805 Activation, proliferation and differentiation of progenitor cells in the rat model with D-galactosamine and biliary duct ligation

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AASLD ABSTRACTS

HEPATOLOGY, October 2003

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ACTIVATION, PROLIFERATION AND DIFFERENTIATION OF PROGENITOR CELLS IN THE RAT MODEL WITH DGALACTOSAMINE AND BILIARY DUCT LIGATION. Qing

THE EFFECT OF RECOMBINANT FACTOR VIIA AND FRESH FROZEN PLASMA ON THE INR IN PATIENTS WITH ACUTE AND CHRONIC LIVER FAILURE. David J Quan, Nathan M

Xie, Yumin Xu, Chungen Yah, Huijuan Zhou, Jiacheng Xiao, Xiaqiu Zhou, Hong Yu, (~'ng Guo, Shanghai Ruijin Hospital, Shanghai Second Medical University, Shanghai, China Objective: A SD rat model with acute hepatic injury is established with D-galactosamine(D-GalN) or D-GaIN combined with common biliary duct ligation(CBDL) to identify the activation, proliferation and differentiation of hepatic stem ce11. Methods: The Male SD rats were treated with various doses of D-GaIN intraperitoneally(ip) or D-GaIN combined with CBDL, and A single injection of 5'-bromodeoxyuridine(Brdu) was given ip at the dose of 50mg/kg body weight to the animals I hour before sacrifice. Blood were drawned for liver function analysis. Morphological evaluation of the cells was performed with HE staining and electron microscope. Different expression of hepatic stem cells in different time points and groups was detected by RT-PCR or immunohistochemistry. Results: (1)The liver functions of a11 the rats in different groups and times(from day 1 to day 6) were abnorma1(P<0.05).The ALT value of the group'D-GaIN2.0g/kg body weight' is the highest among all the groups(P<0.01),and the ALT value on day 2 is the highest among a11 the days(P<0.01).The TB(total bilirubin) value of the group'D-GaIN2.0g/kg body weight plus CBDL' is the highest among a11 the groups(P<0.01), and the TB value on day 2 is the highest among a11 the days(P<0.01). (2) No hepatic oval cells(HOC) were observed in controls with HE staining. Focal hepatocyte degeneration and necrosis was conspicuous in a11 the groups and days, and the hepatocyte plates were disorganized. Proliferation of HOC began in the periportal spaces on day two and gradually invaded into the 1obule. Proliferating HOC formed rows, clusters, and duct-like structures. By day 5, the liver began to restore its normal lobular structure. The result of electron microscope showed that the HOC was small with scant cytoplasm and oval shaped, pale, and homogeneously stained nuclei.(3)With immunohistochemistry, HOC was characterized with brown staining in the cytoplasm for cytokeratin 7,8,19,AFP, ALB, Hep,OV-6 separately. BrdU-labeled nuclei of proliferating cells were demonstrated in the rat following induction of liver damage.Cells were counted if they fufiled the morphological criteria for oval cells and showed cytoplasmic staining of OV-6. The number of HOC was the largest in group'D-GaIN2.0g/kg body weight +CBDL' among a11 the groups(P<0.01), and the number on day 3 was the highest among a11 the days(P<0.05).(4)The result of RT-PCR showed the expression of GGTmRNA was much more than the controls, especially in the group'D-GaIN1.4g/kg body weight plus CBDL'and the group'D-GaIN2.0g/kg body weight plus CBDL'(P<0.05), and it was the highest on day 3 among a11 the days(P<0.05). Conclusions: The induced model of HOC activation can be established with D-GaIN or D-GaIN combined with CBDL, such as D-GaIN1.4g/kg body weight, D-GaIN2.0g/kg, D-GaIN1.4g/kg plus CBDL, D-GalN2.0g/kg plus CBDL. The activation of HOC is more obvious especially following D-GalN2.0g/kg plus CBDL. Treatment. Morphological and molecular biological analysis revealed that there are hepatic stem cells and they can differentiate into hepatocytes and bile duct cells with the liver injury. These findings will contribute to the further clinical application of the hepatic stem cells. Disclosures: Qing Guo - No relationships to disclose Jiacheng Xiao - No relationships to disclose Qing Xie - No relationships to disclose Yumin Xu - No relationships to disclose Chungen Yah - No relationships to disclose Hong Yu - No relationships to disclose Huijuan Zhou - No relationships to disclose Xiaqiu Zhou - No relationships to disclose

Bass, Ryutaro Hirose, University of California San Francisco, San Francisco, CA Recombinant factor VIIa (rVIIa, Novoseven) is being more frequently used for the correction of severe coagulopathy in patients with liver disease, particularly to enable invasive procedures and control bleeding without the fluid volume load associated with fresh frozen plasma (FFP). However, the clinical value, cost effectiveness and appropriate dosage of rVIIa in patients with liver disease remains unestablished. The purpose of this study was to evaluate the dose-response relationship of rVIIa, with and without FFP on the international normalized ratio (INR) in patients with coagulopathy from acute and chronic liver disease. METHODS: Medical records for patients admitted to the Liver Transplant Service at UCSF Medical Center who received factor rVIIa between 9/2001 to 5/2003 were reviewed. Patients who received factor rVIIa intra-operatively for hemostasis were excluded. Patients were divided into two groups (NO FFP, WITH FFP), then stratified into three dose range groups (<40mcg/kg, 40-80mcg/kg, and >80mcg/kg per dose). Comparisons were made for the MELD score, pre- and post-treatment INR, using paired and unpaired Student's t-tests with significance at p < 0.05. RESULTS: Thirty-six patients received 71 doses of rVIIa. Response to all doses was included in the analysis. Indications for rVIIa dosing were periprocedural (83%), bleeding (10%), elevated INR (4%), and other (2%). Baseline INR and MELD scores were similar among all groups. The results are shown in the Table ( N - d o s e s , M e a n ± SD). Significant reduction in INR was observed only in the <40mcg/kg groups (±FFP) and the 40-80mcg/kg/group who received FFP. The addition of rVIIa to FFP resulted in a significant reduction in the INR value only in the 40-80mcg/kg/dose group. CONCLUSIONS: Administration of rVlIa results in a reduction in INR in patients with coagulopathy of liver disease. The addition of rVIIa to FFP may result in a greater reduction in INR than w h e n given alone. The magnitude of reduction in the INR value was similar among all dose groups, and significant only in the lower dosage ranges. Reduction in INR in patients with liver disease may be achieved with rVIIa most effectively at relatively low doses particularly w h e n combined with FFP. Eff~ ~f ee~a

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p<0.05: *Pre v s Post INR; § N O FFP vs W I T H FFP

Disclosures: Nathan M Bass - No relationships to disclose Ryutaro Hirose - Novo Nordisk: Consultant/Advisor David J Q u a n - No relationships to disclose