| Objective: Hepatic fibrosis(HF) is caused by a variety of pathogenicfactors (alcohol, viruses, drugs, and autoimmune, etc.) under the action ofchronic liver injury and liver repair and healing, to the development ofcirrhosis a dynamic process and common pathological pathway, which ischaracterized by the abnormal proliferation of fibrous connective tissue in theliver, liver structural damage within fibrosis of sinusoidal capillarization withthe hepatic lobule, at the same time can cause liver dysfunction and portalhypertension; nature of the cells The extracellular matrix (Extra-cellularmatrix ECM) synthesis and decomposition imbalance, ECM excessivedeposition. Numerous studies have demonstrated reversible liver fibrosis stage,to arrest and reverse liver fibrosis and can greatly reduce the incidence ofcirrhosis and liver cancer. The activation of hepatic stellate cells (HSC) is thecentral link of the occurrence and development of liver fibrosis, and manycytokines involved in the activation of hepatic stellate cells, such as: theconnective tissue growth factor (CTGF), platelet-derived growth factor(Platelet derivative growth factor, PDGF), transforming growth factor-β(TGF-β), the HSC is activated and transformed into myofibroblasts (MFB) toexpress a variety of cytokine receptors, further proliferation, synthesis andsecretion of a large number of ECM, promote collagen production, deposition,leading to the occurrence of liver fibrosis. Scholars on the pathogenesis ofliver fibrosis large number of studies, but the treatment of liver fibrosis still noeffects of drugs, with the state in recent years the importance of thedevelopment of Chinese medicine, Chinese medicine against liver fibrosistreatment shows larger potential.Biejiajian pill from the Eastern Han physician the Zhongjing book "the Golden Chamber malarial abnormal pulse and governance",with Endo-metriosis, circulation meridians effect. This experiment by the the classiccarbon tetrachloride (CCL4) modeling method to establish the model ofhepatic fibrosis, drug intervention model change observed in experimentalanimals generally detect serum laminin protein (LN), type IV collagen (Col-Ⅳ), serum alanine aminotransferase (ALT), aspartate aminotransferase (AST),serum albumin (ALB) content, observed liver pathology morphologicalchanges, to detect liver tissue CTGF level, P38, TIMP-1, MMP-9levelsdetected by these indicators, research on the treatment of liver fibrosis and thepossible mechanism of action for clinical treatment to provide experimentalevidence.Methods:56healthy purebred clean SD male rats, weighing360-410g,provided by the Experimental Animal Center of Hebei Medical University,adaptability fed for one week, were randomly divided into6groups, includingnormal group (A), model group (B), the western medicine group (C), Chinesemedicine low dose group (D), the middle dose group (E), the high-dose group(F). In addition to the normal group, the rats in each group were injectedsubcutaneously in experiment40%CCl4olive oil mixed solution5ml/kg of3ml/kg, normal group, subcutaneous injection of the same amount of oliveoil injection every3days thereafter, at the same time, traditional Chinesemedicine and western medicine daily were given traditional Chinese medicineis low in the high dose group rats Biejiajian pill0.55g/(kg.d),1.1g/(kg.d),2.2g/(kg.d), colchicine0.1mg/kg orally. Model group were given distilledwater. Experimental fourth weekend the week randomly sacrificed model rats,light microscopy of liver pathological changes in the first six weeks of theexperiment successful modeling. After the modeling, given to the normalgroup, model group, rats in each group and the western medicine group therats continue gavage five weeks, once a day, the dose based on body weight.11weeks of the experiment, rats were fasted for12hours, and heart bloodchloral hydrate anesthesia after centrifugation,determination of serum ALT,AST, ALB level; radioimmunoassay of serum LN, Col-Ⅳ level;-80℃until from rat liver tissue, respectively, for HE and Masson staining to observepathomorphological changes of the liver of rats in each group; selected ratliver tissue of the same part of the quick frozen in liquid nitrogen and placedreverse transcription polymerase chain reaction (reverse transcriptionpoly-merase chain reaction RT-PCR) assay in liver tissue levels of CTGF andTIMP-1MMP-9; immunohistochemical methods to detect the expression ofP38protein in the liver tissue.Results:1General condition of experimental ratsBody weight of rats during the experiment, the normal group showedsustained growth, usual diet, the eyes of God, shiny fur, reacted swiftly. Modelgroup in western medicine rats in each group general condition is poor,unresponsive, fur bleak, irritability, emaciation. In addition to the normalgroup, the rest of the group were death: model group, two died, Chinese, eachone died in the high-dose group, western medicine group.2The pathology changes of hepatic tissue in each groupLight microscope normal rats liver mainly to liver cells, liver cells werecord-like arrangement, arranged radially around the central vein, hepaticlobule structural integrity of the liver cells arranged in neat rows. Rathepatocyte swelling degeneration model group, was part of the ballooningdegeneration of cells ill-cytoplasmic uneven or intracytoplasmic vacuoles,interlobular thick fibrous septa, lobular architecture destruction, around thecentral vein behold few spotty necrotic cells, accompanied by neutrophils,monocytes and other inflammatory cell infiltration; hepatocyte nucleardeformation or slightly pyknotic marked hyperplasia of fibrous connectivetissue of the portal area, formation pseudolobules. Chinese medicine treatmentgroup, although liver cell damage, structural damage, but significantly reducedcompared with the model group, the formation of false lobules less structure isclearer, the liver sinusoidal mild expansion moderate hyperplasia of portalarea and fibrous connective tissue. Colchicine treatment group, the degree ofliver damage changes between model group and Chinese medicine group between the treatment groups. Description Biejiajian pill to relieve the effectof liver fibrosis and significantly better than western medicine group.3The serum markers of different groupsThe results showed that: compared with the normal group, the modelgroup, colchicine group, Chinese medicine low, the middle dose serum ALT,AST content compared with the normal group was significantly higher (P<0.05), traditional Chinese medicine high dose group and normal group, ALT,AST content no difference (P>0.05); each dose group of traditional Chinesemedicine and western medicine group compared with the model group, serumALT, AST content decreased significantly (P <0.05); compared, traditionalChinese medicine and western medicine group dose serum ALT and AST weresignificantly reduced (P <0.05), traditional Chinese medicine in the dosegroup with which it has no differences (P>0.05); between traditional Chinesemedicine in the treatment groups compared to serum ALT, AST levels fromlow to high order: high-dose group, middle dose group and low dose group,the statistics showed that Chinese medicine among the treatment groups therewere significant differences (P <0.05).The serum ALB content of results: Compared with the normal group,Chinese medicine treatment of high-dose group with which it has nosignificant difference (P>0.05), model group, western medicine group,Chinese middle dose group and high dose group significantly decreased serumALB content, statistically significant compared with the model group (P<0.05); Chinese medicine high dose group, middle dose group, serum ALBcontent of the western medicine group was significantly higher than the modelgroup was statistically significant (P<0.05), traditional Chinese medicinetreatment of low-dose group and the model group, the content of ALB was nodifference (P>0.05); compared with the western medicine group, Chinesehigh-dose group, serum ALB were significantly higher (P<0.05), traditionalChinese medicine in the dose group compared serum ALB content differencenot statistically serum ALB content of significance, the low-dose groupreduced (P<0.05); pairwise comparison between Chinese medicine each dose group, serum ALB level from low to high order of: low-dose group, middledose group and high dose group, and each The difference between the twogroups was statistically significant (P<0.05).4The levels of Col-IV and LN in the serum of different groupsCompared with the normal group, the differences in Chinese medicine highdose group and the normal group was not statistically significant, theremaining serum LN, Col-IV were significantly increased (P<0.05); comparedwith the model group, each dose of the traditional Chinese medicine group,western medicine serum LN, Col-IV content was significantly lower than themodel group (P<0.05); compared, traditional Chinese medicine and westernmedicine group reduced serum levels of high-dose group was statisticallysignificant (P<0.05), traditional Chinese medicine in the dose group serum LN,Col-IV content with which it has no significant (P>0.05); Chinese medicinebetween each dose group compared to serum LN, Col-Ⅳ levels ranging fromlow to high order: Herbal high dose group, middle dose, the low-dose group,and the difference was statistically significant (P<0.05) between each dosegroup.5The expressions of CTGFmRNA, TIMP-1mRNA, MMP-9mRNA in thehepatic tissue of different groups5.1The expression of the CTGFmRNA, TIMP-1mRNAThe results showed that: compared with the normal group, the modelgroup, western medicine group, Chinese medicine content of each treatmentgroup were significantly increased (P<0.05); compared with the model group,western medicine group, we reduce the expression of traditional Chinesemedicine in the treatment groups (P<0.05reduced); compared, traditionalChinese medicine and western medicine group, high dose group expressionincreased expression of traditional Chinese medicine low dose group werestatistically significant (P<0.05), compared to the traditional Chinese medicinein the dose group and western group was not statistically significant (P>0.05);pairwise comparisons between traditional Chinese medicine each dose groupwas statistically significant (P<0.05) 5.2The expression of MMP-9mRNAThe results showed that:: Compared with the normal group, model group,western medicine group, the expression of traditional Chinese medicine in thetreatment groups were lower (P<0.05); compared with the model group,western medicine group, the content of the expression of traditional Chinesemedicine treatment group were lower (P<0.05); compared with the westernmedicine group, high dose group content in traditional Chinese medicineelevated expression was statistically significant (P<0.05), the low-dose groupcontent in traditional Chinese medicine express higher statistically significant(P<0.05), traditional Chinese medicine in the dose group compared to thewestern medicine group difference was not statistically significant (P>0.05);Chinese medicine between each dose group pairwise comparisons werestatistically significant (P<0.05).6Immunohistochemistry assay in P38protein expressionP38protein expression was brown, weakly positive expression in normalrat liver tissues; compared with the normal group, the protein expression wassignificantly increased (P<0.05); compared with the model group, westernmedicine group, Chinese medicine treatment of histone lower expressionlevels (P<0.05); compared with the western medicine group, Chinesemedicine high dose group protein expression was lower (P<0.05) and proteinexpression of traditional Chinese medicine in the dose group and westerngroup difference was not statistically significant (P>0.05) TCM low-dosegroup protein expression is higher than the western medicine group, thedifference was statistically significant (P<0.05); pairwise comparison betweenChinese medicine each dose group, the amount of protein expressiondifference was statistically significant (P<0.05), high-dose histone expressionlevels close to the normal group.Conclusions:1Rats with liver fibrosis in rats presence of liver dysfunction andexcessive deposition of extracellular matrix. Biejiajian pill can significantlyreduce liver damage and hepatic fibrosis degree, has the effect of improving liver function2Biejiajian pill can significantly reduced the expressions of CTGF,TIMP-1, reduce the effect on inhibition of MMP-9TIMP-1, thus contributingto the degradation of the extracellular matrix, and to reduce the synthesis ofextracellular matrix improve liver of their one of the mechanisms of fibrosis.3Biejiajian pill of Chinese medicine reduce the expression of the P38protein, inhibit the cell proliferation and differentiation, so that blocking theprocess of liver fibrosis, this may be one of the mechanisms of anti-fibrosis. |
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