The effect of bile salts and calcium on isolated rat liver mitochondria.

Intact mitochondria were incubated with and without calcium in solutions of chenodeoxycholate, ursodeoxycholate, or their conjugates. Glutamate dehydrogenase, protein and phospholipid release were measured. Alterations in membrane and organelle structure were investigated by electron paramagnetic resonance spectroscopy. Chenodeoxycholate enhanced enzyme liberation, solubilized protein and phospholipid, and increased protein spin label mobility and the polarity of the hydrophobic membrane interior, whereas ursodeoxycholate and its conjugates did not damage mitochondria. Preincubation with ursodeoxycholate or its conjugate tauroursodeoxycholate for 20 min partially prevented damage by chenodeoxycholate. Extended preincubation even with 1 mM ursodeoxycholate could no longer prevent structural damage. Calcium (from 0.01 mM upward) augmented the damaging effect of chenodeoxycholate (0.15-0.5 mM). The combined action of 0.01 mM calcium and 0.15 mM chenodeoxycholate was reversed by ursodeoxycholate only, not by its conjugates tauroursodeoxycholate and glycoursodeoxycholate. In conclusion, ursodeoxycholate partially prevents chenodeoxycholate-induced glutamate dehydrogenase release from liver cell mitochondria by membrane stabilization. This holds for shorter times and at concentrations below 0.5 mM only, indicating that the different constitution of protein-rich mitochondrial membranes does not allow optimal stabilization such as has been seen in phospholipid- and cholesterol-rich hepatocyte cell membranes, investigated previously.

TUDCA and UDCA are incorporated into hepatocyte membranes: different sites, but similar effects.

The aim of this paper is to point out that: 1) CDCA and DCA increase the polarity of cell membranes and cause the release of cholesterol and phospholipid from the membranes; 2) the extent of this damage is inversely correlated with the cholesterol content of the membrane investigated; 3) UDCA, TUDCA and GUDCA decrease membrane polarity; 4) they prevent membrane damage when added prior to CDCA or DCA; 5) UDCA appears to be incorporated into the apolar domain of the membrane, TUDCA, GUDCA into the interface; 6) UDCA decreases HLA class I expression on hepatocyte membranes; 7) CDCA induces GLDH-release from liver mitochondria and increases mitochondrial membrane polarity and mobility; and 8) UDCA reduces the release of GLDH from mitochondria caused by CDCA.