| Partial hepatectomy is a classic therapeutic method for liver transplantation and liver cancer treatment.The powerful regenerative function of liver is the guarantee for the recovery of liver function after partial hepatectomy.Laparoscopic surgery is,more and more widely,applied,because of its advantages which are minimally invasive,clear view of operation and little stimimulation.Up to now,the laparoscopic technology has made great progress,and more and more attention has been paid to the es Tab.lishment of animal models by laparoscopy.Presently,our has successfully bulid miniature pig gastrointestinal fistula animal model,miniature pig renal failure model and minniature pig liver injury model by utilizing laparoscopic technology,which provided a large number of scientific basis for clinical research and comparative medicine.This experiment is based on minniature pig laparoscopic liver injury model,injecting HRS through the portal vein,through the detection of relevant indicators,to evaluate of influence of HRS on liver regeneration after liver injury.24 miniature pigs were randomly divided into three groups,the sham operation group(group A),model group(group B),HRS intervention group(group C).Group A only esTab.lished pneumoperitoneum and surgical pathway,turning the hepatic lobe,no other operation,sutured incision of abdominal wall.Group B took laparoscopic right hepatic ischemia 60 minutes with left hemihepatectomy.Group C was injected HRS through portal vein at 10 minutes before reperfusion,and the other operations were same with Group B.The experiment through es Tab.lishing pneumoperitoneum,selecting the operation path,separating the cystic duct and artery,taking 60 minutes right hepatic ischemia and completing portal vein intubation,and at 10 minutes before reperfusion injecting HRS(10ml/kg).After 60 minutes ischemia taking laparoscopic left hemihepatectomy,subsequently,peritoneal lavage,Abdominal suture and extraction of the liver parenchyma should be taken.HRS(10ml/kg)will continue to inject at postoperative 1d,2d,3d.Venous blood was collected from group A,group B,group C,seperately,at preoperative,postoperative 1d,2d,3d,5d,7d.Liver tissue was collected from the groups seperately at preoperative and postoperative 1d,3d,7d.Blood samples was used to detected liver function(ALP,LDH,TP),hyaluronic acid,and vascular growth factors(VEGF,Ang-1,Ang-2).Liver tissue was used for histological observation with HE staining,immunohistochemistry fordetecting PCNA and detecting the changes of liver regeneration related genes(HGF,Cyclin-D1,TGF-?1)expression with q RT-PCR.The results are as follows:All laparoscopic operation were successfully completed,and the postoperative recovery was good.Compared with preoperative values,the serumconcentration of ALP,LDH,TP after operation are extremely significant difference(P<0.01)atpostoperative 1d,2d,the concentration of ALP,LDH in group B is higher than group after operation,the concentration of ALP at postoperative 1d,2d is significant difference(0.01<P<0.05),and the LDH at postoperative1 d is extremely significant difference(P<0.01),which shows that HRS is benefit for liver function recovery.Compared with preoperative values,the concentration of HA in serum increased firstly and then decreased after operation and reached the maximum atpostoperative 1d which is extremely significant difference(P<0.01),the concentration of HA in group B is higher than group C after operation which is extremely significant difference(P<0.01)at postoperative 1d,2d,3d,that shows HRS can alleviate the injury of LESC.After the injury,compared with preoperative,hepatocyte appears apoptosis,necrosis,inflammatory infiltration is most serious,at postoperative7 d is gradual recovery,the organization structure is more complete and closed tonormal tissue in group C at postoperative 7d,while in group B,there are still more inflammatory cells infiltration,which illustrates HRS is benefit for the recovery of liver structure.Compared with preoperative values,the concentration of VEGF,Ang-1,Ang-2 in serum increased firstly and then decreased after operation and group B is lower than group C,the concentration of VEGF,Ang-1 is extremely significant difference(P<0.01)at postoperative 3d,the Ang-2 is extremely significant difference(P<0.01)at postoperative 3d and 5d,that shows the up-regulation of VEGF,Ang-1,Ang-2 may be caused by HRS.Compared with preoperative,the expression of PCNA in Hepatocytes reachedmaximum at postoperative 1d and is extremely significant difference(P<0.01)which group B is lower than group C,also,which is significant difference(0.01<P<0.05)at postoperative3 d,which shows HRS can promote the expression of PCNA.Compared with preoperative,the expression of HGF and Cyclin-D1 after operation in group B is lower than group C,reached maximum at postoperative 1d(P<0.01),The expression of TGF-?1 reached maximum at postoperative 3d(P<0.01)that group B is higher than group C,which shows that HRS can make theexpression of liver regeneration advanced genes(HGF,Cyclin-D1)increase,and inhibit theexpression of TGF-?1.Results showed:(1)HRS can effectively promote remnant liver function recovery,promote the recovery of liver tissue,lighten Inflammatory cell infiltration,alleviate the degree of hepatic sinusoidal endothelial cell injury and improve the postoperative liver environment.(2)HRS can accelerate the increase of the concentration of VEGF,Ang-1 and Ang-2.That showed that HRS plays an important role which is regulatory in the secretion of VEGF,Ang-1 and Ang-2 regulation.(3)HRS promotes the expression level of direct mitogen HGF,Cyclin-D1,PCNA increased and inhibits the expression of TGF-?1,which show that HRS promote regulatory role in liver regeneration. |
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