Lobetyolin Reshapes Gut Microbiota and Bile Acid Metabolism to Attenuate Androgen-Driven PCOS Phenotypes in Mice

To examine lobetyolin (LT)'s integrated regulatory role and molecular mechanisms in polycystic ovarian syndrome (PCOS)'s reproduction, metabolism, inflammation, and uterine receptivity. Methods: Dehydroepiandrosterone (DHEA) was used to create a mouse model of polycystic ovarian syndrome (PCOS). Lobetyolin was administered to the mice via intraperitoneal injection (i.p.) daily for 28 consecutive days. Monitoring estrus cycle, glucose tolerance, and hormone levels. ELISA, qPCR, 16S rRNA sequencing, and targeted bile acid metabolomics were used to assess the expression of androgen synthesis genes (cholesterol side-chain cleavage enzyme, CYP11A1; 17α-Hydroxylase/17,20-lyase, CYP17A1; Aromatase, CYP19A1), inflammation-microbiota-bile acid axis, and endometrial matrix metalloproteinases (MMPs). Lobetyolin effectively restored the estrus cycle, lowered the LH/FSH ratio and free testosterone levels (P < 0.01), and enhanced insulin sensitivity in DHEA mice. LT downregulated CYP11A1 and CYP17A1 and upregulated CYP19A1 via "upstream inhibition and downstream activation" in the androgen network. Additionally, it reduced ovarian IL-6 and TNF-α levels by inhibiting the TLR4/NF-κB pathway. LT altered the gut microbiota by increasing the α-diversity index, normalizing the Firmicutes/Bacteroidetes (F/B) ratio, promoting Dubosiella and Muribaculum growth, and inhibiting Lachnospiraceae and Alistipes abundance. HCA and TCDCA levels were specifically enhanced in the bile acid profile by LT. LT improves endometrial receptivity by reducing VEGFA/VEGFR expression, upregulating TIMP1 and TIMP2, restoring MMP/TIMP homeostasis, and reversing the "triple network dysregulation" of oxidative stress, inflammation, and vascular remodeling. Conclusion: Lobetyolin exerts therapeutic effects by inhibiting androgen synthesis, insulin sensitization, inflammation, and microbiota remodeling, reprogramming the bile acid profile, and correcting the uterine stroma.” synergistically breaks the PCOS cycle of hyperandrogenism, insulin resistance, chronic inflammation, and defective uterine reprogramming. A novel, exactly translatable, integrated clinical intervention strategy was developed.