Research Of The Method Of Tonifying Qi And Blood And Promoting Blood Circulation By Regulating The NK Cell Activity For Reversing Rats With Hepatic Fibrosis

Objective:This study mainly discuss the methods of traditional chinese medicine upplementing qi, nourishing blood and promoting blood circulation on through the regulation of NK cells to reverse hepatic fibrosis in rats.Method:Part 1:To make a model of hepatic fibrosis of wistar rats. Except 12 normal male waister rats as blank group,the rest of the rats are made the hapatic fibrosis model with 40% CCL4 and olive oil mixture (5ml/kg, twice per week, for 9weeks).After making HF model for 6 weeks, observate the results of liver tissue of HE staining、Masson staining and average optical density of a-SMA. At the 9th week we make sure the HF model of the waister rats is sucessful by examining six executed rats hepatic fibrosis index and detect HE staining and Masson staining and immunohistochemical method to detect liver tissue of a-SMA.Part 2:To detect NK cells activity of Waister rats. After the HF model of Waister rats is made sucessflully, at the tenth week we stop injecting CCL4 and divide 48 rats into four groups including the Fuzhenghuayu capsule group, the interferona2b group, the model group and the blank control group,meannwhile marking marking the rats and treat rats of every group with Fuzhenghuayu capsule, the interferon alpha 2b, the same volume of 0.9%sodium chloride injection.When treating the rats for eight weeks, we stop drug treatment and stop feeding about 12 hours.HA and LN are tested by femoral artery blood.To extract NK cells from anticoagulant blood is to decte NK cells activity through CD107a/CD56 from ratioflow meter. NK cells extracted from Waister rats are incubated with HSC-T6 bought from the company and we can observe the apoptotic HSCs with a microscope,also we can detect the amount of HSCs apoptosis with a flow cytomatry. From the results we can analysis NK cells destruction to HSC-T6. With immunohistochemical method we detect the a-SMA changes in liver tissue from every group and the number of NK cells in the tissue.Results:Part 1:The rats ordinary circumstances observation demonstrated that rats mental state and their appetite get poor,fur turn get yellow and local furs have fall off. The pathological changes of liver tissue:Liver edge become dull,the surface is smooth, local nodulars appear. Livers tissue immunofluorescence staining prove that a large number of depositing collagen fibers exits at the portal area and also extends portal area, meanwhile thick or thin fiber formation form and split hepatic lobule.Compared with blank group,model group have higher content between HA and LN (P< 0.05).Also average optical density of a-SMA of model group is higher than blank group (P<0.05)Part 2:Fuzhenghuayu capsule group and the interferon alpha 2b may obviously improve the HF rats’state of mind and the superficial knewlege luster.The Liver pathological changes of the rats in Fuzhenghuayu capsule group and the interferon alpha 2b group have no obviouse differences, only the liver edge become dull,local coarse granular, no local nodulars appear. But the whole liver of the rats in the model group is bulky nodularity change, and becomes hard, even the part of some liver becomes brown.Hepatic fibrosis indicators suggest the level of HA and LN of the rats in Fuzhenghuayu capsule group and the interferon alpha 2b group is not obviously different,but liver fibrosis level of the rats in the above two groups indexes lower in the model group (P<0.05).The centralizar blood the Average optical density of a-SMA in the Fuzhenghuayu capsule group and the interferon alpha 2b group is lower than in model group.Immunohistochemical result showed the number of NK cells has no difference between Fuzhenghuayu capsule group and the other groups. Compared with the remaining groups, with flow meter the proportion of CD 107a/CD56 is higher in the fuzhenghuayu capsule group (P<0.01), the proportion of blank group and model group is not significantly different. Compared with other groups, the apoptosis rate of HSCs in the Fuzhenghuayu capsule group is significantly higher (P<0.01).Conclusions:Part 1:HF model of rats is made well by the method that CCL4 which is one of chemical abdominal subcutaneous is injected into abdominal subcutaneous.Part 2:The study proves in the process of treatment of HF rats Fuzhenghuayu capsule which has the upplementing qi, nourishing blood and promoting blood circulation function and NK cells activity have a close relationship.