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181.
While intrahepatic bile duct epithelial cells secrete bile through transport of ions and water, the physiological mechanisms regulating ductular bile secretion are obscure, in part because of the lack of suitable experimental models. We report here the successful micropuncture of the lumen of isolated intrahepatic bile ducts and direct measurements of ductular ion secretion. Intact, polarized bile duct units (BDUs) were isolated from livers of normal rats by enzymatic digestion and microdissection. BDUs were cultured and mounted on a microscope in bicarbonate-containing buffer, and the lumens were microinjected with 2',7'-bis(2-carboxyethyl)-5-(and -6)carboxyfluorescein (BCECF)-dextran. Lumenal pH was measured by ratio imaging of BCECF fluorescence using digitized video fluorescent microscopy. After 36 hr in culture, the ends of BDUs sealed, forming closed compartments. After lumenal microinjection of BCECF-dextran, fluorescence was stable at the pH-insensitive wavelength, indicating no dye leakage. Serial changes in pH of extralumenal buffers containing pH-gradient collapsing ionophores allowed us to establish reliable standard curves relating fluorescence ratio to lumenal pH (r = 0.99; P < 0.001). By this approach, the basal pH inside the lumen of BDUs was 7.87 +/- 0.08 units (n = 9), 0.47 unit higher (P < 0.001) than the bathing buffer pH. Addition of 100 microM forskolin increased (P = 0.02) the lumenal pH from 7.78 +/- 0.06 to 7.97 +/- 0.06 units (n = 5); the forskolin effect was completely abolished by incubation of BDUs in HCO3-/CO2-free buffer. Moreover, forskolin caused a 50-fold increase in cAMP levels in BDUs. The observations are consistent with cAMP-dependent, active lumenal HCO3- secretion by BDUs. Furthermore, they demonstrate the suitability of the BDU model for studying regulatory and mechanistic aspects of ductular bile secretion. 相似文献
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Tanshinone II-A (TSII-A) isolated from the root of Salvia miltorrhiza Bunge, a traditional medicine in China, is a derivative of phenanthrenequinone, which is known to have antioxidant properties. In the present study, effects of TSII-A on DNA damage by lipid peroxidation were investigated using liver cells, labeled with [3H] arachidonic acid, in the presence of FeCl2-DTPA. The results show that the nuclear DNA isolated from treated cells had higher radioactivity compared to controls and the radioactivity increased with longer incubation times. Purified lipid-DNA adducts had a characteristic fluorescent spectra and showed a decrease of hyperchromicity and melting point. TSII-A could inhibit the association of peroxidation products with DNA in liver cells and prevent a decrease in cell viability and in the the activity of O6-methylguanine acceptor protein with increasing incubation time. Compared with other antioxidants, TSII-A had a higher inhibitory ratio, which was similar to vitamin E and butylated hydroxy-toluene (BHT), but markedly stronger than NaN3, mannatol, and superoxide dismutase (SOD). These data suggest that TSII-A represents a new and effective antioxidant that inhibits the association of lipid peroxidation products with DNA. Its protective effect may be through breaking the chain reactions of peroxidation by scavenging lipid free radicals, thereby decreasing their cytotoxicity. 相似文献
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K Brasso S Friis SK Kjaer T J?rgensen P Iversen 《Canadian Metallurgical Quarterly》1998,51(4):590-594
OBJECTIVES: To review the trends in prostate cancer (PC) incidence and mortality rates in Denmark during a 50-year period. METHODS: A population-based register study was performed of all new cases of PC recorded in the Danish Cancer Registry from 1943 to 1992. RESULTS: The age-standardized incidence rate for PC increased from 11.5/100,000 in 1943 to 1947 to 30.9/100,000 in 1988 to 1992. Age-specific incidence rates increased in all age groups over 50 years of age. Mortality rates increased from 13.5/100,000 in 1953 to 1957 to 17.8/100,000 in 1988 to 1992. No major changes in the distribution of age, stage at the time of diagnosis, or in diagnostic procedures were found, indicating that the observed change in incidence rates was not caused by attempted early detection or changes in diagnostic strategy. CONCLUSIONS: Our data suggest that the increased PC incidence observed during the period of cancer registration in Denmark represents a true increase in the number of patients with clinical PC. 相似文献
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It has been demonstrated that exposure to mercury or cadmium compounds causes alterations in the glutathione system in a model glial cell line, C6. Here we report that two organic tin compounds, triethyltin (TET) and trimethyltin (TMT), are also toxic to these cells with EC50 values for cell death of c. 0.02 microM and 0.8 microM respectively. Exposure for 24 h to either of these compounds at sub-toxic concentrations caused increases in the amount of reduced glutathione (GSH) per cell. Increases in glutathione-S-transferase enzyme activity were also demonstrated after TET or TMT exposure. This suggests that glutathione increases occur in glial cells after toxic insults below that required to cause cell death, possibly acting as a protective mechanism. To test whether GSH plays a role in organotin-induced cell death we manipulated GSH in the culture media or via intracellular GSH and looked at the effects on sensitivity to TET or TMT toxicity. Adding GSH to the culture media did not protect the cells. Depletion of intracellular GSH with buthionine-[S,R] sulphoximine did not alter cytotoxicity of TET or TMT. However, pre-treatment with (-)-2-oxo-4-thiazolidine carboxylic acid (OTC), which increases intracellular GSH levels, protected the cells against both compounds. The EC50 for TMT was increased from 0.77 to 1.8 microM, a 2.3-fold shift, whereas the EC50 for TET was increased > 20-fold, from 0.022 to 0.47 microM. One interpretation of these results is that GSH protects cells against the toxicity of organic tin compounds without reacting directly with them to any significant extent. Under conditions where GSH is depleted, additional protective mechanisms may be active. 相似文献
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