Figure 3

Treatment with bile acids resulted in mitochondrial dysfunction and trophoblast impairment.

(A–D) HTR8 cells were treated with 50 μM or 100 μM cholic acid (CA), deoxycholic acid (DCA) and chenodeoxycholic acid (CDCA) for 24 hours. (A) Cell viability, (B) ROS production, (C) mitochondrial membrane potential (ΔΨm) and (D) adenosine triphosphate (ATP) content were assayed using corresponding kits. The ATP level was shown as nmol/mg protein and the others were expressed as arbitrary units. (E,G) qRT-PCR assay was applied to detect mitochondrial gene transcripts levels (MT-CO1, MT-ND1 and MT-ND6 were taken as the representative of mitochondrial genes transcripts) in HTR8 cells treated with 100 μM CA, DCA, CDCA (E) and human placental tissues from normal pregnancies or intrahepatic cholestasis of pregnancy patients (G). The messenger RNA levels were calculated by normalisation to an internal control 18s rRNA. (F,H) Mitochondrial DNA copy number was quantified by quantitative polymerase chain reaction assay with normalisation to genomic DNA level in 100 μM CA, DCA and CDCA treated HTR8 cells (F) or human placentas (H). All experiments were performed in triplicate. Data were shown as mean ± standard error of mean. Statistical significance was determined by one-way analysis of variance followed by Dunnett’s post-hoc test, two-tailed Student’s t test or Mann-Whitney test. (*p < 0.05, **p < 0.01, ***p < 0.001 versus the Con or Normal group).