Prophylactic Intra-Uterine β-Cyclodextrin Administration during Intra-Uterine Ureaplasma parvum Infection Partly Prevents Liver Inflammation without Interfering with the Enterohepatic Circulation of the Fetal Sheep

Abstract

Chorioamnionitis can lead to inflammation and injury of the liver and gut, thereby predisposing patients to adverse outcomes such as necrotizing enterocolitis (NEC). In addition, intestinal bile acids (BAs) accumulation is causally linked to NEC development. Plant sterols are a promising intervention to prevent NEC development, considering their anti-inflammatory properties in the liver. Therefore, we investigated whether an intra-amniotic (IA) Ureaplasma parvum (UP) infection affected the liver and enterohepatic circulation (EHC) and evaluated whether an IA administered plant sterol mixture dissolved in β-cyclodextrin exerted prophylactic effects. An ovine chorioamnionitis model was used in which liver inflammation and the EHC were assessed following IA UP exposure in the presence or absence of IA prophylactic plant sterols (a mixture of β-sitosterol and campesterol dissolved in β-cyclodextrin (carrier)) or carrier alone. IA UP exposure caused an inflammatory reaction in the liver, histologically seen as clustered and conflated hepatic erythropoiesis in the parenchyma, which was partially prevented by IA administration of sterol + β-cyclodextrin, or β-cyclodextrin alone. In addition, IA administration of β-cyclodextrin prior to UP caused changes in the expression of several hepatic BAs transporters, without causing alterations in other aspects of the EHC. Thereby, the addition of plant sterols to the carrier β-cyclodextrin did not have additional effects.

The Effect of MSTN Mutation on Bile Acid Metabolism and Lipid Metabolism in Cattle

“…In cattle, the primary bile acids produced are cholic acid and chenodeoxycholic acid [ 61 ], whereas other species, such as mice and humans, produce muricholic acid in addition to these two primary bile acids [ 62 ]. Moreover, the enterohepatic circulation of bile acids is less extensive in ruminants than in monogastric animals [ 42 , 63 ]. These differences must be taken into account when investigating the influence of MSTN mutation on bile acid metabolism in cattle.…”

Chorioamnionitis induces hepatic inflammation and time-dependent changes of the enterohepatic circulation in the ovine fetus

“…A Hematoxylin and Eosin (H&E) staining was performed which an independent pathologist blinded to the experimental set-up analyzed as previously described 25 . In short, the slides were qualitatively scored pathologically on a 0 to 4 scale for hepatic sinusoidal dilatation, shape and size of central veins and number and location of extramedullary hematopoietic clusters.…”

“…RNA extraction and quantitative real-time polymerase chain reaction (qPCR) was performed as previously described 25 . In short, RNA was extracted from snap frozen liver and terminal ileal tissue using TRI reagent (Thermo Fisher Scientific)/chloroform extraction.…”

“…Total bile acid assay. As previously described 25 , total bile acids (tBAs) were measured in plasma, liver homogenate and terminal ileum homogenate by an enzymatic cycling method, according to the manufacture protocol (Total Bile Acids Assay kit, Diazyme Laboratories, Poway, CA, USA). tBAs in liver and ileum homogenate were corrected for protein content, which were measured with a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA).…”