博碩士論文 104821604 詳細資訊




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姓名 德薇(Apriska Dewi Sipayung)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 共生菌痤瘡丙酸桿菌之葡萄糖發酵在小鼠中磷酸鈣誘導之搔癢以及促發炎介白素-6的產生上的效果。
(Effect of Glucose Fermentation of Commensal Propionibacterium acnes Bacteria on the Calcium Phosphate-Induced Pruritus and the Production of Pro-inflammatory IL-6 in Mice.)
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摘要(中) 晚期腎臟病人經過腹膜透析以及血液透析常被搔癢的症狀給困擾著,此症狀也被稱為尿毒症瘙癢。尿毒症瘙癢被發現與發炎細胞激素、鈣離子、磷酸鹽升高有關。鈣離子及磷酸鹽代謝不正常導致磷酸鈣在皮膚裡形成,並刺激搔癢受器。在長期搔癢的症狀中,在皮膚內神經可能會有次級的改變,以及中樞神經系統可能會僧強隊搔癢的感知。研究證實,背根神經節(DRG)在周邊及中樞感知處理以及發炎進程以及神經性疼痛藉由活化ERK扮演了重要的角色。痤瘡丙酸桿菌為皮膚上的共生菌,在小鼠皮膚中有抗發炎的功能並且被發現能調控不同種類的促發炎細胞激素/趨化因子。在小鼠注射個劑量的磷酸鈣後出現劇烈的搔抓以及IL-6呈現劑量依賴性的上調。體外(in vitro)的實驗結果顯示IL-6的活化促進調控pERK以及pBTK的表現。我們發現痤瘡丙酸桿菌的發酵產物能緩解搔抓以及抑制調控IL-6表現。針對痤瘡丙酸桿菌表面蛋白,唾液酸酶的免疫染色確認,在皮膚真皮層有痤瘡丙酸桿菌移生。因此,抗發炎之痤瘡丙酸桿菌可以成為透析病人IL-6、pERK、pBTK造成的尿毒搔癢症好的治療性應用。
摘要(英) Pruritus is a common and bothersome symptom in patients undergoing peritoneal and hemodialysis with end-stage renal disease also called uremic pruritus. Uremic pruritus is found to be associated with elevated level of inflammatory cytokines, calcium and phosphate. The abnormalities of calcium and phosphate metabolism may lead to the formation of calcium phosphate (CaP) crystals in the skin, which may stimulate itch receptors. During prolonged pruritus, there may be secondary changes occur in nerves in the skin and perhaps the central nervous system which enhances the perception/sensation of itch (central sensitization). Research evidenced that, dorsal root ganglion (DRG) has an integral role in the modulation of peripheral and central sensory processing and development of inflammation and neuropathic pain via ERK activation. Propionibacterium acnes (P. acnes) a skin commensal bacterium, can function as an anti-inflammatory in the mice skin and found to regulate the function of different pro-inflammatory cytokines/ chemokines. Injection of two different doses of CaP in mice showed vigorous scratching and IL-6 up-regulation in dose dependent manner. In vitro results showed that pERK and pBTK get up regulated upon IL-6 activation. The fermentation product of P. acnes was found to relieve from scratching and down-regulated the IL-6 level. Immunostaining against P. acnes surface protein, sialidase confirmed the colonization of P. acnes in the dermal layer of skin. So, anti- inflammatory of P. acnes can be a good therapeutic approach against IL-6, pERK, and pBTK, for the treatment of uremic pruritus in dialysis patients.
關鍵字(中) ★ 尿毒搔癢症
★ 痤瘡丙酸桿菌
★ 促發炎
★ 介白素-6
★ 發酵
關鍵字(英) ★ Uremic pruritus
★ P. acnes
★ Pro -inflammatory
★ IL-6
★ Fermentation
論文目次
摘要……………………………………………………………………………….……………….i
Abstract…………………………………………………………………………………………...ii
Acknowledgements…………………………………………………...………………………….iii
Table of Contents…………………………………………………………………………………iv
List of Figures……………………………………………………………………………………vi

I. Introductions
1. Skin microbiome………………………………………………………………………1
1.1 Propionibacterium acnes………………………………………………………….......1
2. Fermentation of Propionibacterium acnes…………………………………………….2
3. Calcium Phosphate, Cytokine and Inflammation…………………...…………………3
4. Skin Dialysis Patients………………………………………………………………….4
5. Spectrophotometer Measurement Principal…………………………………………...5
6. Principle of Elisa………………………………………………………………………5
7. Immunofluorescence Principal………………………………………………………...6
II. Materials
1. Apparatus of Instruments……………………………………………………………...7
2. Reagents……………………………………………………………………………….7
III. Methods………………………………………………………………………………….11
1. Growth of Bacteria…………………………………………………………………...11
2. Prepared the Chemicals……………………………………………………………....11
3. Prepared the Sample………………………………………………………………….12
4. Measure Skin Protein Concentration…………………………………………………12
5. ELISA………………………………………………………………………………..12
6. Immunofluoroscence…………………………………………………………………12
7. Immunofluorescence IL-6 Itchy, Non- itchy Skin Biopsies Disease…………………13
8. Western Blot………………………………………………………………………….14
8.1 Prepared the Plates……………………………………………………...…………….14
8.2 Isolates DRG………………………………………………………...……………….14
8.3 Making Gel…………………………………………...………………………………15
8.4 Running Gel, Transfer and Blocking…...…………………………………………….15
8.5 Added Antibody...……………………………………………………………………15
8.6 GAPDH………...…………………………………………………………………….16
IV. Results
1. Measurement of spontaneous scratching behavior………………………………...…17
2. Effect of CaP deposition on itching and IL-6 regulation……….…………………….17
3. Effect on glucose fermentation on CaP induced pruritus…………………………......17
4. Role of BTK in CaP induced pruritus……………..………………………………….18
5. P.acnes colonization glucose fermentation………………...………………………...18
6. In- vitro data showing IL-6 induced pbtk and perk expression with and without……..19
ibrutinib
7. Itchy and non- itchy skin biopsies patients…………………………………………...19
V. Discussions………………………………………………………………………………20
VI. References……………………………………………………………………………….22
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指導教授 黃俊銘(Chun- Ming Huang Eric Huang) 審核日期 2017-7-26
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