Polycystic ovary syndrome (PCOS), a common hormonal disorder in women of reproductive age, is associated with a poor and unhealthy diet. This study aimed to investigate the effect of a high sucrose and cholic acid (HSCA) diet in the presence of PCOS-like phenotypes. Female Wistar rats were divided into HSCA and normal diet groups for four weeks, each with twenty rats. Body weight was assessed before and after the study. Blood and fecal samples were obtained to measure HOMA-IR and testosterone level (ELISA) and Enterobacteriaceae isolates grown on MacConkey Agar. Obtained ovarian tissues were H&E-stained. HSCA rats demonstrated a reduction in Enterobacteriaceae colonies (median 4.75 × 105 vs. 2.47 × 104/CFU, p < 0.001) and an elevated HOMA-IR (mean 2.94 ± 1.30 vs. 4.92 ± 0.51, p < 0.001), as well as an increase in testosterone level (median 0.65 vs. 3.00 ng/mL, p < 0.001), despite no statistical differences in the change in body weight (mean −2.31 ± 14.42 vs. −3.45 ± 9.32, p = 0.769). In H&E staining, HSCA rats had a reduction in preovulatory follicle count (median 0.50 vs. 0.00, p = 0.005). The HSCA diet caused insulin resistance and high testosterone levels, which contribute to the development of PCOS, and affected folliculogenesis by altering follicular maturation, but had no effect on ovulation.