Testosterone

Hyaluronic acid (HA), methacrylic anhydride, 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone, poly (vinyl alcohol) (PVA), and Minoxidil were all purchased from Aladdin Bio-Chem Technology Co., Ltd (Shanghai, China). Poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid-co-3-hydroxyhexanoic acid trimer) (PHBVHHx or PBVHx), a novel trimer of PHA containing 90 mol% of 3-hydroxybutyrate, 3 mol% of 3-hydroxyvalerate and 7 mol% of 3-hydroxyhexanoate purchased from Bluepha Co., Ltd (Beijing, China). Testosterone was purchased from Solarbio Science & Technology Co., Ltd. (Shanghai, China). Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Laboratories (Kumanoto, Japan). Calcein acetoxymethyl ester (Calcein-AM) and 3,8-diamino-5-[3-(diethylmethylammonio) propyl]-6-phenylphenanthridinium diiodide (PI), crystal violet staining solution, matrix-Gel™ (growth factor reduced) were obtained from Beyotime Biotechnology (Shanghai, China). VEGF and VEGFA ELISA Kit were purchased from ABclonal Technology Co., Ltd.

The following are used in the study: testosterone, paracetamol, phenacetin, and NADPH (Beijing Solarbio Science & Technology Co., Ltd. China); dextran, dextromethorphan, omeprazole, 5-hydroxy omeprazole, tolbutamide, 4-hydroxytolbutamide, and 6β-hydroxytestosterone (Toronto Research Chemicals); BCA protein Kit (Lianke Biotechnology Corporate Limited, No. a81911151); primer (synthesized by Tsingke Biotechnology Co., Ltd.); reverse transcription Kit (Tsingke Biotechnology Co., Ltd., No. 20190328); and CYP1A2, cyp2d1, CYP3A4, CYP3A1, CYP2C19, and CYP2C11 antibodies (Abcam Company).

The primary granulosa cells were seeded onto 6-well plates at a density of 1 × 10⁶ cells per well, conventionally cultured for 24 h, starved in the starvation culture medium for 24 h, and then treated with the corresponding concentrations of Baicalin with 10^− 7 M testosterone (Solarbio) added simultaneously to the medium as the substrate for estradiol (E₂) production. After 48 h of treatment, the cell culture supernatant was collected, and the concentrations of estradiol and progesterone were analyzed using a chemiluminescent immunoassay kit (Roche Diagnostics, Rotkreuz, Switzerland) and cobas 6000 analyzer series (Roche Diagnostics) according to the kit manufacturer’s instructions.