To Investigate The Effect Of Pricking Spilled Blood Therapy On The Apoptosis Of Hepatic Stellate Cells And The Expression Of Bcl-2 And Bax In Hepatic Fibrosis Rats

Bloodletting therapy is an important part of Traditional Chinese Medicine, belonging to acupuncture therapy, which is called bloodletting puncture or qimai in ancient and pricking blood therapy now. The graduate advisor of author found that bloodletting therapy had obvious efficacy on Hepatic Fibrosis in clinical practice.The related literature revealed that acupuncture have achieved satisfactory effects in the prevention and treatment of hepatic fibrosis. Based on the clinical experience of tutor and exiting research, the studying team designed a experiment to explore the effects of bloodletting on liver fibrosis of rats model induced by CCL4. In this experiment, the collaterals on or near LV3-Taichong, ST36-Zusanl and SP9-Yanglingquan of the rats of HF were pricked with a three-edged needle.We emphatically observed the pathological change of the liver,HSC apoptosis in liver and expression of apoptosis-regulating genes(Bax/Bcl-2). At last, the mechanism of bloodletting in prevention and treatment of HF rats will be discussed. We hope to open a new way to teat HF.This paper mainly includes two parts of the literature and experimental research.Firstly, the literature researchThe literature research include three reviews. The first literature review provides a summary of bloodletting development. We will analysis the characters and innovation of bloodletting therapy in different period. In the second literature review,we will discuss the understanding of TCM on HF and summarize the advantages and disadvantages of TCM on the prevention and treatment of HF. At last,we will provide a prospect of TCM on treatment of HF.The last literature review will summarize the progress of HSC apoptosis.We mainly investigate the genes, proteins, cytokines and TCM factors, which control HSC apoptosis, and find the strengths and potential of TCM on controlling HSC apoptosis.Secondly, the experimental researchObjectiveIn this experiment,we induced HF of SD rats model by CCL4. The collaterals on or near Taichong, Zusanl and Yanglingquan of the rats of HF were pricked with a three-edged needle to treat the rats. We observed the pathological change of the liver,HSC apoptosis in liver and expression of apoptosis-regulating genes (Bax, Bcl-2). So, we can explore the mechanism of bloodletting in prevention and treatment of HF rats.MethodsForty-eight male SD rats were randomly divided into normal control group、 model group、colchicines-controlled group and bloodletting group with 12 cases in each group. Except the normal control group, the other three groups were induced into liver f ibrosis model with 40% CCL4 diluted by peanut oil. After 6 weeks, the colchicines-controlled group was given colchicine 200μg/kg/d with 5 times a week;the blood-letting group had 0.15-0.2ml hemorrhage on or near Taichong, Zusanl and Yanglingquan with 2 times a week. After 12 weeks, we observed the general conditions of rats; Small piece of liver tissue selected from left lobe of liver was embedded in paraffin, sliced and stained with Masson and observed its Pathological changes; the apoptosis of HSC was observed by TUNEL and the expression of Bcl-2 and Bax wan tested by PCR.ResultsThe general conditions:Rats in normal group were generally in good condition, but the rats in model group had retardation,poor diet,weight loss, yellow and dry fur and poor mental status. The condition of rats in colchicines-controlled group and bloodletting group were better than that in model group.The morphological change of liver tissue:Masson staining showed structural integrity of hepatic lobule. There was only a little collagen fibers in portal area.Compared with normal group,Masson staining result indicated significant liver fibrillation. The fibrillation of colchicines-controlled group and bloodletting group wan slighter than that in model group.It indicated the treatment was effective.The HSC apoptosis:Compared with normal group, the other groups’number of HSC apoptosis increased; The number of HSC apoptosis in both the colchicines-controlled group and blood-letting group were more than the model group(P<0.05, P<0.01); Compared with colchicines-controlled group, the HSC apoptosis of blood-letting group increased(P<0.05).The expression of Bcl-2 and Bax:Compared with model group,the expression of Bcl-2 decreased in both the colchicines-controlled group and blood-letting group and the difference was significant(P<0.05). On the other hand, the expression of Bax increased slightly.The results indicated that bloodletting therapy could significantly decrease the expression of Bcl-2,but increased slightly increase the expression of Bax.ConclusionFirstly, Bloodletting therapy has great treatment effects on HF rats and can reduce the fibrillation, which may be connect with the increase of HSC apoptosis.Secondly, Bloodletting therapy can significantly decrease the expression of Bcl-2,but increase slightly increased the expression of Bax.Thirdly. Bloodletting therapy can treat HF of rats effectively. The mechanism may be connected with the increase of HSC apoptosis and Bcl-2 expression and decrease of Bax expression.