The Expression Of HGF As Hepatocytes Transplantation For Treatment Of Rat Acute Liver Failure

Objective :To master the way of isolating and purifying hepatocytes through an in-situ collagenase cycle perfusion . To investigate the function of Hepatocellular transplantation for treatment of D-galactosamine induced acute liver failure in rat . Methods : The mold of acute chemical liver failure: wistar rats were injected 10% D-galactosamine 1.6g/kg into abdominal cavity . Livers were perfusing with three different liquids , firstly D-Hanks including EDTA , secondly D-Hanks , finally collagenase type Ⅳ . Consistency of hepatocytes was 4×107 per milliliter . Some of hepatocytes were transplanted through spleens within two hours .The rats with acute liver failure induced by D- galactosamine were randomly divided into two groups . The groupⅠreceived 2×107 hepatocytes through spleens . the groupⅡreceived 0.5ml normal saline .Then HGF, liver function and histology of all rats were observed at 6h, 12h, 24h, 48h after hepatocytes transplanting respectively . Results: The hepatocytes isolated and purified yielded amount to 2.0-2.5×108 per rat . More than 95% of hepatoctyes identified with trypan blue were viable . The hepatocyte growth factor (HGF) of groupⅠis higher than that of the groupⅡwithin 24h (P<0.05),which of groupⅠcome to the peak at 12h . There were no difference between two groups at 48 h (P>0.05).The liver function of groupⅠ(ALT, TBil)is higher than that of the groupⅡwithin 12h (P<0.05) . The liver function of groupⅠ(ALT, TBil) gradually returned to the normal in 24 h .All of ALB determined in plasma , there were no difference between two groups(P>0.05) . Conclusions: The viability of hepatocytes isolated by the method mentioned above was high .The rat Allograft hepatocytes transplanted through spleens can stimulate liver of receivers to produce HGF , which makes its own hepatocytes regenerate within 24h after transplanting.