, while Heqi San counteracted this effect. Heqi: Heqi San, MET: metformin. *p < 0.05 vs. control, # p < 0.05 vs. PCOS

Fig. 4

Fig. 4

Morphological alteration in the ovary after Heqi San treatment. a Ovarian volume (mm3) was slightly changed after Heqi San treatment. b Organ coefficiency (mg/g) improved after Heqi San treatment in the PCOS model. c Compared with the control, the PCOS model showed apparent lesions in the ovary, Heqi San or MET treatment significantly restored the morphological damage in the PCOS model. Heqi: Heqi San, MET: metformin

Fig. 5

Fig. 5

The effects of Heqi San on the PCOS model was mediated through the PI3K/AKT pathway. Heqi San treatment increased the expression levels of GLUT4 (a) or PTEN (b) in the PCOS model, where both of key signals were significantly down-regulated. Heqi: Heqi San, MET: metformin. *p < 0.05 vs. control, # p < 0.05 vs. PCOS

Fig. 6

Fig. 6

Insulin resistance was changed by Heqi San treatment in the PCOS model. a Treatment with either Heqi San or MET significantly improved the response to insulin stimulation, leading to the up-regulation of p-ERK, p-AKT, or GSK3β in the PCOS model. b Heqi San treatment recovered the decrease in IRS-1 and PTEN expression levels, which were inhibited in the PCOS model. Heqi: Heqi San, MET: metformin

Fig. 7

Fig. 7

Analysis of the potential roles of miRNAs in PCOS pathogenesis. a The potential role of rno-miR-144-3p, the target gene of PTEN, was elucidated through real time PCR in the PCOS model and the Heqi-treated group. ** p < 0.01 vs. control, # p < 0.05 vs. PCOS. b GO analysis disclosed the functions of target genes. c GO enrichment analysis demonstrated the signal pathways potentially involved in pathological mechanism of PCOS. Heqi: Heqi San, MET: metformin

Fig. 8

Fig. 8

Differential expression levels of miRNA participating in PCOS pathogenesis, including miR-30c-2-3p (a), miR-146b-5p (b), miR-486 (c), and miR-3586-3p. **p < 0.01 vs. control, # p < 0.05 vs. PCOS

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