*Article* **Cannabidiol Decreases Intestinal Inflammation in the Ovariectomized Murine Model of Postmenopause**

**Karen Mae A. Bacalia 1,2, Kevin M. Tveter 1, Hayley Palmer 1, Jeffrey Douyere 1, Savannah Martinez 1, Ke Sui 1 and Diana E. Roopchand 1,\***


**Abstract:** Cannabidiol (CBD) (25 mg/kg peroral) treatment was shown to improve metabolic outcomes in ovariectomized (OVX) mice deficient in 17β-estradiol (E2). Herein, CBD effects on intestinal and hepatic bile acids (BAs) and inflammation were investigated. Following RNA sequencing of colon tissues from vehicle (VEH)- or CBD-treated sham surgery (SS) or OVX mice (*n* = 4 per group), differentially expressed genes (DEGs) were sorted in ShinyGO. Inflammatory response and bile secretion pathways were further analyzed. Colon content and hepatic BAs were quantified by LC-MS (*n* = 8–10 samples/group). Gut organoids were treated with CBD (100, 250, 500 μM) with or without TNFα and lipopolysaccharide (LPS) followed by mRNA extraction and qPCR to assess CBD-induced changes to inflammatory markers. The expression of 78 out of 114 inflammatory response pathway genes were reduced in CBD-treated OVX mice relative to vehicle (VEH)-treated OVX mice. In contrast, 63 of 111 inflammatory response pathway genes were increased in CBD-treated sham surgery (SS) mice compared to VEH-treated SS group and 71 of 121 genes were increased due to ovariectomy. CBD did not alter BA profiles in colon content or liver. CBD repressed *Tnf* and *Nos2* expression in intestinal organoids in a dose-dependent manner. In conclusion, CBD suppressed colonic inflammatory gene expression in E2-deficient mice but was pro-inflammatory in E2-sufficient mice suggesting CBD activity in the intestine is E2-dependent.

**Keywords:** cannabidiol; inflammation; bile acids; transcriptomics; estrogen deficiency; ovariectomized mice; postmenopause; gu<sup>t</sup> organoids
