系統識別號 U0026-1602201710440200
論文名稱(中文) 探討在新生兒腦缺氧動物模型中流體剪力及紅黴素刺激之內皮細胞作用
論文名稱(英文) Shear stress and erythromycin stimulations in endothelial cells for treatment of neonatal hypoxic-ischemia brain injury
校院名稱 成功大學
系所名稱(中) 細胞生物與解剖學研究所
系所名稱(英) Institute of Cell Biology and Anatomy
學年度 105
學期 1
出版年 106
研究生(中文) 江宜倫
研究生(英文) Yi-Lun Chiang
學號 t96974032
學位類別 碩士
語文別 英文
論文頁數 40頁
口試委員 指導教授-吳佳慶
中文關鍵字 內皮細胞分化  流體剪力  腦缺氧模型  紅黴素  CXCR4 
英文關鍵字 endothelial differentiation  shear stress  brain hypoxic-ischemia model  erythromycin  CXCR4 
中文摘要 幹細胞可分化成多種細胞,其中亦包含血管內皮細胞,然而,諸多因素導致幹細胞取得不易且分化效果不彰。本實驗室先前研究發現,可利用流體剪力模擬血流,結合化學因子,刺激幹細胞分化形成內皮細胞,然其分子調控機制尚不明。多項研究指出在流體剪力刺激下,會活化幹細胞的生長分化,調控動靜脈內皮細胞分化。
本研究將以人類臍靜脈內皮細胞(human umbilical vein endothelial call, HUVEC)建立腦缺氧動物模型,以利流體剪力刺激分化後之細胞應用。在腦缺氧動物模型中,給予臍靜脈內皮細胞,將降低腦損傷,並減少大腦細胞死亡。層流(laminar)刺激後之臍靜脈內皮細胞較未刺激之細胞無明顯修復效果。紅黴素(erythromycin, EM)處理後之臍靜脈內皮細胞具有較高CXCR4表現。在不同層流及紅黴素結合方式刺激下,層流刺激後再以紅黴素處理之細胞具有較多CXCR4表現,並有高度修復大腦之效果。在此動物模型中,我們成功建立內皮細胞修復大腦組織模型;並可藉由增加CXCR4表現,而增加內皮細胞修復效用。
英文摘要 Stem cells can differentiate to endothelial lineage as a source for tissue engineering of vessels. However, the rarity of cell source and low differentiation efficiency are the main limitation for stem cell therapy in vessel regeneration. Our previously study demonstrated a successful endothelial cell (EC) differentiation by integration of biomechanical force and chemical factors, but the mechanism is still unknown. Furthermore, we build up a cell therapy of brain hypoxic-ischemia (HI) model by using human umbilical vein endothelial cells (HUVECs). The differentiated cells will apply to this animal model in the future. By static HUVECs transplantation in brain HI model, brain injury and cell death were decreased. Laminar shear stress (LSS)-treated HUVECs group in animal model had not significantly repair function than static HUVECs. Previously research indicates that CXCR4, a chemoattractant receptor, is reduced by LSS. One paper showed that erythromycin (EM) increased CXCR4 expression. Therefore, we combined LSS with EM to stimulate HUVECs. We found that EM treatment after LSS stimulation of HUVEC transplantation was useful for rescuing brain injury. In this study, we built up successful cell therapy for brain injury model. Furthermore, repair function of ECs can be enlarged by increasing EM-induced CXCR4 expression.
論文目次 中文摘要 i
Abstract ii
Acknowledgement iii
Contents iv
Background 1
Stem cell in vascular therapy 1
Embryonic stem cells 1
Adult stem cells 1
Endothelial progenitor cells 2
Shear stress in vascular system 3
Mechanotransduction of stem cell differentiation and proliferation 5
Animal model of cell therapy in neonatal hypoxic-ischemia (HI) brain injury 6
Hypothesis 8
Specific aims 8
Significance 9
Materials and methods 10
Cell isolation and culture 10
Animal model 10
Shear stress experiments 11
Western Blot 11
Reverse transcription-polymerase chain reaction (RT-PCR) 12
Cell death assay 12
Immunohistological staining 12
Nissl staining 13
Terminal dUTP Nick-End Labeling (TUNEL) Staining 13
In vivo cell tracing model 13
Cell migration assay 13
Statistics 14
Result 15
Erythromycin combine with LSS provide beneficial effects for HUVEC 15
Decrease of CXCR4 by siRNA reduced expression of CXCR4 16
Application of HUVECs as EPC source for neonatal HI brain injury model 17
Discussion 19
References 22
Figures 27
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