The C-Terminal Peptide Of Anaphylatoxin C3a Enhances Liver Cell Proliferation In A Rat Partial Liver Transplantation Model

Objective: To establish the animal model of partial orthotopic liver transplantation in rats, and analyze the variability of liver regeneration in different groups. After the rat model of 60% partial orthotopic liver transplantation have been established, nonapeptide, based on homologies to the C terminus of C3a,was administered intraperitoneally. The serum levels of alanine aminotransferase(ALT),proliferating cell nuclear antigen labeling index (PCNA LI) and morphological change in liver sections were detected in order to evaluate the effects of C3a nonapeptide on hepatic regeneration. Further more, the mRNA levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) were detected by real-time PCR so as to explore the mechanism of C3a nonapeptide in hepatic regeneration.Methods: 1. Establishment of the rat model: 60% partial orthotopic liver transplantation model with left lobe and double caudal lobe resected was established by removing full liver and implanting into a 60% partial liver with the modified two-cuffed vascular anastomoses technique of Kamada. 2. Sprague-Dawley (SD) to SD Rats model of 60% partial orthotopic liver transplantation were divided into two groups randomly. In group A, 100ug C3a nonapeptide was administered intraperitoneally every 6 hours after liver transplantation;And in group B, same dose of physiological saline for injection was administrated as a control. 3. 6 rats were sacrificed randomly at the time of 12h,24h,48h and 72h respectively after operation in both groups. The serum levels of ALT was detected by an automatic biochemistry analyzer, the pathohistological change of grafts were observed by light microscope , the expression of PCNA was detected in liver sections by immunohistochemistry assay, and the expression of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) mRNA was detected by real-time RT-PCR technique.Results: 1. Stable rat model of partial orthotopicd liver trans- plantation had been established successfully. There was no significant difference between groups A and B in the duration of operation, cold-ischemia time, anhepatic phase time and loss of blood. 2. Serum ALT level of group A was significantly lower than that of group B at 24h and 48h after operation. 3. The peak of hepatocyte PCNA LI in both groups was both at 48h after operation, the level of PCNA LI of group A was significantly higher than that of group B at 24h and 48h after operation, and no differences were observed at other time phases between group A and group B. 4. The expression peak of TNF-αand IL-6 mRNA in hepatic tissue in both groups was at 48h after operation, the level of TNF-αand IL-6 mRNA of group A was significantly higher than that of group B at 24h,48h and 72h after operation.Conclusions: 1,C3a nonapeptide can promote the regeneration of hepatocyte at the early stage of partial orthotopic liver transplantation, decrease the level of serum ALT and preserve hepatic function. 2,The mechanism of C3a nonapeptide in promoting hepatocyte regeneration could probably be to up-regulate the expression level of TNF-αand IL-6 mRNA in the early stage of partial orthotopic liver transplantation in hepatic tissue. This may contribute to explorations on full liver size and function recovery after Liver donor liver transplantation (LDLT).