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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 紀秀華(Shiou-Hwa Jee) | |
dc.contributor.author | Yu-Ting Huang | en |
dc.contributor.author | 黃鈺婷 | zh_TW |
dc.date.accessioned | 2021-06-08T04:17:05Z | - |
dc.date.copyright | 2010-09-09 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-30 | |
dc.identifier.citation | Al-Kurdi D, Bell-Syer SE and Flemming K. Therapeutic ultrasound for venous leg ulcers. Cochrane Database Syst Rev 2008:CD001180
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22405 | - |
dc.description.abstract | 白斑病灶皮膚的色素回復,需要由位在 ”melanocyte reservoir” 的黑色素母細胞或黑色素細胞開始移動到色素缺失之處。因此,白斑的臨床治療,除了避免並移除會使黑色素細胞破壞的可能原因 (offending factors)之外,主要的目的在於能降低黑色素細胞被破壞 (如: 降低皮膚局部發炎環境之免疫反應),或促進黑色素母細胞或黑色素細胞移動、增生、成熟,以使黑色素回復。若有能達成上述目的的物質或方法,即有潛能可以發展成一新的白斑治療輔助方式,以提高其治癒率。
目前脈衝式超音波已被報告可在各種組織細胞產生的廣泛的生物效應,主要包括:促進纖維母細胞增生及細胞移動力增加、促進成骨母細胞增生及成熟分化、促進纖維母細胞、成骨母細胞及周邊血液單核球細胞分泌血管新生物質IL-8, bFGF, 及VEGF,進而幫助傷口癒合及組織修復等等。而本實驗發現脈衝式超音波的直接刺激會使黑色素母細胞NCCmelb4及黑色素細胞NCCmelan5之移行增加,以1W/cm2 ~ 1.5W/cm2效果較佳。此種刺激黑色素母細胞或黑色素細胞移動增加的效果,使得脈衝式超音波有潛力發展應用為白斑之輔助治療之工具。同時,本實驗顯示,脈衝式超音波的使用在特定能量範圍內,無論直接刺激或是先刺激纖維母細胞NIH 3T3後產生之間接效應,都不會改變黑色素細胞NCCmelan5及黑色素母細胞NCCmelb4之生長曲線。而且,直接刺激時,脈衝式超音波也並不會改變黑色素細胞NCCmelan5的黑色素分泌量。因此,在臨床使用上,若白斑病人接受脈衝式超音波之物理治療,在安全能量範圍下,應不會造成黑色素細胞被破壞產生新病灶。 本實驗中,我們利用cytokine array, Western blot, migration assay等方式證實超音波刺激使NCCmelb4之conditional medium中,分泌有M-CSF,而M-CSF以autocrine方式活化FAK signaling pathway,進而p-FAK表現量增加,進而促進細胞移動增加。 同時本實驗也發現先刺激纖維母細胞NIH 3T3後,產生的間接效應亦會促進黑色素母細胞NCCmelb4或黑色素細胞NCCmelan5移動增加,效果以1W/cm2 ~ 1.5W/cm2較佳。皮膚中,黑色素細胞及黑色素母細胞的生長與移動,受到上皮角質細胞和真皮纖維母細胞的調控。在正常生物組織之中,纖維母細胞的存在與作用,在超音波刺激下,應能有共同加成促進黑色素母細胞或黑色素細胞移動之效應。 白斑的治療過程與成效,常是困難而令人失望的,而且病人也常在治療中遭遇到治療的副作用困擾。如:光療的紅腫、水泡,局部類固醇的皮膚萎縮,口服或局部psoralen的腸胃不適及光敏感刺激……等等,使得病人的醫囑遵從性往往大打折扣。發展一新的有效安全而方便的治療輔助方式,以提高其治癒率,是白斑治療值得努力的方向。而此實驗除了解脈衝式超音波的機械性刺激對黑色素細胞或黑色素母細胞的基礎生物效應之外,亦能做為臨床應用的基礎,證實脈衝式超音波有潛能可以發展成一新的白斑治療輔助方式,以改善目前白斑的治癒率。 | zh_TW |
dc.description.abstract | Background: Repigmentation of depigmented lesions of vitiligo relies on the proliferation and migration of the melanoblasts or melanocytes from melanocyte reservoir to the epidermis of lesional sites. Functional maturation of melanocytes occurs in this process and melanin production recovers the skin tone. Therapy or instrument that can stimulate melanoblasts or melanocytes proliferating or migrating from the melanocyte reservoir may offer another choice of adjuvant therapy for vitiligo to improve efficacy of treatment. Pulsed ultrasound has been reported having stimulatory effects on cell proliferation and migration of fibroblasts and other cells.
Objectives: To clarify the biological effects of ultrasound on melanoblasts and melanocytes, we employ this study to investigate the influences on their proliferation and migration. Methods: NCCmelb4 cells derived from the mouse neural crest have the characteristics of melanocyte precursors (melanoblasts). NCCmelan5 have many of the characteristics of differentiated melanocytes except that they are immortal. Cultured NCCmelb4 and NCCmelan5 were sonicated with pulsed ultrasound. Cell migration and growth were evaluated. In addition, the potentially involved molecular pathways were also determined. Results: Our results show that increased cell migrations are noted in direct stimulation of pulsed ultrasound on melanoblasts (NCCmelb4) and melanocytes (NCCmelan5). Through stimulating on fibroblasts (NIH 3T3), pulsed ultrasound can also indirectly promote cell migration of melanoblasts and melanocytes. Both of the cell growth curves of melanoblasts and melanocytes are not altered by pulsed ultrasound stimulation. Cytokine arrays show that increased secretion of M-CSF in conditional medium of NCCmelb4 and NCCmelan5 by direct pulsed ultrasound stimulation. p-FAK is up-regulated by direct pulsed ultrasound stimulation on melanoblasts. Inhibition of FAK blocks the direct stimulatory effects of pulsed ultrasound on migration of melanoblasts and melanocytes. Combing the results of Western blotting, it’s revealed that M-CSF activates FAK dependent signaling pathway to promote cell migration of melanoblasts. Conclusions: In summary, we have demonstrated that pulsed ultrasound imparts a stimulatory effect on cell migration of melanoblasts and melanocytes via activation of FAK dependent signaling pathway which is induced by M-CSF autocrine secretion. This study not only searches the basic recognition of mechanical stimulation of pulsed ultrasound on melanoblasts and melanocytes, but may also have promising development in the future vitiligo treatment. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T04:17:05Z (GMT). No. of bitstreams: 1 ntu-99-P97421021-1.pdf: 7197577 bytes, checksum: c4ca6c529efe005c31e715f8c21e30fa (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 目錄
一、緒論--------------------------------------------------------------------------------------------------------------1 二、研究目的-------------------------------------------------------------------------------------------------------12 三、研究方法與材料 ---------------------------------------------------------------------------------------------13 (一) 細胞培養---------------------------------------------------------------------------------------13 (二) 超音波刺激---------------------------------------------------------------------------15 (三) 細胞增生分析---活細胞數分析---MTT test ------------------------------------------------------16 (四) 細胞移動分析---wound healing assay -------------------------------------------------------------16 (五) 細胞移動分析--- Boyden chamber migration assay ----------------------------------------------16 (六) 黑色素分泌實驗----------------------------------------------------------------------------------------17 (七) 西方墨點法----------------------------------------------------------------------------------------------18 (八) 細胞激素抗體分析(cytokine array)- ----------------------------------------------------------------19 四、統計分析 -------------------------------------------------------------------------------------------------------19 五、實驗結果 前測 (一)脈衝式超音波會促進HIG-82纖維母細胞株之細胞增生----------------------------------------20 (二)脈衝式超音波會促進HIG-82纖維母細胞株之細胞移動增加----------------------------------20 (三)脈衝式超音波會促進NIH 3T3纖維母細胞株之細胞移動增加-------------------------------21 第一部份——脈衝式超音波對黑色素母細胞及黑色素細胞之直接效應------------------------------22 (一) 脈衝式超音波直接刺激不會改變黑色素母細胞株NCCmelb4之細胞生長曲線----------22 (二)脈衝式超音波直接刺激不會改變黑色素細胞株NCCmelan5之細胞生長曲線------------22 (三)脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之細胞移動增加--------------22 (四)脈衝式超音波直接刺激會促進NCCmelan5黑色素細胞株之細胞移動增加----------------23 (五)脈衝式超音波直接刺激不會改變NCCmelan5黑色素細胞株之黑色素分泌量------------23 第二部份——脈衝式超音波對黑色素母細胞及黑色素細胞之間接效應-----------------------------24 (一)脈衝式超音波間接刺激不會改變黑色素母細胞株NCCmelb4之細胞生長曲線-----------24 (二)脈衝式超音波間接刺激不會改變黑色素細胞株NCCmelan5之細胞生長曲線-------------24 (三)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加 (wound healing assay)---- -------------------------------------------------------------25 (四)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加 (wound healing assay)--------- --------------------------------------------------------25 (五)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加 (Boyden chamber assay)-------- -----------------------------------------------------26 (六 ) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加 (Boyden chamber assay)--------------------------------------------------------------26 第三部份——脈衝式超音波對黑色素母細胞及黑色素細胞生物效應之分生機制---------------27 (一) 脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之p-FAK表現在超音波刺激 後第24小時增加--------------------------------------------------------------------------------------27 (二) 脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之p-FAK表現增加,而此現 象會被FAK inhibitor 14 (sc-203950)抑制-----------------------------------------------------27 (三) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelb4之細胞移動增加,但此效應會 被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay)------------------28 (四) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelan5之細胞移動增加,但此效應 會被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay)------------------28 (五) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelb4之conditional medium中之 M-CSF分泌增加----------------------------------------------------------------------------------------29 (六) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelan5之conditional medium中之 M-CSF分泌增加--------------------------------------------------------------------------------------29 (七) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加,但此效應會被M-CSF antibbody 抑制 (wound healing assay)------------29 (八) 脈衝式超音波直接刺激黑色素母細胞株NCCmelb4後之conditional medium會促進 NCCmelb4之p-FAK表現增加,但此效應會被M-CSF antibody 抑制--------------------30 (九) 脈衝式超音波直接刺激會促進纖維母細胞株NIH 3T3之conditional medium中之多種 chemokine分泌增加-------------------------------------------------------------------31 六、討論------------------------------------------------------------------------------------------------------------31 七、參考文獻------------------------------------------------------------------------------------------------------37 八、圖表 (圖一) 超音波刺激系統-----------------------------------------------------------------------------------42 (圖二)脈衝式超音波會促進HIG-82纖維母細胞株之細胞增生-----------------------------------43 (圖三)脈衝式超音波會促進HIG-82纖維母細胞株之細胞移動增加-----------------------------44 (圖四)脈衝式超音波會促進NIH 3T3纖維母細胞株之細胞移動增加--------------------------45 (圖五)脈衝式超音波直接刺激不會改變黑色素母細胞株NCCmelb4之細胞生長曲線----46 (圖六)脈衝式超音波直接刺激不會改變黑色素細胞株NCCmelan5之細胞生長曲線------47 (圖七)脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之細胞移動增加--------48 (圖八)脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之細胞移動增加--------49 (圖九)脈衝式超音波直接刺激會促進NCCmelan5黑色素細胞株之細胞移動增加----------50 (圖十)脈衝式超音波直接刺激會促進NCCmelan5黑色素細胞株之細胞移動增加----------51 (圖十一)脈衝式超音波直接刺激不會改變NCCmelan5黑色素細胞株之黑色素分泌量----52 (圖十二)脈衝式超音波間接刺激不會改變黑色素母細胞株NCCmelb4之細胞生長曲線---53 (圖十三)脈衝式超音波間接刺激不會改變黑色素細胞株NCCmelan5之細胞生長曲線-----54 (圖十四)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加(wound healing assay)----------------------------------------------------------55 (圖十五)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加(wound healing assay)---------------------------------------------------------56 (圖十六)脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加(wound healing assay)-------------------------------------------------------57 (圖十七) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加(wound healing assay)------------------------------------------------------58 (圖十八) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加 (Boyden chamber assay)-----------------------------------------------59 (圖十九) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加 (Boyden chamber assay)------------------------------------------------60 (圖二十) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加 (Boyden chamber assay)----------------------------------------------61 (圖二十一) 脈衝式超音波間接刺激會促進黑色素母細胞株NCCmelan5之 細胞移動增加 (Boyden chamber assay)-------------------------------------------------62 (圖二十二) 脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之 p-FAK表現在超音波刺激後第24小時增加--------------------------------------------63 (圖二十三) 脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞株之 p-FAK表現增加,而此現象會被FAK inhibitor 14 (sc-203950)抑制-------------------64 (圖二十四) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelb4之細胞移動增加, 但此效應會被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay)-------------65 (圖二十五) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelb4之細胞移動增加, 但此效應會被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay) ------66 (圖二十六) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelan5之細胞移動增加, 但此效應會被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay) -----------67 (圖二十七) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelan5之細胞移動增加, 但此效應會被FAK inhibitor 14 (sc-203950) 抑制(wound healing assay) -----------68 (圖二十八) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelb4之 conditional medium中之M-CSF分泌增加----------------------------------------------69 (圖二十九) 脈衝式超音波直接刺激會促進黑色素細胞株NCCmelan5之 conditional medium中之M-CSF分泌增加---------------------------------------------70 (圖三十) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加,但此效應會被M-CSF antibbody 抑制(wound healing assay)----71 (圖三十一) 脈衝式超音波直接刺激會促進黑色素母細胞株NCCmelb4之 細胞移動增加,但此效應會被M-CSF antibbody 抑制(wound healing assay)----72 (圖三十二) 脈衝式超音波直接刺激黑色素母細胞株NCCmelb4後之conditional medium會促進 NCCmelb4之p-FAK表現增加,但此效應會被M-CSF antibody 抑制-----------------73 (圖三十三) 脈衝式超音波直接刺激會促進纖維母細胞株NIH 3T3之 conditional medium中之多種chemokine分泌增加---------------------------------------74 (圖三十四) 脈衝式超音波直接刺激會促進NCCmelb4黑色素母細胞之 M-CSF分泌以autocrine的方式活化FAK dependent signaling pathway--------------75 | |
dc.language.iso | zh-TW | |
dc.title | 脈衝式超音波對黑色素母細胞及黑色素細胞之生物效應 | zh_TW |
dc.title | Biological effect of pulsed ultrasound on melanoblast and melanocyte | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 陳文翔(Wen-Shiang Chen) | |
dc.contributor.oralexamcommittee | 周祖述(Tzuu-Shuh Jou) | |
dc.subject.keyword | 脈衝式超音波,黑色素母細胞,細胞移動,M-CSF,FAK, | zh_TW |
dc.subject.keyword | pulsed ultrasound,melanoblast,migration,M-CSF,FAK, | en |
dc.relation.page | 75 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2010-07-30 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
顯示於系所單位: | 臨床醫學研究所 |
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