Dissertation > Excellent graduate degree dissertation topics show

Empirical Study of Bone Marrow-derived Endothelial Progenitor Cells in Treatment of Diabetic Rats with Hind Limb Ischemia

Author: ZhouChenGuang
Tutor: MengJianBo
School: Hebei Medical University
Course: Internal Medicine
Keywords: Endothelial progenitor cells Bone marrow mononuclear cells Diabetes Lower limb ischemia Angiogenesis Vascular endothelial growth factor Vascular density
CLC: R587.1
Type: Master's thesis
Year: 2010
Downloads: 80
Quote: 0
Read: Download Dissertation

Abstract

Background: As we all know, diabetes mellitus (DM) is a common threat to human life and health and disease. Arterial occlusion caused by lower limb ischemia is one of the four vascular complications of DM, clinical treatment is difficult, amputation rate, accounting for 50% of non-traumatic amputation. As stem cell transplantation technology continues to evolve, people have begun to apply this technique to the study of diabetic lower limb ischemia. Endothelial progenitor cells (endothelial progenitor cells, EPCs) as early as 1997 in the peripheral blood, and proved to play an important role in postnatal angiogenesis (angiogenesis). This study attempts to by EPCs transplantation in the treatment of diabetic lower limb ischemia in animal experiments, and its further clinical application reasonably lay a preliminary theoretical foundation. The first part of endothelial progenitor cells in the bone marrow mononuclear cell transplantation in the treatment of diabetic lower extremity ischemia Objective: diabetic rat bone marrow density gradient centrifugation from mononuclear cells (mononuclear cells, MNCs) to diabetic rats limb ischemia parts local injection of cultured from the same number of MNCs obtained after the injection of EPCs angiogenic effect, and the same amount of bone marrow MNCs and the role of angiogenesis in EPCs compared, in which the role of bone marrow stem cell transplantation and EPCs . Methods: 98 healthy Wistar rats, 10g / L of streptozotocin (STZ) solution 65mg/kg body weight by intraperitoneal injection to achieve the standard of diabetes, 56 ligation of the left femoral artery and main branches of production diabetic lower limb ischemia were randomly divided into A, B, C, D four groups (n = 14). MNCs application density gradient centrifugation method is obtained from the bone marrow fluid extracted by the MNCs, EPCs are inoculated with 10? Cultured in alpha-MEM medium S and growth promoting cytokines 7 obtained a yield of approximately 10%. Transplantation 10 sites for the collection of the cells in PBS suspension uniform injection of the experimental rats in ischemic limbs. A group of transplanted cells 4 × 106 MNCs; B group transplanted cells from an equal number of MNCs with group A cells cultured in vitro, the EPCs (number of about 4 × 105); C group transplanted cells and B group EPCs cells equal to the number of MNCs; D group was injected with the same amount of liquid in PBS. 7 days after the injection, each group of 6 rats were surgical site the muscle derived weighing homogenized, ELISA method measuring VEGF levels. 28 days after the injection, the rats derived immunohistochemistry count density of blood vessels. Results: A group, B group, C group significantly higher than those of Group D in the local VEGF levels and vascular density on the two indicators, statistical significance; regardless of VEGF levels in group A and group B between the two groups or vascular density were not significant differences; comparison between group B and group C, group B content of VEGF and microvessel density two indicators were higher than in group C, results were significantly different. Conclusion: a local injection of bone marrow stem cells can promote angiogenesis in ischemic limbs of diabetic rats. 2 local injection of EPCs amount equal to the amount of MNCs case the ischemic limbs angiogenesis after local injection of EPCs is superior to the local injection of MNCs. 3 bone marrow MNCs directly injected into diabetic lower limb ischemia locally with this number of MNCs culture purified endothelial progenitor cells after local injection, both in promoting angiogenesis was no significant difference said Mingneipizu cells in the bone marrow mononuclear play a leading role in angiogenesis after cell transplantation. The second part of diabetes on bone marrow-derived endothelial progenitor cell transplantation angiogenic role in the lower limb ischemia Objective: To compare receptor (both diabetic lower extremity ischemia in rats or are non-diabetic lower limb ischemia in rats) next, whether the diabetic rats and normal rat bone marrow EPCs after local injection of angiogenesis. Whether the same graft (both diabetic rat bone marrow EPCs were normal rat bone marrow EPCs) injection case, the diabetic the limb ischemia rats and non-diabetic lower limb ischemia in rats angiogenesis. Methods: 24 diabetic rats with the same number of normal rats, bone marrow MNCs and were EPCs training, the other from each of 36 rats The diabetic lower limb ischemia model and non-diabetic lower limb ischemia model, to be modeling successful and stable after 1 week, each model group were randomly divided into diabetic lower limb ischemia group: A1, B1, C1, each group 12; non-diabetic lower limb ischemia group: A2, B2, C2, 12 in each group. The diabetic rat EPCs cultured for 7 days after injection A1 and A2 group left lower limb ischemia in rats local, take the same number of training days in normal rats EPCs B1 and B2 group left lower limb ischemia in rats injected in the same number Partly, C1 and C2 of the left lower limb ischemia injected locally in the same manner the same amount of PBS solution. 7 days after injection of 6 rats from each group were drawn measured VEGF levels. Drawn after 28 days, immunohistochemical staining count left lower limb microvessel density of rats in each group. Results: the same receptor, the graft groups were compared: diabetic lower limb ischemia groups VEGF levels and microvessel density, P values ??were lt; 0.05, resulting in significant differences in group A1 and B1 group of local VEGF levels and vascular density increased significant difference compared with the C1 group, but there was no significant difference between group A1 and B1 group between the two groups. Non-diabetic lower limb ischemia VEGF levels and microvessel density, P values ??were lt; 0.05, resulting in significant difference between group A2 and B2 local VEGF levels and vascular density increased than those in the C2 group, the difference was significant, but there was no significant difference in the group A2 and B2 group between the two groups. Graft, each recipient group comparison: the same transplant bone marrow EPCs or PBS solution, A2, B2 and C2 group (non-diabetic lower limb ischemia group) both local VEGF levels and vascular density as compared with the corresponding A1, B1 C1 group (diabetic lower limb ischemia group) increased, P values ??were lt; 0.05, resulting in a statistically significant difference. Conclusion: a local injection of diabetic rat bone marrow progenitor cells and injection of normal rat endothelial progenitor cells were significantly promote angiogenesis, but no significant difference between diabetic transplant \The cells had no significant effect. Injection of the same bone marrow-derived endothelial progenitor cells, angiogenesis of non-diabetic rats limb ischemia compared to diabetic rats significantly, the diabetic transplant \

Related Dissertations

CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Islet disease > Diabetes
© 2012 www.DissertationTopic.Net Mobile