| Mesenchymal stem cells (MSCs),which differentiate from mesoderm mesenchymal during embryonic development, is pluripotent stem cells.MSCs have the differentiation potential of a variety of tissue cells,which includ bone cells, muscle cells, and many other cells. In recent years, studies have shown that bone marrow mesenchymal stem cells also has a strong role in anti-hepatic fibrosis, wich can differentiated into hepatic oval cells (HOCs) and further into functional liver cells and bile duct cells in injred liver of experimental animals, and such a phenomenon is also present in the body of patients who received a bone marrow transplant or liver transplant.Then a large number of scientific Followed,wich proved that both cytokine-induced in vitro and transplantation in vivo can make bone marrow mesenchymal stem cells differentate into active liver cells to participate in the repair of liver damage and reduce the degree of liver fibrosis, participation in the prevention of liver cirrhosis. This maybe the new methods for the trentment of hepatitis and liver fibrosis.Bone marrow mesenchymal stem cells could secrete Varieties of cytokines. Some of these cytokines could stimulate the proliferation of liver cells, and some could inhibit the activation of hepatic stellate cells. Bone marrow mesenchymal stem cells could also secrete matrix metalloproteinase(MMPs) and urokinase-type plasminogen Plasminogen activator(uPA) wich can accelerate the degradation of extracellular matrix and reduce the deposition of collagen fibers.The implantation rate of Bone marrow mesenchymal stem cells in the body of liver fibrosis animal models is much higher than in the body of other liver injury animal models and normal animal models. But the rate of Bone marrow mesenchymal stem cells that differentiate into functional liver cells is very small. Therefore, the function of bone marrow mesenchymal stem cells in the treatment of liver fibrosis is not only the single mechanism that instead of injurded liver cell.Liver fibrosis animal models have two methods to repair the damaged area of liver, endogenous repair and exogenous repair. There in a kind of hepatic oval cells (HOCs),wich is a kind of liver stem cells that can differentiate into functional liver cells. Hepatic oval cells responsible for the physical update of liver cells,and could participate in the repairment of injured liver in pathological cases. The repair like this is the endogenous repair of the liver damage repairment. Bone marrow mesenchymal stem cells tend to migrate to the liver and differentiate into functional liver cells to take part in the repair of liver tissue after transplantated into the body of hepatic fibrosis animal model. The repair like this is exogenous repair in the repair of liver damage.Bone marrow mesenchymal stem cells in the body of liver fibrosis animal models could not only differentiate into functional liver cells but also differentiate into hepatic oval cells,wich showed that the treatment of bone marrow mesenchymal stem cell transplantation in liver fibrosis play a role by both endogenous and exogenous repairment.In this study refusion bone mesenchymal stem cells into the body of liver fibrosis rat model and acupuncture the rat to discover the potential factors that could affect the migration and differentiation of bone mesenchymal stem cells.Objective:1 Selecting the liver as the target organ, from the start of pathological animal models of alcoholic liver fibrosis, we use the methed of transplantation of mesenchymal stem cells (MSCs)that makered by chloromethyl benzamide (CM-Dil) into the body of animal model of hepatic fibrosis, to track the history of expression,differentiation,and migration locus of mesenchymal stem cells (MSCs) in the area of liver and liver connective tissue, and discussion the cellular mechanism study of acupuncture connective tissue in intervene alcoholic liver fibrosis of rats, and discusse cellular mechanisms of acupuncture connective tissue in treating hepatic fibrosis.2 Mesenchymal stem cells were cultured in vitro and induced to differentiate into hepatocytes.Methods:1 Using the whole bone marrow cells adherent to separation training and purify MSCs. We isolated the purification of MSCs by immunocytochemistry and identify the cultured cells expression to CD44, CD71, CD34.2 The rats were randomly divided into normal control group, hepatic fibrosis model group, points acupuncture group, connective tissue acupuncture group, in which the hepatic fibrosis model group, points acupuncture group, connective tissue acupuncture group were fed white wine, corn oil and pyrazole mixture to established animal models of alcohol liver fibrosis, the normal control group were fed water daily. Points acupuncture group were acupuncte qimen point and Yanglingquan point every two day. Connective tissue acupuncture group were acupuncte the connective tissue intensive area next to the Yanglingquan point. 3 Red fluorescent CM-Dil was labeled to the bone marrow mesenchymal stem cells, and the labeling rate was observed under fluorescence microscope. After Bone marrow mesenchymal stem Cells were successfully labeled, we reinfusion it through the tail vein to the body of rats in every groups.4 After established Model and acupuncture, we use 10% chloral hydrate to anesthesia the rats, tand ake the liver tissue and arterial blood. Liver was immediately make tobe frozen sections to obseve the transmission rate of labeled cells under fluorescence microscope, the blood serum was centrifugated and kept under-70℃for testing.5 Added the blood serum of rats in each group to cell culture medium. We observed the effect of ratss blood serum in each group for MSCs proliferation and differentiation in vitro.Results:1 The bone marrow mesenchymal stem cells we cultured express cell surface antigens CD44, CD71, did not express CD34,which is hematopoietic lineage markers.2 The liver fibrosis rats model we established is proved tobe successful by blood serum markers of liver fibrosis and HE staining.3 The CM-Dil sent a red light excited by red fluorescence after labeled 48 hours.Shape of the cell is clearly visible, the membrane cytoplasm can see fluorescence.4 The liver frozen sections expresse red fluorescent under fluorescent microscope.The normal liver tissue did not express red fluorescent. The fibrosis liver can be seen a little amount of red fluorescence cells.The points acupuncture group and the Connective tissue group expresse can see more red fluorescence cells compared with the liver fibrosis model group. The cells that expresse red fluorescent were round or fusiform shape, which scattered live in the gaps of liver cells.5 Except the normal control group, blood serum of the rest group intervente the bone marrow mesenchymal stem cells training in vitro, the bone marrow mesenchymal stem cells performanced hepatocyte-like shape and expression of albumin was measured.Conclusion:1 MSCs was successfully extracted and cultured, and the rat model of alcoholic liver fibrosis is established in this experiment.2 Red fluorescent CM-Dil successfuliy labed and traced bone marrow mesenchymal stem cells in vivo. 3 Acupuncture points and Connective tissue can promote exogenous bone mesenchymal stem cells tend to migrate to the injured liver in the body of fibrosis rat. Acupuncture treatment of points and the Connective tissue in liver fibrosis in ellular mechanism is likely to mobilize not only undifferentiated bone marrow stem cell in the body's own connective tissue and bone marrow, but also mobilize exogenous bone marrow mesenchymal stem cells, and promote the stem cells differentiate into liver cells to repair injured liver, and ease liver fibrosis. Material base of acupuncture may have a close relationship with the body's bone marrow mesenchymal stem cells and the connective tissue. This prove that the blood serum of liver fibrosis rats can induce bone marrow mesenchymal stem cells into liver cells in vitro; When the cell is cultured in vitro environment, the acupuncture of points and connective tissue can not inprve the ability of liver fibrosis rats' blood serum to induce bone mesenchymal stem cells to differented to live cells in vitro.4 Blood serum in liver fibrosis rats can induced bone marrow mesenchymal stem cells into liver cells in vitro.After hepatic fibrosis model rats were acupunctured points and connective tissue, it's blood serum could not induced more bone marrow mesenchymal stem cells to different into liver cells. This result may be due to that the environment cell cultured in vitro is different with the liver tissue microenvironment in vivo. Although the components of rat blood serum changed, but the blood serum does not replace the microenvironment tha Bone marrow mesenchymal stem cells live in liver tissue in vivo.
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