Effect Of Radix Isatidis Polysaccharides On Nitric Oxide And Endothelin-1 In Orthopotic Liver Autotransplantation Of Rats

OBJECTIVE:Hepatic ischemia-reperfusion injury (hepatic ischemia reperfusion injury, HIRI) has existed in any surgical procedure of liver transplantation, which is the one of the important reason of primary nonfunction. There are many facts that leads to HIRI, it is generally accepted that the mechanism is related to microcirculatory disturbances, oxygen free radical generation, depletion of energy, calcium overload, dysfunction of mitochondria. In recent years, studies suggest that microcirculatory disturbances play an important role which can determine the final liver tissue injury. NO and ET-1 are two opposite utility of vasoactive substances, the concentration disequilibrium of which is the significance reason for microcirculation disturbances. Therefore, this experiment is based on the model of orthopotic liver autotransplantation of rats. To investigate the effect on the transplanted liver mediated by indigowoad root polysaccharide (IRPS) after the operation and the change of NO, ET-1 content, which may offer the information for clinical practice, lessen postoperative complications, and elevate the postoperative quality of life and the survival rate.METHODS:SD rats were assigned into three groups randomly, including NC group: normal control group (n=32); AT Group: autotransplantation group (n = 32), before the operation the rats were administer distilled water (30mg/kg) through intragastric administration once per day and continued 14 days; IRPS Group: indigowoad root polysaccharide group (n = 32), the rats in this group were received indigowoad root polysaccharide (30mg/kg) at the same method. Blood samples were collected at 1, 6, 12, 24 hours after operation of three groups for examination of the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), and endothelin -1 (ET-1); the micro-structure of liver cell under light microscope and the ultrastructural changes of liver cell mitochondria were determined in three groups.RESULTS:(1) Rats surgical success criteria: Rats instantly awake, stand up, and can maintain an effective breathing, heartbeat. anhepatic phase was controlled in 20±2.2 minutes.(2) The status of the serum ALT and AST: As compared with NC group, the ALT and AST was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05).(3) The status of the serum NO and ET-1: As compared with NC group, the ET-1 was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05). the serum content of NO was decreased in AT and IRPS group, compared with NC group, but the content of NO was more decreased in AT group than that in IRPS group at whole process.(4) The micro-structure of hepatocytes: In postoperative 1h, 6h and 12h the hepatocytes of AT group was severe edema, sinus hepaticus congestion and narrow significantly, endochylema raritas, and mild increase in inflammatory cells. Compared with the AT group, IRPS group of liver cells showed only mild edema. In postoperative 24h, the hepatocytes of AT group was focal necrosis, and periportal lymphocyte infiltration and necrosis; compared with AT group, the IRPS group only show moderate diffuse edema of liver cells, no significant necrosis, and hepatic lobule structural integrity.(5) The ultrastructural changes of hepatocytes and mitochondria: the hepatocytes in the AT group had concentrated; increased electron density as the cells began to condense, condensed or split nuclei and nucleoli, swollen mitochondria, unclear mitochondrial membranes, some disrupted membranes and mitochondrial cristae which became solubilized. Nevertheless, the appearance of hepatocytes in the IRPS group was much better than that in the AT group. The structure of the mitochondria in the IRPS group was normal and was much better than that in the AT group.CONCLUTION:Indigowoad root polysaccharide on hepatic ischemia reperfusion injury has a protective effect after orthotopic liver autotransplantation, which may be related to increase NO levels and lower levels of ET-1 to reduce the microcirculation disturbance.