The Effect Of Ulinastain On Rat Hepatofibrosis

BackgroundHepatofibrosis is a pilot process between hepatitis and hepatic carcinoma.Hepatic stellate cell activation is the key link with the process of hepatofibrosis.Hepatic stellate cell affected by cytotine, growth factor, chemotactic factor,productions of oxidative stress, which can form a net work amongmedium-target-extracellular matrix.Ulinastain is a trypsin inhibitor isolated fromhuman urine. Ulinastai and its metabolite have several effects. Such as inhibition oflysosomal enzyme, stabilizing lysosome membrane, remove oxygen radical, improvemicrocirculation, inhibit cardiac muscle inhibitor. And there also have fewer reportsabout whether it can reverse hepatofibrosis.ObjectivesEstablish rat hepatofibrosis models. Rat hepatofibrosis models were treated withUlinastain. Observe the reverse effect of hepatofibrosis.Method1、 Establishment of experimental model：72rats were randomly divided into fourgroups,18of each group. CCl4group、TAA group and porcine serum groupwere injected0.5ml/100g CCl4（40%）、200mg/kg TAA（0.03%）�'�0.5ml/rat subcutaneous twice a week. Control group was injected0.1ml/kg NaClsubcutaneous twice a week. Serum was collected to examine the level of ALT、AST、HA、SOD、MDA at3、6、9weeks. The liver fibrosis level and SSS scorewere observed with HE stain.2、18rats with hepatofibrosis were divided into2groups, model+ulinastai (9rats),model+physiological saline (9rats), control (3normal rats). Using ulinastain tointervene hepatofibrosis. Compare AST, ALT, HA, SOD, MDA, LN levelchanges after intervene. Rats liver morphology change was observed using HEstain and type-B ultrasonic. TGF-β1、 caspase-3were detected byimmunohistochemistry.ResultsAnimal models establish1. Weight of rats in three groups have not increased evident.2. CCl4group、TAA group and porcine serum group can observe a cloudy granularshape of hepatocyte and fibrous tissue in the portal tract. ALT level of TAA groupwas higher than control group at three time points(P<0.05). While ALT level ofCCl4group and porcine serum group were not obviously increased(P>0.05).AST level of CCl4group was significantly increased, but decreased at6、9weeks.TAA group and porcine serum group have significantly increased level of ASTand HA. MDA level of three experimental group was increased differently at threetime points.3. Hepatofibrosis grading: At three time points, SSS score of three modeling groupshave great significant difference comparing with control group(P<0.05).At9weeks, SSS score of TAA group is higher than other group（sP<0.05）.SSS score ofCCl4group was2.21±0.24.Effect of Ulinastain on hepatofibrosis1. Serum index: After3weeks medication intervene, ALT, AST, SOD, MDA, HAand LN level of C group have improved. Serum index of C group were closer to normal rat than B group. HA and LN level of C group was209.8±17.63、151.5±23.43. There has significant difference compared with B group(P<0.05).2. Instantaneous elastic ultrasound: After3weeks intervene, elastic number of Fgroup changed great,1.50±0.19kPa. It has great significant difference comparedwith before intervene. The elastic of B group have not change evidence comparingwith before intervene(P>0.05).3. HE stain: HE stain show that rats in C group have fewer inflammatory cellsinfiltration in central vein, ballooning degeneration of liver cells, collagenproliferated, hepatic lobules degradation and pseudolobule arise. However, Bgroup have found lots of inflammatory cells infiltration in central vein andremarkable collagen proliferated than C group.4. Sirius red stain: Type Ⅰand Ⅲ collagen fiber of C group were lower than Bgroup.5. Immunohistochemical stain: Great amount and extensive claybank stainedparticles were found in B group liver immunohistochemical stain. In C group,claybank stained particles were diffused in central vein and portal area, TGF-β1、caspase-3positive cell rate were lower than B group.ConclusionTAA can be successfully induced hepaticfibrosis of rat.Ulinastain can reverse TAAinduced rat hepatic fibrosis and alleviate fibrosis degree and collage fiber deposit,inhibit inflammation. It also can improve serum index, white blood cell count,coagulation index. This also provide a new idea and methods for hepatic fibrosisprevention and cure.