| [Objective] To study the different expression of the HGF/c-met pathway and its related factors during liver regeneration after partial hepatectomy(PH) in cirrhotic rats and explore the reasons for the slow liver regeneration after partial hepatectomy in cirrhotic rats.[Methods]Part1:To establish the model of cirrhotic rats after partial hepatectomy with independent liver lobe congestion:we use carbon tetrachloride to induce liver cirrhosis in rats firstly, then perform70%liver resection in cirrhotic rats.Part2:To investigate the different expression of the HGF/c-met pathway and its related factors during liver regeneration after partial hepatectomy in cirrhotic rats.60healthy aldut male Wistar rats were divided into two groups:Group N were merely performed with70%PH; Group C were performed with70%PH after induced liver cirrhosis.Rats in each group after3hours,6hours,12hours,24hours and48hours were randomly sacrificed5. Blood samples and liver samples were obtained when rats were sacrificed after anesthesia and cryopreserved-80℃. The remaining part of liver was fixed by formaldehyde, make conventional HE staining and immunehistochemical staining (C-met). Get serum after standing and centrifugalization, then use it to detect hepatocyte growth factor (HGF) and hepatocyte growth factor activator (HGFA) by enzyme-linked immunosorbent assay (ELISA). Detect the HGF mRNA,C-met mRNA, HAI-1mRNA and HAI-2mRNA by real-time reverse transcription polymerase chain reaction (RT-PCR). The results of two groups were analyzed using SPSS19.0statistical analysis software, observe compa rison of various indicators of group differences in liver regeneration (independent samples t test).[Results]Part1:The model of70%partial hepatectomy in cirrhotic rats was stable;Part2:1. The serum HGF and liver HGF mRNA were significantly higher in group C than in group N (P<0.05). HGF began to increase after3h, and reached the peak at12h in group N.HGF began to increase after12h in group C, and reached the peak at24h. With the passage of time, two groups of HGF all increased firstly and then decreased.2.The serum level of HGFA were significantly lower in group C than in group N (P<0.05). The two groups of rats serum HGFA dropped to the lowest level after12h, then gradually picked up, to the48h returned to normal.3.The C-met protein and C-met mRNA were significantly lower in group C at any time point (P<0.05). C-met started to decline after6h in group N, fell to the lowest at24h, and began to rise after24h. With the passage of time,C-met started to decrease after12h.4. The expression of HAI-1mRNA were significantly higher in group C at any time point after PH (P<0.05). The expression of two groups started to increase at6h after operation, and reached the peak at12h, then gradually returned to normal level.5.There was no significant difference of the expression of HAI-2mRNA between the two groups at any time point after PH (P<0.05), the arbitrary two time points in one group were also not statistically different (P<0.05).[Conclusions]1.This investigation choose a simple and steady method to induce liver cirrhosis rat models through a large number of pre experiment, then perform the70%partial hepatectomy in cirrhotic rats. This animal model is an ideal model to investigate the regulation of liver regeneration after hepatectomy in cirrhotic rats with strong operability and high achievement ratio.2. Compared to the control group, the expression ability of the HGF/C-met pathway in cirrhosis rats is lower than normal rats, which may result in the slow liver regeneration after partial hepatectomy.3.The reducing of the synthesis of HGF is regulated by HGFA and HAI-1in cirrhotic rats, there may be a feedback effect between the three.But the reason of the sustained high expression of HAI-1in liver cirrhosis is unknown,which needs further study.4. HAI-2may not participate in the injury and repair of liver tissue. |
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