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Title: | Artocarpin促進皮膚傷口癒合 Artocarpin Enhances Skin Wound Healing |
Authors: | Chung-Ju Yeh 葉重如 |
Advisor: | 王淑慧(Shu-Huei Wang) |
Keyword: | 傷口癒合,黃酮類化合物,纖維母細胞,角質細胞,內皮細胞,膠原纖維,細胞增生,細胞移動,血管新生, artocarpin,NIH3T3,HaCaT,HUVEC,collagen,cell proliferation,cell migration,angiogenesis, |
Publication Year : | 2015 |
Degree: | 碩士 |
Abstract: | 皮膚可以幫助身體抵抗外來的有害物質和微生物。當皮膚受到傷害時,傷口癒合的過程需要被精密調控以恢復皮膚組織的正常功能。Artocarpin,一種從植物Artocarpus communis中萃取出來的異戊二烯化之黃酮類化合物(flavonoid),在先前的研究中發現有抗發炎和抗腫瘤的效果。本次實驗的目標是要探討artocarpin在傷口癒合中可能的療效和相關的作用機制。
本次實驗中,artocarpin在小鼠的切除傷口模式中加速了傷口癒合。嗜中性球和巨噬細胞的免疫組織化學染色以及小鼠的cytokine array顯示artocarpin經由加速傷口癒合中發炎期的過程來達到調控發炎的效果。在Mallory苯胺藍膠原蛋白染色、天狼星紅苦味酸染色和西方墨點法的實驗結果中,在以artocarpin治療的組別,膠原纖維的堆積有增加的情形。此外,在cytokeratin14 (CK14,角質細胞marker)和proliferating cell nuclear antigen (PCNA)的雙重染色結果顯示下,artocarpin可以經由促進角質細胞的增生來促進表皮再生的過程。在CD31(血管內皮細胞marker)的免疫組織化學染色和西方墨點法實驗結果顯示,以artocarpin治療的組別,有血管新生增加的情形。在細胞實驗中,纖維母細胞、角質細胞和血管內皮細胞在以artocarpin處理後,細胞增生都有顯著的增加,而這些作用可能是藉由extracellular signal–regulated kinase 1/2 (ERK1/2)、P38或Akt所進行調控。在scratch wound assay中,以artocarpin處理纖維母細胞和角質細胞加速了其傷口閉合,以artocarpin處理之角質細胞也展現了增強的細胞移動能力。此外,以artocarpin處理血管內皮細胞可以增進其在matrigel tube formation assay中形成脈管構造的能力。 綜合以上結果,我們的研究顯示artocarpin可以促進傷口癒合,此效果可能是經由加速發炎期的過程的進行、促進膠原纖維堆積、表皮再生和血管新生等過程所造成。這些研究結果顯示了在傷口癒合治療上,artocarpin極具有治療的潛力。 Skin protects the body against harmful substances and microorganisms. When skin is damaged, wound healing needs to be finely regulated to restore the normal function of skin tissue. Artocarpin, a prenylated flavonoid purified from the plant Artocarpus communis, has been reported to have anti-inflammation and anti-cancer properties. The aim of the present study is to evaluate the wound healing potential and the relative therapeutic mechanism of artocarpin. In this study, artocarpin accelerated the wound healing process in a mouse skin excisional wound model. Immunohistochemical staining of neutrophil and macrophage and mouse cytokine array analysis revealed that artocarpin exerts an inflammatory regulation by accelerating the process of inflammatory phase of wound healing. Mallory’s aniline blue collagen stain, picrosirius red stain, and western blot analysis showed increased collagen deposition in the artocarpin-treated group. Furthermore, the results of double staining of cytokeratin 14 (CK-14, a keratinocyte marker) and proliferating cell nuclear antigen (PCNA) showed that artocarpin promote re-epithelialization by enhancing keratinocyte proliferation. Immunohistochemical staining and western blot analysis of CD31 revealed an increase in angiogenesis in the artocarpin-treated group. In the in vitro study, increased proliferation of fibroblasts, keratinocytes, and endothelial cells were observed when treated with artocarpin, and these effects were mediated through activation of extracellular signal–regulated kinase 1/2 (ERK1/2), P38, or Akt. Keratinocytes and fibroblasts treated with artocarpin showed accelerated wound closure in scratch wound assay. Keratinocytes treated with artocarpin also exhibited increased cell migration. Furthermore, artocarpin treatment enhanced the ability of endothelial cells to form tubes in matrigel tube formation assay. Taken together, our studies suggest that artocarpin enhances skin wound healing, possibly through accelerating the inflammatory phase, as well as increasing collagen deposition, re-epithelialization, and angiogenesis. These studies implicate artocarpin as a potential powerful therapeutic agent for the treatment of skin wounds. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52485 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 解剖學暨細胞生物學科所 |
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ntu-104-1.pdf Restricted Access | 3.97 MB | Adobe PDF |
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