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標題: | 植入臍帶間質衍生之內皮前驅細胞可藉由抑制細胞凋亡及發炎反應防止腎臟缺血再灌流之傷害 Endothelial progenitor cells from Wharton’s Jelly prevent kidney ischemia/reperfusion injury by inhibiting apoptosis and inflammation |
作者: | Fu-Bin Chang 張富斌 |
指導教授: | 陳玉怜(Yuh-Lien Chen) |
關鍵字: | 急性腎損傷,細胞凋亡,發炎反應,活性氧自由基,內皮前驅細胞, acute kidney injury,apoptosis,inflammation,reactive oxygen species,EPC, |
出版年 : | 2012 |
學位: | 碩士 |
摘要: | 急性腎損傷是個高致病率及高死亡率的疾病,在臨床上最常見的原因是缺血性缺氧及再灌流傷害(I/R injury)所造成的,但是目前還沒有有效的治療藥物,因此,能發展出新興的治療方式是眾所期盼的。至今已經有許多證據指出,內皮前驅細胞(EPC)具有改善缺血性缺氧的組織傷害,由於目前EPC的主要來源是從骨髓與周邊血液分離而來,其EPC的數量會受到許多因素影響。在本篇研究中,利用小鼠建立腎臟I/R injury的動物模式,並探討由臍帶間質(Wharton’s jelly)所分離的EPC是否具有治療潛力。首先由Wharton’s jelly分離培養出Mesenchymal cells,使其分化成EPC,以吞噬乙醯化的低密度脂蛋白(acetylated-LDL)能力及表現內皮細胞特徵做鑑定。為了方便觀察EPC在組織中的位置,我們事先由Q-tracker標記EPC後再植入到腎臟的包囊中。根據實驗結果,經過I/R injury之後,血尿氮素(BUN)及creatinine會顯著上升,相較之下,植入EPC後不只其表現量有下降情形,利用蘇木素-伊紅(H&E)染色對組織傷害的評分也有下降的結果。此外,透過TUNEL檢測和H&E染色可以發現植入EPC後也可以有效抑制血管內皮細胞、腎絲球及腎小管細胞的凋亡,除了細胞凋亡之外,也可以降低I/R injury之後ROS (reactive oxygen species)的生成量。另外,我們也利用免疫組織化學染色,觀察EPC植入組其發炎因子Keratinocyte-derived Cytokine (KC)的表現量會降低。在微血管密度方面,EPC也可以恢復I/R injury後的微血管密度,並且發現其具有嵌入微血管內皮情形,最後利用纖維母細胞的marker S100A4進行免疫組織化學染色,結果顯示,雖然S100A4表現的細胞數量會隨術後時間增加而變多,但EPC植入之後表現S100A4的細胞數量減少及減緩內皮間質細胞轉型(endoMT)的現象。綜合以上實驗結果,我們認為由Wharton’s jelly所分離出的EPC對於急性腎損傷的患者是個很有發展潛力的治療方式,可能藉由促進血管新生與抑制細胞凋亡、ROS、發炎反應及減緩腎臟纖維化潛力做為改善急性腎損傷的治療方式。 Ischemia/reperfusion (I/R) injury in the kidney is a major cause of acute renal failure in humans and is associated with significantly high mortality. Therefore, to develop a new therapeutic strategy is highly desirable. Accumulating evidence has indicated that endothelial progenitor cells (EPCs) improved the function of ischemic tissues. However, the number and the quality of EPCs from bone marrow and peripheral blood to treat ischemic tissues are limited. In this study, we examined the therapeutic potential of the implantation of EPCs isolated from Wharton’s jelly on renal I/R injury in mice. Mesenchymal cells from Wharton’s jelly to differentiate into EPCs demonstrated by the incorporation of acetylated low-density lipoprotein and expression of the endothelial cell-specific markers. To visualize the localization of transplanted EPCs, cells were labeled with Q-tracker and then were injected into renal capsule. Mice with renal I/R treatment significantly showed the increased concentrations of blood urea nitrogen and creatinine. In contrast, EPC transplantation decreased the levels. The renal injury score induced by I/R injury was significantly decreased by EPCs transplantation. In addition, transplantation of EPCs effectively inhibited I/R-induced cell apoptosis in endothelial cells, glomerulus, and renal tubular cells, as demonstrated by TUNEL assay and hematoxylin-eosin staining. The transplantation of EPCs also significantly reduced reactive oxygen species production in response to I/R injury. The expression of Keratinocyte-derived Cytokine (KC), an inflammatory chemokine, was decreased by EPCs transplantation via immunostaining. Furthermore, EPC transplantation maintains the microvascular density and incorporates into capillary in response to I/R injury. Moreover, EPC effectively reduced I/R-induced endothelial mesemchymal transition (endoMT), as demonstrated by immunostaining with S100A4, a fibroblast marker. Taken together, this study strongly suggested that transplantation of EPCs from Wharton’s jelly from umbilical cord may provide as a novel therapy for ischemic acute renal injury by promoting angiogenesis and inhibiting apoptosis, inflammation as well as kidney fibrosis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65268 |
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顯示於系所單位: | 解剖學暨細胞生物學科所 |
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