Allele association for <i>VDR</i> in patients with PSC and control subjects.

<p>Allele association for <i>VDR</i> in patients with PSC and control subjects.</p

Genotype counts for <i>VDR</i> polymorphisms (rs1544410, rs7975232, rs731236) in PSC patients and in controls.

<p>Genotype counts for <i>VDR</i> polymorphisms (rs1544410, rs7975232, rs731236) in PSC patients and in controls.</p

Demographic data of analysed subjects.

<p>Demographic data of analysed subjects.</p

Allelic analysis of rs1544410, rs7975232 and rs731236 in relation to SF-36, PBC-40 and PBC-27 domains.

<p>Allelic analysis of rs1544410, rs7975232 and rs731236 in relation to SF-36, PBC-40 and PBC-27 domains.</p

Relationship between the rs7975232 VDR polymorphisms and features of the SF-36, PBC-40 and PBC-27 questionnaires.

<p>Relationship between the rs7975232 VDR polymorphisms and features of the SF-36, PBC-40 and PBC-27 questionnaires.</p

Chenodeoxycholic (A), deoxycholic (B) and lithocholic acid-3glucuronide (C) conjugates are incorporated into HepG2 cells.

<p>HepG2 cells were cultured in the presence of vehicle (DMSO, control) or 100µM chenodeoxycholic (CDCA, <b>A</b>), deoxycholic (DCA, <b>B</b>), or lithocholic (LCA, <b>C</b>)- acid 3-glucuronides (G) for 1, 2, 4, 6, 8,12, 24 or 48H. The content in glucuronide conjugates into cell homogenates and culture media was resolved using LC-MS/MS.</p

Lithocholic (A,C,E&F), deoxycholic (C,E&F) and chenodeoxycholic (B,E&F) acids promote HepG2 cell death.

<div><p>HepG2 cells were exposed to vehicle (DMSO, control), lithocholic (LCA, <b>A, C, E & F</b>), chenodeoxycholic (CDCA, <b>B, E & F</b>), cholic (CA, <b>B</b>), deoxycholic (DCA, <b>C, E & F</b>), hyodeoxycholic (HDCA, <b>D</b>) or hyocholic (HCA, <b>D</b>) acids.</p> <p>(<b>A</b>) HepG2 cells were exposed to DMSO or 100µM LCA for up to 96H, and caspase-3 activity was assessed using the EnzChek Caspase-3 Assay Kit (Invitrogen).</p> <p>(<b>B</b>-<b>D</b>) HepG2 cells were cultured for 3H in the absence or presence of increasing concentrations (20 to 200µM) of CA (<b>B</b>), CDCA (<b>B</b>), LCA (<b>C</b>), DCA (<b>C</b>), HDCA (<b>D</b>) or HCA (<b>D</b>), and caspase-3 activity was assessed as described in the “Materials and Methods” section.</p> <p>(<b>E</b> and <b>F</b>) HepG2 cells were exposed to DMSO (Control), 100µM LCA, DCA or CDCA for 18 to 72H. Living, apoptotic and/or necrotic cells were then quantified through fluorescence-activated cell sorting (FACS) analyses using annexin V/propidium iodide-co-labeling, as represented on panel (<b>E</b>), and the relative abundance (expressed as percentage) of living (Live), apoptotic (Apop) and/or necrotic (Necr) cell populations were determined by dividing their quartiles by the total cell population (<b>F</b>).</p> <p>Statistically significant differences in vehicle <i>versus</i> treated cells were determined using the Student <i>t</i> test: *:p<0.05; </p><p>** p<0.01; *** p<0.001.</p><p></p></div

Chenodeoxycholic (A&D), lithocholic (B&E), deoxycholic (B&E) and hyodeoxycholic (C&F) acids reduce HepG2 cell viability.

<div><p>HepG2 cells were exposed to vehicle (DMSO, control) or increasing concentrations of chenodeoxycholic (CDCA, <b>A & D</b>), cholic (CA, <b>A & D</b>), lithocholic (LCA, <b>B & E</b>), deoxycholic (DCA, <b>B & E</b>), hyodeoxycholic (HDCA, <b>C & F</b>) or hyocholic (HCA, <b>C & F</b>) acids for 48 (<b>A</b>-<b>C</b>) or 96H (<b>D</b>-<b>F</b>).</p> <p>(<b>A</b>-<b>C</b>) Cell viability was determined using the MTS reduction assay.</p> <p>(<b>D</b>-<b>F</b>) The protein content was determined using the Pierce® BCA assay kit.</p> <p>Results, expressed relatively to vehicle-treated (DMSO, control) cells, represent the mean±S.D of 2 independent experiments performed in quadruplicate.</p> <p>Statistically significant differences in vehicle <i>versus</i> treated cells were determined using the Student <i>t</i> test: *:p<0.05; </p><p>** p<0.01; *** p<0.001.</p><p></p></div

Ether-glucuronide conjugates of lithocholic, deoxycholic and chenodeoxycholic acids are not cytotoxic.

<div><p>(<b>A</b> & <b>B</b>) HepG2 cells were exposed to vehicle (DMSO, control), or increasing doses (100, 150 and 200µM) of ether-glucuronides or unconjugated forms of lithocholic (LCA), deoxycholic (DCA), or chenodeoxycholic (CDCA) acids for 48 (<b>A</b>) or 3H. Cell viability (<b>A</b>) was determined using the MTS reduction assay and caspase-3 activity (<b>B</b>) was assessed as described in the “Materials and Methods” section.</p> <p>(<b>C</b> & <b>D</b>) HepG2 cells were exposed to vehicle (DMSO, control), or 200µM LCA, DCA, CDCA, LCA-3G, DCA-3G or CDCA-3G for 24 (LCA/LCA-3G) or 72H (DCA/DCA-3G and CDCA/CDCA-3G). Living, apoptotic and/or necrotic cells were then quantified through fluorescence-activated cell sorting analyses using annexin V/propidium iodide-co-labeling, as represented on panel (<b>C</b>), and the relative abundance (expressed as percentage) of living (Live), apoptotic (Apop) and/or necrotic (Necr) cell populations were determined by dividing their quartiles by the total cell population (<b>D</b>).</p> <p>Statistically significant differences in vehicle <i>versus</i> treated cells (*:p<0.05; </p><p>** p<0.01; *** p<0.001) or glucuronide- versus unconjugated-BA treated cells were determined using the Student <i>t</i> test: ¥ p<0.05; ^¥¥:p<0.01; ^¥¥¥:p<0.001.</p><p></p></div

Does transient elastography correlate with liver fibrosis in patients with PSC? Laennec score-based analysis of explanted livers

<p><b>Background and aims:</b> Previous studies demonstrated a close correlation between transient elastography (TE) and liver histology in chronic liver diseases. Data on the accuracy of TE in primary sclerosing cholangitis (PSC) remains scarce. Here, we investigated the association between TE, serum marker of liver injury and histology of explanted livers in PSC patients.</p> <p><b>Methods:</b> Thirty patients were prospectively recruited. TE (Fibroscan^®) and blood sampling were performed during evaluation for liver transplantation (LT); the second blood sampling was performed on the day of LT. Fibrosis of explanted livers according to the seven-point Laennec staging system and liver collagen contents were measured.</p> <p><b>Results:</b> TE correlated with Laennec stages of fibrosis (<i>p</i> = .001), collagen contents (<i>p</i> < .001) and with diameter of thickest septa (<i>p</i> = .034) in explanted livers. It also correlated with serum indices of liver injury, namely AST, bilirubin as well as FIB-4 and APRI scores (all <i>p</i> < .05). In a multivariate model, only liver fibrosis, according to either Laennec score (<i>p</i> = .035) or collagen contents (<i>p</i> = .005), was significantly associated with TE. Finally, patients with cirrhosis had increased liver stiffness (<i>p</i> = .002) and the TE cut-off of 13.7 kPa showed the best predictive value (AUC = .90, 95% CI: 0.80–1.00, <i>p</i> < .001) for detecting cirrhosis.</p> <p><b>Conclusions:</b> TE correlates with liver fibrosis and markers of liver injury in patients with PSC. However, liver fibrosis seems to be the strongest predictor of liver stiffness assessed with TE. Hence, we postulate that TE is a reliable tool for non-invasive monitoring of PSC.</p