Expression of the MT1 melatonin receptor subtype in human coronary arteries.

Previous experimental data suggest a possible influence of melatonin on the circulatory system of animals after binding to G-protein coupled melatonin receptors. The present study sought to investigate whether the melatonin receptor, mt1, is expressed in human coronary arteries derived from healthy heart donors (n = 8). Expression of the mt1-receptor was studied in sections of isolated coronary arteries by a reverse transcriptase-polymerase chain reaction (RT-PCR) and Western immunoblot technique. The analyses of the results from both methods indicated the presence of the mt1-receptor in all of the subjects. Referring to these data we assume that melatonin regulates physiological processes in human coronary arteries after receptor binding.

Influence of hypotonic stress on the biliary excretion of doxorubicin in Wistar and TR- rats.

In tumor cells, doxorubicin (DOX) is excreted by P-glycoprotein (P-gp) and the multidrug resistance-associated protein (mrp). Both transporters might also be involved in cellular regulatory volume decrease (RVD). To study the hepatobiliary excretion of DOX during RVD, isolated livers of Wistar and multidrug resistance-associated protein 2 (mrp2)-deficient TR- rats were first perfused with an isotonic and later with a hypotonic medium. In both rat strains, DOX is effectively excreted into the bile. Within 30 minutes, the biliary excretion of DOX-derived fluorescence steadily increased to 1.1+/-0.11 and 0.84+/-0.05 nmoles/min . g liver in Wistar and TR- rats, respectively. Under hypotonic conditions, DOX excretion followed the biphasic-increase in bile flow. Excretion was increased to 136+/-19% and 176+/-9% (first peak) and to 141+/-11% and 157+/-14% (second peak) in Wistar and TR- rats, respectively. Our data show that in the liver of both strains, exposure to a hypotonic medium stimulates DOX excretion, presumably by activation of P-gp and mrp2 during RVD.

Characterization of autoantibodies against uridine-diphosphate glucuronosyltransferase in patients with inflammatory liver diseases.

Uridine diphosphate glucuronosyltransferase (UGT) was identified as an antigenic target in a subgroup of liver-kidney microsomal autoantibodies and was termed LKM-3. To evaluate the nature of LKM-3 antibodies, we screened sera from 80 untreated patients with autoimmune hepatitis (AIH) type 1 and 2, primary biliary cirrhosis (PBC), AIH/PBC, hepatitis C virus (HCV) infection, and 12 healthy individuals (controls) against 7 recombinant human UGT isoenzymes (UGT1A1, UGT1A4, UGT1A6, UGT1A7, UGT1A9, UGT1A10, and UGT2B7). Autoantibodies reacting against various UGT isoenzymes were observed in sera from 3 of 18 AIH type 2 and 1 of 27 of the HCV patients. The anti-UGT-positive sera from AIH type 2 patients revealed the strongest immunoreactivity against UGT1A1, the main UGT-isoform involved in the bilirubin glucuronidation. Additionally, these sera were able to block UGT-mediated substrate glucuronidation in vitro. The prevalence for UGT1A1 was shown by 2 independent techniques: (1) UGT1A1 was identified as the main antigen by Western blotting. Preabsorption of sera with UGT1A1 prevented reaction against all tested UGT-isoforms. (2) In vitro immunoinhibition experiments showed that glucuronidation of the anticancer drug flavopiridol by UGT1A1 was more strongly inhibited than its UGT1A9-mediated biotransformation. In contrast, the serum from the HCV-patient reacted predominately with UGT1A6, and moreover, the immunoreactivity pattern was different from that of the AIH group. To summarize, we show the subtype preference of antibodies against UGT1A1 in a subgroup of AIH type 2 patients. These autoantibodies inhibit UGT-mediated glucuronidation in vitro, but it is unlikely that anti-UGT antibodies will have a marked effect on the patients capacity for drug biotransformation, as serum bilirubin levels in patients remained within the normal range.

24h variation in the expression of the mt1 melatonin receptor subtype in coronary arteries derived from patients with coronary heart disease.

Previous studies presented evidence for impaired nocturnal secretion and synthesis of melatonin in patients with coronary heart disease (CHD). This study aimed to investigate whether the melatonin receptor subtype mt1 is differentially expressed in coronary arteries derived from patients with CHD (n = 9) compared to patients with dilative cardiomyopathy (CMP; n = 10) who served as controls. Expression of the mt1 receptor was studied in sections of isolated coronary arteries by a reverse transcriptase-polymerase chain reaction (RT-PCR) and a Western immunoblot technique. In addition, the data from the Western blotting of 15 patients were interpolated against the exact time of aortic clamp to study the 24h expression of the mt1 receptor. The analyses of the results from both methods indicated the presence of the mt1 receptor in all of the individuals. No statistically significant difference was observed in the receptor expression between patients with CHD and those with CMP (in arbitrary units: 3.39 +/- 3.08 versus 3.91 +/- 2.78). Expression of the melatonin receptor in the coronary arteries of the whole patient group presented a 24h variation, with the lowest values detectable after 02:00 up to the late morning hours and a progressive increase beginning after 13:00 until 00:00 (mesor = 3.66, amplitude = 3.23, acrophase = 20.45, P = .0003). When studying the 24h variation in patients with CHD and CMP separately, a nearly similar circadian course was observed. In conclusion, we demonstrated for the first time a 24h variation of a melatonin receptor subtype in human vessels. Furthermore, in relation to our results, we suggest that the expression of the mt1 melatonin receptor in the coronary arteries is probably not impaired in patients with CHD.