| Background:Hepatic diseases play a significant role in the health and living quality of human beings. Fibrosist is a positive wound-healing response that organism reacts to chronic liver injury. It is also common pathological basis of quantity of liver diseases. It is conformed that fibrosist can be reversed, However, once developed into liver cirrhosis, It is rearlly hard to revers. Thus,interception and reversion of liver fibrosis are regarded as key points of prevention and therapy for chronic liver disease and cirrhosis. Based on developmental biology of the fascia like connective tissue, combined with traditional chinese medicine of research meridians,modern acupuncture basic theory and the virtual human research results, some scholars proposed functional systems of human existence that is the tenth self-supervision and surveillance system. The theory is that fascia system may be the anatomical basis of meridian, where "point" is the fascia bracket can accept and generate strong biological information to stimulate the nerve receptors and activity of cell aggregation Department,"meridian" to "point" is fascia structure connected between the anatomical structures or afferent nerve close to.To investigate the acupuncture treatment of liver fibrosis in the cellular compartment mechanisms, we choose human embryonic stem cells (hESCs) as a tracer cell. so take the advantage of multiplex differentiation of hESCs,using hESCs as a tracer cells to transport back to a healthy SD rats implanted,Liver fibrosis model by copying, testing conditions in the acupuncture intervention fascia cell differentiation within the tracer migration path in the idea of an experiment study.Objective:Adopt electric acupuncture on acupuncture point and fascia,Then Observe the distributing and differentiating Migration traces of marked hESCs which were transplanted into the body of Liver fibrosis model rats. Discuss the possible cytology mechanism of electric acupuncture on acupuncture point and fascia.Methods:1.96male SD rats,weighting160-180g, were randomly divided into normal control group, normal control with hESCs transfusions group,hepatic fibrosis model group, hepatic fibrosis model with hESCs transfusions group, QI MEN points acupuncture with hESCs transfusions group,YANG LING QUAN points acupuncture with hESCs transfusions group, QI MEN connective tissue acupuncture with hESCs transfusions group and YANG LING QUAN connective tissue acupuncture with hESCs transfusions group. Every group has twelve rats. Except for normal control group,others were fed white wine、lard oil and pyrazole mixture to established animalmodels of alcohol liver fibrosis, the normal control group were fed saline daily.2. marked hESCs with Ad5-EGFP, then were observed under fluorescence microscope.3. hESCs marked by Ad-5EGFP were transplanted into the body of alcoholic liver fibrosis model by caudal vein.4. Acupancture the blood serum of Qi Men、Yanglingquan and relative connective tissue intensive area for20minites every two days, course of the treatment is21days.5. After established model and acupuncture,we use20%urethane to anesthesia the rats,take out the liver tissue and arterial blood.liver was immediately make tobe frozen sections to observe the transmission rate of labeled cells under fluorescence microscope and observe the distribution of marked stem cells by general microscope.Results:The liver fibrosis rats model we established was proved tobe successful by blood serum markers of liver fibrosis、HE staining and Immunohistochemistry(IHC). After labeled by Ad5-EGFP, hESCs sent a green light excited by blue fluorescence after labeled48hours.the rate of labeled cells is up to90%.Shape of the cell is clearly visible. Attached cell is with a spindle shape or triangle,there are many fluorescent particles in hESCs. every treatment group rats was proved that the degree of liver fibrosis all improved on different scale by blood serum markers of liver fibrosis、HE staining and IHC.By comparison QI MEN points acupuncture with hESCs transfusions group,YANG LING QUAN points acupuncture with hESCs transfusions group, QI MEN connective tissue acupuncture with hESCs transfusions group and YANG LING QUAN connective tissue acupuncture with hESCs transfusions group, there is no statistical differences between acupuncture point and fascia.Conclusion:hESCs were successfully cultured and the rats model of alcoholic liver fibrosis is established in this experiment. electric acupuncture with the transplantion of stem cells are effective treatments to prevent and cure liver fibrosis. Ad5-EGFP recombinant adenovirus is a fairly good vectors to infect hESCs,is simple and efficient.the cytology mechanism of the hESCs transplantion May have relations with micro environment hESCs in.which was Induced or improved by electric acupuncture. the detailed cytology mechanism Waits For Further Studying. |
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