| 1． ObjectiveHepatic injury was a kind of common pathologic change of more1iverdiseases．Its long term existence was the importantly initiating agent of hepaticfibrosis，cirrhosis and he pa to car c i no genesis．So，treating hepatic injury was themail point of curing clinical liver ailment．Although mechanism of Hepatic injurywas complex，Numerous studies show that medicine can achieve the protectiveeffect of liver damage in a variety of ways, with the majority of its mechanism ofantagonism toxins, improve hepatic microcirculation, inhibition of hepatocyteapoptosis, reducing inflammatory cytokines and free radical scavenging and otherrelevant. Therefore the use of integrated traditional Chinese medicine treatment, toreduce or control the occurrence of liver injury, should be an effective and feasibletreatment of both ideas.SND (si n i decoction, SND) from "Treatise on", the original party has the effectof Yang deficiency, mainly Sh eng little chill in the treatment of certificates yangchill. Modern pharmacological studies have shown that: SND has strong free radicalscavenging, antioxidant, improve microcirculation, immune regulation, anti-tumorand other pharmacological effects. Our group mainly use their immunomodulatoryeffects to observed the effect of Modified Si n i Decoction in the treatment of acuteinflammation.Therefore, based on clinical experience of my ad visor, the subject on the basis ofthe original parties to join dogwood. have antioxidant, immunomodulatory, anti-inflammatory medicine constituted to regulate immune function-based flavoredSND. In recent years, mice with acute liver injury model used concanavalin A (ConA) replication by T lymphocyte-mediated screening for experimental drug studies, aswell as the mechanism of cell biology and molecular biology of liver cell injury idealanimal model. the experiment by concanavalin A copy of acute liver injury model,using flow cytometry, immunohistochemistry, ELISA and other methods to observethe protective effect of Jia wei Si n i decoction on immunological liver injury, and toexplore its protection mechanisms to provide a theoretical basis for clinical use.2.MethodThe60healthy female NIH mice were randomly divided into six groups: normalsaline (NS) in the control group, Con A model group, high dose group SND flavored,flavored SND dose group, low-dose flavored SND group. Using intravenousinjection of Con A copy of acute liver injury model, all of the mice were fasted for12h after the removal of the eye to take blood and liver of mice in each group todetect liver index, serum was measured by ELISA method aspartateaminotransferase (AST), alanine aminotransferase (ALT) activity and cytokinessuch as tumor necrosis factor in mice (TNF-a) levels and the use of HE staining ofliver histopathology morphology observed in liver tissue Fas antigen expression byimmunohistochemistry. By FACS Cali bur flow cytometry in peripheral blood CD4+, CD8+T cell subsets and activation status. All data x±s table; efficacy of eachtreatment group comparative analysis of variance with repeated measures data;comparative analysis between groups ANOVA; all data processing using SPSS16.0software, P <0.05as significant.3.Results:3.1The mice were observed generallySaline control group of mice easily, good spirits, food, hair normal, waterquantity, urine normal; Con A model animal group showed hair reduction, slow,apathetic, lying curled, fatigue, anorexia, yellow urine, loose stool. The positive druggroup and flavored SND large, medium and small dose group of mice are also partof the above symptoms and signs, but the occurrence of cross-model group delay, and with increasing doses of the drug, the symptoms and signs gradually reduced.3.2Protective effect of Jiawei Si n i decoction on Con A immunological liver injuryin miceafter intravenous injection of Con A modeling, the activity of model group serumtransaminases elevated levels of ALT and AST, visually dissected liver volumeincreases, the film swelling, blunt edges, dark red, visible on the surface punctatehemorrhage and necrosis flaky points, there was a marked congestion of spleenenlargement. Mouse model of liver index was significantly higher than the salinecontrol group, and the difference was significant statistically significant (p <0.05).Microscope lobular architecture model mice damage, blurred, seen most of the livercells appeared punctate, small pieces or diffuse necrosis, necrotic areas within alarge lymphocytes and mononuclear cell infiltration, a majority of the cytoplasmphagocytic cell debris and lipofuscin Kupffer cell hyperplasia;Moreover cytoplasmicswelling most hepatocytes, osteoporosis,and some were ballooning, and seen manyround eosinophilic hepatocyte apoptosis. The treatment group close to the normallobular architecture, hepatocyte necrosis and inflammatory cell infiltrationdecreased. These results show that the mice after injection of Con A, inducedsignificant liver damage. The flavored SND large and small-dose group had toreduce the activity of serum transaminase levels, and can significantly reduce thedamage immunological liver injury in mice lobular structure of the liver tissue, livertissue to reduce inflammatory cell infiltration, reduced liver cell degeneration, theformation of necrosis and apoptosis, the degree of Con A-induced liver injury has atherapeutic effect, and a clear dose-effect relationship. The results of Jia wei Si n idecoction immunological liver injury provides an experimental basis, the next stepwe do further research on its mechanism of liver.3.3Jiawei Si ni decoction on immunological liver injury in mice peripheral bloodCD3+, CD4+, CD8+impactThe results show that the model group mice blood CD+4, CD+8ratio significantlydecreased compared with normal control group, compared with normal group therewas a significant difference (P <0.05); compared with the model group, the treatment group can cause small rat whole blood CD4+、CD8+ratio was significantly higher (P<0.05), described Sini plus dogwood soup can significantly up regulated in liverinjury has been significantly reduced whole blood in mice CD4+、CD8+ratio,improving peripheral blood T lymphocyte subsets ratio, reducing its infiltration ofthe liver, thereby reducing its cytotoxic effects on liver cell damage.3.4The Effect of Jiawei Si n i decoction on liver injury induced cell apoptosisThe results showed that the levels of serum TNF-a levels were increasedcompared with the control group, the drug group were P <0.05. Model group Fasantigen abundantly expressed in the liver cell membrane, was significantlyespecially in piecemeal necrosis and necrosis of lymph node or bridge small roarinfiltration area around the liver cell membrane. Control group and the drug groupserum TNF-a levels changed little, hepatocyte F as antigen expression is notobvious. According to the description of Jiawei Si ni decoction can interfere with theexpression of Con A mouse liver tissue F as antigen and serum TNF-a levels ofapoptotic cells in acute liver injury have better protection.4.ConclusionJia wei SND can active peripheral CD4+、CD8+T cells and reduce cytotoxic Tlymphocyte infiltration into the liver; suppress the expression of murine serum tumornecrosis factor and hepatocyte Fas antigen, thereby blocking off apoptosis achievethe purpose of anti-immune liver injury. This study provides new ideas for theprevention and treatment of liver injury in medicine in the future, and lay the initialfoundation for further development and research of traditional prescriptions. |
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