| Objective:Through observing the effects of Kangxian Ruangan granule on nuclear factor-κB(NF-κB) p65mRNA positive rate and patho-logical changes to the ethanol plus CCL4-induced hepatic fibrosis in rat liver tissue,to study therapeutic effect of Kangxian Ruangan granule on hepatic fibrosis,so as to investigate the molecular biological mechanisms of Kangxian Ruangan granule in hepatic fibrosis prevention and treatment,and to provide the theotetical basis for its clinical applications.Methods:40 male SD rats were randomly divided into the following 3 groups:Model control group(M,n=15),Kangxian Ruangan granule group(K,n=15) and normal control group(N,n=10).For M and K group,the rat hepatic fibrosis-model was induced by intraperitoneal injection of 40%CCL4 in olive oil twice a week and 5N ethanol as the only drinking water;the N group was injected the same dose olive oil in the same period.The K Group,in addition to CCL4 and ethanol,rats were given 2g/100g body weight(medium dose) of Kangxian Ruangan granule through stomach tubes every day; M,N group were given physiological saline as comparison over the same period,a total of seven weeks,At the end of the seven weeks,blood and liver specimens were obtained.Right lobes of liver tissues of each rat were taken to 10%formaldehyde and 4% paraformaldehyde fixed,paraffin sections are made.The structural changes of liver tissues was observed by liver tissue biopsy;expression of NF-κB p65mRNA were detected by in situ hybridization;the serum obtained was analysed for liver function tested,including total protein(TP),albumin(ALB),globulin (GLO),ALB / GLO ratio,ALT,AST changes.Results:1.General performance:Rats of model control group were spiritless,lethargy,poor feeding,weight loss and their furs were haggard,lost theirs luster.Rats of normal control group were active,eating well,increasing body weight,their furs were smooth and bright.The state of rats of Kangxian Ruangan granule group were between the two groups.During the experiment,6 rats of model control group were dead,4.rats of Kangxian Ruangan granule group were dead,rats of normal control group all survived.2.Roughly observation:Livers of rats of normal control group appeared dark red,textures were soft,edges were sharp and none abnormal changes,while livers of rats of model control group reduced the size,textures were hard,and some emerged granular nodular surface,edges were blunt;a small number of liver appeared gray,significantly reduced size,with a small amount of ascites,livers of rats of Kangxian Ruangan granule group reduced liver volume,textures were hard,the changes between the normalandthechanges,none obvious nodular surface,the changes were better than the model control group.3.Under the light microscope,HE staining shows that the structure of hepatic lobules of normal control group was obviously complete,the liver cells arranged rule,assumed the radiated distribution around the central vein,in the hepatic lobules there were not cell denaturation,not inflammatory cell infiltration.The hepatic lobules of the model control group were disarrangement of the structures,the arrangement of liver ropes were disorder,the textile fiber knot contracts the organization proliferation,formed the textile fiber gaps,and the gaps encircled and separated normal liver organization obvious fake iobules were formed.But compared with the model control group the pathological changes of Kangxian Ruangan granule group were significantly reduced.The connective tissues were decreased obviously,the textile fiber gaps were obviously thinner,and the obvious fake lobule cound not be seen.4.the liver function examination:In model control group, serum albumin(ALB) obviously reduced,while ALT,AST all went up distinctly(P<0.05);In Kangxian Ruangan granule group,ALB rose, ALT and AST declined obviously,compared with model control group,their differences were remarkable(P<0.05).5.Detection of NF-κBp65mRNA:The expression of model group was significantly enhanced than normal group(P<0.05),The expression of Kangxian Ruangan granule group was significantly reduced than the model group(P <0.05).Conclusion: The experimental results show that the CCL4 plus ethanol can induce the increase of expression of NF-κBp65mRNA in rat liver tissues,promoting the formation of liver fibrosis.Kangxian Ruangan granule can downregulate the expression of NF-κBp65mRNA in rat liver tissues,inhibit the formation and development of liver fibrosis.The results suggest that mechanism of anti-liver fibrosis of Kangxian Ruangan granule may be achieved through regulating the transcription process of NF-κBp65mRNA.
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