Vancomycin hydrochloride

The fluorophore NHS esters were purchased from Okeanos Technology (Beijing, China). Vancomycin hydrochloride, metronidazole, polymyxin B sulfate, and cefotaxime sodium were bought from Sangon Biotech (Shanghai, China). And other chemicals, not noted above, were from Sigma-Aldrich (St. Louis, MO, USA).

Eighteen rats were randomly allocated to 3 groups: PBS, DHEA and DHEA + antibiotics groups (n = 6 each group). To induce the PCOS phenotype, rats in the DHEA group were subcutaneously injected with 6 mg/100 g bodyweight DHEA (cat. no. H10940064, Yangzhou Pharmaceutical Co., Ltd) daily for 35 consecutive days. Rats in the PBS group were injected with PBS. Rats in the DHEA + antibiotics group were injected with DHEA in the same manner as the DHEA group and received a freshly prepared antibiotic cocktail daily in their drinking water. The antibiotic cocktail, consisting of ampicillin sodium (1 mg mL⁻¹), neomycin sulfate (1 mg mL⁻¹), metronidazole (1 mg mL⁻¹), and vancomycin hydrochloride (0.5 mg mL⁻¹) (all purchased from Sangon Biotech, China) [54 (link)], was administered to rats every day immediately after preparation. The rats in all 3 groups were sacrificed after 5 weeks of treatment.

The antibodies used in this study were as follows: TH (Sigma Aldrich, AB152) and GAPDH (KeyGEN, KGAA002). The reagents used for western blot were all obtained from Sigma Aldrich. The remaining reagents used in the animal study included osteocalcin (Bachem, H-6552), sodium propionate (Sigma Aldrich, P1880), sodium chloride (Sigma Aldrich, S3014), 6-OHDA (Sigma Aldrich, H116), desipramine (Sigma Aldrich, D3900), ampicillin, neomycin, metronidazole, and vancomycin hydrochloride were all purchased from Sangon Biotech. The BCA kit was obtained from Thermo Fisher Scientific (23259), the QIAamp DNA Stool Mini Kit was purchased from QIAGEN (51504), and the GLP-1 ELISA kit was from Millipore (EGLP-35 K). And the ABC kit and DAB kit were purchased from Vectastain Elite.