The Research Of Pharmacodynamics For The Treatment Of Hepatic Fibrosis In Rats,with The Prescriptions For Tonifying Qi,Dissipating Stasis And Removing Toxin

? ObjectiveThis experiment mainly study the pharmacological effect with prescriptions for tonifying qi,dissipating stasis and removing toxin,to explore its action mechanism for the treatment of liver fibrosis?HF?in rats,and provide theoretical basis for Chinese medicine development and clinical application??MethodFirstly,to make a HF model,Wistar rats were injected subcutaneously with mixture of 40%CCL₄ and olive oil?5ml/kg,twice per week?.8 weeks following the first CCL₄ injection,rats were analyzed for Hyaluronic acid?HA?,Laminin?LN?by radioimmunoassay.Six rats were sacrificed and histological assessment for hepatic fibrosis by hematoxylin and eosin?HE?staining and Masson staining.Secondly,to study pharmacodynamics for the treatment of hepatic fibrosis in rats,with the prescriptions for tonifying qi,reducing blood stasis and eliminating toxins?YQZYJD?.Rats which were exposed to CCL₄ for 8 weeks were randomly divided into 5 groups and including YQZYJD high dose group?n=14?,YQZYJD medium dose group?n=14?,YQZYJD low dose group?n=14?,Fuzhenghuayu capsule?FZHY?group?n=14?,Model group?n=14?.YQZYJD high dose group was fed with YQZYJD 22.8g?crude drug?/kg body weight,YQZYJD medium dose group was fed with YQZYJD 11.4g/kg and YQZYJD low dose group was fed with YQZYJD 5.7g/kg.FZHY group,as a positive control group,was fed with FZHY 0.4g/kg.Model group and blank control group were fed with saline daily.After treatment for 8 weeks and fasting 12 hours,rats serum were evaluated for Hyaluronic acid?HA?,Laminin?LN?by radioimmunoassay and some cytokines such as Transforming growth factor-?1?TGF-?1??Connective tissue growth factor?CTGF??Platelet derived growth factor-??PDGF-??were analyzed by enzyme-linked immunosorbent?ELISA?.All rats were sacrificed for histological assessment for hepatic fibrosis by hematoxylin and eosin?HE?staining and Masson staining and Immunohistochemical evaluation fora-Smooth muscleActin?a-SMA?which is a marker of Hepatic stellate cells?HSCs?activation.? ResultsFirstly,In the process of construction of HF model by CCL₄,ordinary circumstances observation demonstrated that rats mental state and their appetite get poor,Urine turn yellow,fur turn get rough and yellow,even local furs have fall off,Individual appear local skin ulcers.The appearance of liver tissue that Color is dark red or gray and the surface is rough,liver appear edema and nodulars,Liver edge become dull.Liver HE staining:Liver cell appear edema and steatosis,Capillary bile duct have cholestasis,Livers Portal area expanded and fibrous tissue hyperplasia,liver tissues most visible collect abbacy fibrosis?PF?,with manifold-send pipeline bridge fibrosis?P-P?and manifold-central bridging fibrosis?P-C?.Compared with blank group,model group have higher content between HA and LN?p<0.05?.Secondly,the spirit,appetite and the superficial knewlege luster of treatment group is improved in the model group,liver tissue have rough uneven texture and apophysis nodules of model group after treatment,liver tissue appear local small nodules,or size of nodules,with no apparent apophysis,treatment group is improved compared with model group.Liver fibrosis grading display class 1 in the high dose of the prescriptions for tonifying qi,reducing blood stasis and eliminating toxins.Hepatic fibrosis,mean score of SSS score is lower than those of other treatment group and model group,trend is The prescriptions for tonifying qi,reducing blood stasis and eliminating toxins can reduce the degree of liver fibrosis.Other indicators?Such as:HA?LN?TGF-?1?CTGF?PDGF-???-SMA?statistical analysis of no significance?p>0.05?,the main problems of the results In this study are as follows:?1?This experiment uses the subcutaneous injection method,absorption process is relatively slow,drug continues to absorb after stop making model,so as to make the degree of liver fibrosis in the further;?2?A handful of serum specimen appeared hemolysis;?3?Due to the lack of time and money,we just use one detection method instead of a variety of methods to refer and analyse,when we detecting TGF-?1?CTGF?PDGF-???-SMA;?4?Some rats died during the process of making model and the treatment,leading to insufficient sample size.? ConclusionsFirstly,Rats model of hepatic fibrosis was constructed successfully by injection subcutaneously with mixture of 40%CCL₄ and olive oil?Secondly,research trend is the prescriptions for tonifying qi,reducing blood stasis and eliminating toxins can alleviate hepatic fibrosis.