The Bile Acid Synthesis Pathway Is Present and Functional in the Human Ovary

Background: Bile acids, end products of the pathway for cholesterol elimination, are required for dietary lipid and fat-soluble vitamin absorption and maintain the balance between cholesterol synthesis in the liver and cholesterol excretion. They are composed of a steroid structure and are primarily made in the liver by the oxidation of cholesterol. Cholesterol is also highly abundant in the human ovarian follicle, where it is used in the formation of the sex steroids. Methodology/Principal Findings: Here we describe for the first time evidence that all aspects of the bile acid synthesis pathway are present in the human ovarian follicle, including the enzymes in both the classical and alternative pathways, the nuclear receptors known to regulate the pathway, and the end product bile acids. Furthermore, we provide functional evidence that bile acids are produced by the human follicular granulosa cells in response to cholesterol presence in the culture media. Conclusions/Significance: These findings establish a novel pathway present in the human ovarian follicle that has the capacity to compete directly with sex steroid synthesis

Bile acid production in primary cultured human cumulus granulosa cells (CGC).

<p>Cholesterol entering granulosa cells may be used to form sex steroids or bile acids (Panel a). With increasing cholesterol concentration in media, CGC demonstrate dose-dependent bile acid content (Panel b).</p

Routine ketorolac at oocyte retrieval decreases postoperative narcotic use by more than 50%

Objective: To study the impact of routine ketorolac administration during oocyte retrieval on the proportion of patients who require postoperative narcotics for analgesia. Design: Retrospective cohort study. Setting: Single, university-affiliated infertility clinic. Patient(s): All women undergoing oocyte retrieval between July and November 2016 (non-ketorolac group [NKG]; n = 826) and April-August 2017 (ketorolac group, KG; n = 1780). Intervention(s): A single 30 mg intravenous dose of ketorolac was administered after the oocyte retrieval procedure. Main outcome measure(s): The number of patients who required postoperative narcotic analgesia, postoperative complication rate, and fresh embryo transfer pregnancy outcomes were examined. Result(s): In the KG, we found a significant decrease in the patients who required narcotics after oocyte retrieval compared with the NKG (12% KG vs. 25.5% NKG). We found no significant change in the clinical pregnancy rate (CPR) resulting from fresh embryo transfer after our intervention (NKG CPR 32.6%, KG CPR 32.4%). Furthermore, there was no increase in postoperative bleeding complications in the KG. Conclusion(s): Routine use of ketorolac at the time of oocyte retrieval may decrease the rate of postoperative opioid use without adversely impacting pregnancy and complication rates

Presence of enzymes and receptors regulating the classical and alternative pathways of bile acid synthesis.

<p>In the cytosol of granulosa cells, specific positive staining was registered for CYP7A1 (Panel A, image 1), CYP27A1 (Panel A, image 2), and CYP7B1 (Panel A, image 3). Specific positive staining for Inhibin β was used to confirm granulosa cell type (Panel A, image 4). In oocytes, specific positive staining was identified for CYP7A1 (Panel C, image 1), CYP7B1 (Panel C, image 2), and CYP8B1 (Panel C, image 3). The receptors known to regulate the bile acid synthesis pathway were also present in granulosa cells and oocytes. In granulosa cells, specific positive staining was registered for FXR (Panel B, image 1), LRH-1 (Panel B, image 2), and RXRα (Panel B, image 3). Using western blot of granulosa cells, a specific reacting band for the receptor LXRα was observed (Panel B, image 4). In human oocytes, specific positive staining was also demonstrated for the receptors FXR (Panel D, image 1), RXRα (Panel D, image 2), and LRH-1 (Panel D, image 3). Secondary antibody controls are presented in images 4 of Panels C and D.</p

Presence of receptors known to regulate the bile acids synthesis pathways.

<p>Protein presence was analyzed with receptor specific antibodies and either immunofluorescence (IF) or western blot (WB). (Present) – the receptor is present at a protein level; (absent) – the receptor is not present at a protein level.</p

Presence of bile acids in human ovarian follicular fluid.

<p>Bile acid concentration was measured in FF (Panel a), in FF compared to post-HCG serum (Panel b), and in pre-HCG serum compared to post-HCG serum (Panel c).</p