The Protective Effect Of Xuebijing To Renal Injury Of Portal Hypertension Model Rats After Anhepatic Phase

Objective: Nearly half a century, relying on modern surgical techniques, organ preservation and Peri-operative management of the rapid development of technology, to the advances in transplantation immunology and molecular biology as a backup, liver transplant for end-stage liver disease had become the only effective means. Most receptors in China suffered from posthepatitic cirrhosis with portal hypertension and a lot of enlarged portosystemic collaterals. Based on this characteristic, the orthotopic liver transplantation(OLT) without venous bypass, which was simpler and cheaper relatively, had a great prospect in China . However, with the absence of venovenous bypass during one hour of anhepatic phase, there will be an ischemia/reperfusion renal injury. Acute renal dysfuncion after liver transplantation was a risk factor."Xuebijing"was famous for Emergency Medicine by Professor Wang today over 30 years of scientific research into clinical practice in the National Priority new drugs. This study was designed to investigate Xuebijing injection in the treatment of portal hypertension rats after anhepatic phase protective effect on the kidneys, for clinical acute renal dysfuncion after liver transplantation on finding a new drug. Methods: 110 healthy male Wistar rats, weighted 250-300g, 15 were randomly selected, 10 as a sham operation group (SOG), 5 were used to justify portal hypertension models. The recoverable prehepatic portal hypertensive models were generated on the other rats through the first operation which partly narrow portal vein and let the mark ring in, three weeks after the establishment of portal hypertension model (PHM), from only five were used to justify portal hypertension model. Collecting five normal rats and rats with portal hypertension measured free portal pressure and the area of esophageal submucosal veins. Portovenography were performed in normal rats and rats with portal hypertension. The remaining 90 rats in a second-stage operation was performed to identify and remove mark ring after the first ring so that portal reexpansion. Later the same time blocking the first after the inferior vena cava and hepatic portal, 60 minutes after opening, According to reperfusion after collecting specimens were divided into six hours of ischemia and reperfusion (I/R6 h), 12 hours (I/R12 h), 24 hours (I/R24h), Xuebijing six hours (XBJ6h), 12 hours (XBJ12h) ,24 hours (XBJ24h), ulinastatin six hours (UTI 6h), 12 hours (UTI 12h), 24 hours (UTI 24h), a total of nine. Made from the inferior vena cava blood serum ALT, AST, BUN, Cr measured evaluation of liver and kidney function; Morphological changes of liver and kidney were observed under optical microscopy,the ultrastructure of renal tissue were examined by transmission electron microscopy also;the changes of SOD and MDA concentrations of renal tissue were studied. All data of test were analyzed by statistic software.Results: Three weeks later after the first operation, all the portal hypertensive model rats were generated successfully. Ascites and varices of viscera could be seen in the PHM rats, but the color and texture of liver were normal. The free portal pressure of PHM rats was 15.6±3.1mmHg,was obviously higher than that of normal rats(7.7±1.7mmHg), and the difference was significant(P <0.05).When the mark ring was removed, the portal pressure was 13.4±2.3mmHg. The portovenography of PHM rats showed that development of liver was slow, and a stenosis could be been at the portal vein trunk proximal to the bifurcation, the vessels of portal system twisted and dilated, the enlarged veins such as left adrenal vein and azygos vein could be seen, the superior and inferior vena cava through the portosystemic shunt developed, the stenosis of portal vein disappeared after removing the mark ring. There were no shunts between portal and benacaval system in normal rats. The area of esophageal submucosal veins of normal rats and PHM rats were 5.620±1.301μm 2 and 33.582±6.425μm 2 respectively, the difference was significant(P <0.05). The level of ALT and AST of I/R group in serum significantly increased at 6 hours, reaching their peak at 12 hours. I/R group compared with SO group, ALT and AST were significantly increased (P <0.01);XBJ group compared with I/R group, ALT and AST were significantly lower (P <0.01); XBJ group compared with UTI group, ALT, AST was no significant difference (P> 0.05); The level of BUN and Cr of I/R group in serum significantly increased at 6 hours, reaching their peak at 24 hours. I/R group compared with SO group, BUN and Cr were significantly increased (P <0.01);XBJ group compared with I/R group, BUN and Cr were significantly lower (P <0.01); XBJ6h group compared with I/R6h group, BUN and Cr were lower (P <0.05); XBJ12h,XBJ24h group compared with I/R12h ,I/R24h group, BUN and Cr were significantly lower (P <0.01); XBJ group compared with UTI group, BUN, Cr was no significant difference (P> 0.05). MDA in the kidney tissues significantly increased at 6 hours, reaching their peak at 24 hours. I/R group compared with SO group, MDA was significantly increased (P <0.01); XBJ group compared with I/R group, MDA was significantly lower (P <0.01); XBJ group compared with UTI group, MDA was no significant difference (P> 0.05); SOD with MDA contrary. Optical microscope renal proximal tubular epithelial cells of R/I group was swelling. Karyopyknosis, nuclear fragmentation and endochylema concentration could be seen cell. Cell debris and renal tubular epithelial cell necrosis could be seen in lumens intracavitary. Cast could be seen collecting duct. Renal tubule lumens expand and brush border became shorter.XBJ group renal injury was reduced significantly. Renal tubular epithelial cell necrosis was ligher ,some defluvium into renal tubule intracavitary. Optical microscope liver cells edema was Severity in R/I group, some degeneration and necrosis; XBJ group edema, necrosis of the liver cells reduced. TEM observation of the ultrastructural changed in proximal tubules, R/I group cytoplasm was mild edema, and cellular ridge ranked slightly disordered. Chondrosome a few or more cristae and emma coalesced or disappeared. Rough endoplasmic reticulum had serious degranulation, and significantly reduced the number of microvilli.XBJ group proximal convoluted tubule injury reduced. Cytoplasm also could be seen edema. Chondrosome cristae and emma coalesced or disappeared. Rough endoplasmic reticulum had slight degranulation,and the number of microvilli reduced slightly.Conclusion: The methods of partial portal vein ligation to induce the model of prehepatic portal hypertension in the rat was simple, duplicated, reliable and its hypertension was recoverable,it could be applied for the studies on OLT. Renal injury of portal hypertension model rats after anhepatic phase was most serious at 12-24 hours after reperfusion. Xuebijing injection could effectively protect renal injury of portal hypertension model rats after anhepatic phase . At present, we still could not prove that Xuebijing injection protect renal injury of portal hypertension model rats after anhepatic better than ulinastatin.