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GM-CSF in the Wound of Diabetic Rats BCL-2 Expression Levels

Author: ZhangZuo
Tutor: LiZhengWei
School: Dalian Medical University
Course: Surgery
Keywords: Ⅱd iabetic wound healing BCL-2 CD34
CLC: R644
Type: Master's thesis
Year: 2011
Downloads: 4
Quote: 0
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Objective: Wound healing is an evolutionarily conserved, complex multi-cell process. This process involves a variety of cell types, including keratinocytes, fibroblasts, endothelial cells, macrophages, and platelets co-ordination efforts. Diabetes mellitus (diabetes) is genetic factors, immune disorders, microbial infections and toxins, free radical toxins, mental factors and so the role of agents that cause islet dysfunction in the body, caused by insulin resistance and sugar, protein, fat, and a series of water and electrolyte metabolism disorder syndrome. Because the diabetes caused by peripheral nerve lesions, diabetic microangiopathy in patients with difficult to heal wounds pathological basis. Granulocyte macrophage colony stimulating factor (GM - CSF) is a pluripotent growth factor, in the process of wound healing plays an important role. It can induce neutrophil and monocyte / macrophage differentiation, proliferation and activation; improve wound resistance to infection; promote necrotic tissue loss; enhance the proliferation of hematopoietic precursor cells; chemotaxis, activation of endothelial cells, fibroblasts and skin cells, promote their differentiation, proliferation, angiogenesis, granulation tissue formation and epithelial cell migration, proliferation, and further speed up the time; is conducive to promoting healing factor secretion, and reduced wound tissue remodeling [1]. The role of such a wide range of domestic and foreign experts attracted the attention of scholars, they will be GM-CSF used in a variety of wounds, such as cancer, diabetes, skin diseases, surgical wounds and skin ulcers after deep burns, venous ulcer release of tumor chemotherapy and other unhealed wound and speed up the healing. Because in the process of wound healing keratinocytes, fibroblasts, endothelial cells, macrophages, and platelets play an important role and, while GM-CSF plays a catalyst in the meantime, the activation and activation of various cells [2], we believe that GM-CSF in wound healing in diabetes may play a role. Our experimental model of wound healing in diabetic rats the expression of Bcl-2 to simply explore the process of wound healing in diabetic granulocyte-macrophage colony-stimulating factor role.Methods: SPF Level 60 rats, body weight (290±15) g, male or female. Environment in high fat, sugar fed for 4 weeks after modeling, fasting for 12 hours before modeling, can not help but water. STZ prepared with the new 0.1mmol / L citrate buffer PH = 4.2 dilution. Small dose of intraperitoneal injection of STZ 35mg/kg. Blood sugar stable for about 48h-72h. Rats had more than three symptoms, weight gain. Then in the tail vein blood testing., Blood glucose greater than 11mmol / L or more. Check blood sugar and weight standard pathological examination of pancreatic islets of rats for islet cells proved successful model showed no damage. After the modeling randomly selected 45 rats, Rats were anesthetized first shaving the back, making each side of the back midline wound 1.5cm x 1.5cm, full-thickness skin wounds cut to subcutaneous fascia, no bacteria dressing stamped, refueling once daily. 45 randomly selected rats were divided into three groups: A group of diabetes (DM) wound group 15, B group and diabetes + GM-CSF wound group 15 (method after the rats were anesthetized by intraperitoneal injection of GM- CSF), C + thinner wound group of diabetes 15 (method after the rats were anesthetized by intraperitoneal injection of diluent). Rat cages every five minutes. Naked eye after surgery rats 3,5,8,12,18 day wound tissue healing, the rats weighing intraperitoneal anesthesia, blood testing in the tail vein blood glucose levels and record the weight of each rat , fasting blood glucose. Method using a transparent graph paper first 3,5,8,12,18 days measured area of wound healing, wound healing rate also calculated. Along the outer edge of the wound healing beyond the wound tissue was cut about 0.5cm, the producer check the wound after the light microscope view of various types of cells, vascular survival situation. Performed immunohistochemical experiments, observe the wound BCL-2 and CD34 staining by immunohistochemistryResults: Survival of 60 rats, 57, died three cause of death was considered when the wound in the production of local anesthetic overdose. Survival of blood sugar, weight detection, eight complement typeⅡdiabetes meet the criteria, delete the log data. The remaining 49, whichever is 45 included in the study. Naked eye wound healing results: wound 2 days after the formation of the rats were started scab wound. 5 days after wound formation of DM + GM-CSF group compared with a slight change in the other 3 groups, the effect is not obvious. Formed 7 days after the wound from the naked eye DM + GM-CSF group than in the other 3 groups was more significant improvement in wound healing. The healing quality were: DM + GM-CSF group, DM + dilution group, diabetic group. The formation of 8-day wound DM + GM-CSF group of basic wound healing, the other 2 groups slightly different. 18 days after wound formation of DM + GM-CSF group All the wounds healed, the other 2 groups the basic healing, DM group the healing rate difference between the previous two groups. Immunohistochemistry results are: BCL-2: DM + GM-CSF group than in the other two groups was significantly thicker granulation tissue. Granulation tissue from the large number of fibroblasts, collagen fibers and capillaries composition. DM + dilute solution in the cytoplasm of fibroblasts and capillary endothelial cells can be seen a large number of BCL-2 expression. Can be seen in the diabetic group, even the expression of BCL-2 positive. CD34: DM + GM-CSF group showed a large number of granules stained brown, while there are a lot of blood vessels. DM + dilute solution a small amount of brown particles in the visible stain, hair follicle cells have a similar organization exists. Staining was not obvious in the diabetic group, small brown particles.Conclusion:Wounds of diabetic rats in the application of granulocyte-macrophage colony-stimulating factor, may activate, differentiation, proliferation and activation of endothelial cells, fibroblasts and skin cells. BCL-2 expression level (dynamic rules) suggest GM-CSF can inhibit apoptosis, thereby promoting wound healing.

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