ABSTRACT
AIM: To activate hepatic stellate cells (HSC) in vitro in precision-cut liver slices (PCLS) stimulated by acetaldehyde for studying and screening anti-fibrotic drugs. METHODS: PCLS were prepared by the vibratome, and incubated with 700 ?m?L~-1 acetaldehyde for 0, 2, 4 and 6 h. The medium and homogenate were retained to determine glutathione S-transferase (GST) activity, lactate dehydrogenase (LDH) leakage and hydroxyproline (Hyp) content. PCLS were prepared for paraffin sections. The expression of ?-smooth muscle actin (?-SMA) was evaluated by immunohistochemistry, and the result was analyzed with image analysis software. RESULTS: The leakages of GST and LDH were increased significantly compared with those in 0 h group (P
ABSTRACT
AIM: To establish the technique of precision-cut fibrotic liver slice (PCLS) and grope the optimal cultural conditions for researching the liver xenobiotic metabolism in vitro and the drug interaction. METHODS: Complex factors (higher fat diet, alcohol and CCl 4) were used to make the animal model of liver fibrosis. Fibrotic liver slices were prepared and cultivation system was established. Lactate dehydrogenase (LDH) leakage, glutathione S-transferase (GST) activity and 3[4,5-Dimethythiazole-2-yl]-2,5- diphenyltetrazolium bromide (MTT) reduction were chosen as indexes to assess the viability of the slice in different thickness, medium pH and cultural time. RESULTS: Rats were in earlier hepatic fibrosis after administration for 3 weeks. When the thickness of slices was 300 ?m and medium pH was 7.0, the LDH leakage, GST activity and MTT reduction could maintain on a steady level in 6 h. CONCLUSION: A 300 ?m of thickness, 7.0 of medium pH and 6 h of cultural time are the optimal slicing and culturing conditions for fibrotic liver slice.