Table 1 Treated animal grouping and description.

I

Normal control

Mice were fed a normal diet and 3 mL of distilled water orally for 8 weeks

II

PCOS-induced group

[HFD + Letrozole (90 µg /kg.b.w.)]

Received HFD for 8 weeks²¹ and given letrozole by gavage (90 µg/kg.b.w) daily for the last 3 weeks²². The dosage of letrozole (90 µg/kg b.w.) used to induce PCOS in mice is based on the protocol established by Kafali et al.,²²which reliably reproduces PCOS-like ovarian cysts and hormonal disturbances in rodents.

III

PCOS + MTF

(500 mg/kg.b.w)

Given HFD for 8 weeks + Letrozole by gavage (90 µg/ kg.b.w) + MTF (500 mg/ kg. B.W.) suspended in distilled water daily along with letrozole for the last 3 weeks.

IV

PCOS + MTL

(500 mg/kg.b.w)

Given HFD for 8 weeks + Letrozole by gavage (90 µg/ kg.b.w) + MTL (500 mg/ kg. B.W.) suspended in distilled water daily along with letrozole for the last 3 weeks.

V

PCOS + MVF

(500 mg/kg.b.w)

Given HFD for 8 weeks + Letrozole by gavage (90 µg/ kg.b.w) + MVF (500 mg/ kg. B.W.) suspended in distilled water daily along with letrozole for the last 3 weeks.

VI

PCOS + MVL

(500 mg/kg.b.w)

Given HFD for 8 weeks + Letrozole by gavage (90 µg/ kg.b.w) + MVL (500 mg/ kg. B.W.) suspended in distilled water daily along with letrozole for the last 3 weeks.

VII

PCOS + Metformin

(300 mg/kg.b.w)

Given HFD for 8 weeks + Letrozole by gavage (90 µg/ kg.b.w) + Metformin (300 mg/kg b.w) suspended in distilled water daily along with letrozole for the last 3 weeks. The selected dose of metformin (500 mg/kg b.w.) was based on prior in vivo studies demonstrating therapeutic efficacy in improving hormonal, metabolic, and ovarian dysfunctions in PCOS animal models without inducing systemic toxicity. A comparable dose was used by Di Pietro et al. (2015)²³who reported significant modulation of ovarian angiogenesis and follicular development using metformin in a letrozole-induced PCOS rat model.