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研究生:徐曉萍
研究生(外文):Hsu-Hsiao-Ping
論文名稱:高壓氧氣對類紅斑性狼瘡小鼠-紐西蘭黑白鼠雜交第一代小鼠疾病發展之影響
論文名稱(外文):Effects of Hyperbaric Oxygen Therapy of Disease Development of (NZBxNZW)F1 Lupus-like Mice
指導教授:陳紹原
學位類別:碩士
校院名稱:國防醫學院
系所名稱:海底醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:75
中文關鍵詞:高壓氧氣治療全身性紅斑性狼瘡紐西蘭黑白鼠雜交第一代小鼠自體免疫疾病
外文關鍵詞:HBO2 TherapySystemic Lupus ErythematosusSLEautoimmune disease(NZBxNZW)F1
相關次數:
  • 被引用被引用:2
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中文摘要
高壓氧治療(Hyperbaric oxygen therapy)已被廣泛地用於治療或輔助治療許多臨床疾病如減壓症、一氧化碳中毒、空氣栓塞、慢性骨髓炎、燒燙傷等。目前已知高壓氧治療的作用機轉主要為增加組織中氧氣濃度、改變病灶血流和血氧的分佈、增加紅血球的通透性及降低組織水腫等。至於高壓氧對免疫系統的影響,近年來陸續有不同的研究報告提出高壓氧處置可以改變淋巴細胞的組成及分佈,並有降低淋巴細胞增生的現象,其中的機轉雖然尚未明瞭,然而高壓氧對免疫細胞所產生的抑制現象增加了高壓氧處置運用在治療某些自體免疫疾病或器官移植排斥等問題的可行性。
全身性紅斑性狼瘡是一種好發於年輕女性的慢性自體免疫疾病,病人體內會有自發性反應性淋巴球增生的情形,並有大量的自體抗體被製造造成自身組織器官的傷害。本實驗利用與人類全身性紅斑性狼瘡病徵最為相似的動物模式--紐西蘭黑白鼠雜交第一代小鼠(NZB/W F1),在小鼠類紅斑性狼瘡疾病徵狀出現之早期(鼠齡3個月)、中期(鼠齡6個月)與晚期(鼠齡8個月)等時間點,讓小鼠接受連續2星期每天90分鐘,2.5ATA之高壓氧處置,並於各時間點以Albustix半定量試紙檢測老鼠蛋白尿情形,及抽取老鼠之眼窩血,以ELISA檢測老鼠血清中anti-dsDNA autoantibodies IgG titer;以Flow Cytometry檢測老鼠周邊血液及脾臟中淋巴細胞之再分佈情形;並以冷凍切片處理方式,將老鼠腎臟以免疫螢光染色方式檢測老鼠腎臟腎絲球免疫複合物沈積情形與分析老鼠之存活率。結果發現,早期、中期與晚期皆接受高壓氧治療之A組與中期與晚期接受高壓治療之B組,較晚期才接受高壓氧治療之C組與不接受任何高壓氧治療之D組有存活率增加與明顯蛋白尿產生的百分比降低之趨勢;而在血清anti-dsDNA antibodies titer檢測中,A、B兩組亦較C、D兩組為顯著降低;周邊血液之淋巴細胞與脾臟淋巴細胞分佈情形,A、B兩組呈現CD3+(T淋巴細胞)有顯著較C、D兩組降低之情形,而相對地CD19+(B淋巴細胞)則顯著升高。對T淋巴細胞亞群的進一步分析發現A、B兩組之CD4+(輔助型T淋巴細胞)較C、D兩組有顯著降低情形;而CD8+(細胞毒殺性T淋巴細胞)四組動物間並無顯著差異。在腎臟病理組織的免疫螢光染色結果可見A、B兩組免疫複合物沈積於腎絲球的情形,無論在數量上與亮度上皆較C、D兩組少與暗;我們的結果顯示,對疾病發生的早期與中期接受高壓氧處置的老鼠,其疾病發展之過程有被抑制的現象。
Abstract
Hyperbaric oxygen (HBO2) has been widely applied as an adjunctive treatment for a variety of medical disorders, such as carbon monoxide poisoning, decompression sickness, air embolism, chronic osteomyelitis, and burn injuries. The principal known mechanisms of HBO2 are that it increases oxygen concentration in the blood supply, changes the blood flow to the abnormal area, increases the flexibility of red cells, and reduces tissue edema. The effects of HBO2 on the immune system are controversial. Recent studies showed that HBO2 exposure produced changes in the immune system, including lymphocyte redistribution and decreased lymphocyte proliferation. The suppressive function of HBO2 on the immune system might be suggested to be applicable for the treatment of certain autoimmune diseases and allograft rejection problems.
System lupus erythematosus (SLE) is a chronic autoimmune disease that generally affects young women. It is characterized by large numbers of autoreactive lymphocytes reacting to self-antigens. The resulting formation of immune complexes can deposit in tissue and activate the complement cascade. Eventually inflammation causes various forms of tissue damage.
Murine SLE, which provides a good model of human SLE, has been used to study the pathogenesis of this disease. In this study we used the (NZBxNZW) F1 mice, the unique lupus prone hybrid, to evaluate the effects of HBO2 therapy on autoimmune diseases. The mice were divided into 4 groups. 3 groups of mice were treated with HBO2 at 2.5 ATA for 90 min daily over 2 weeks starting from different age of 3-month-old, 6-month-old, 8-month-old, respectively: A (mice given HBO2 protocol at 3-, 6-, and 8-months-old); B (mice given HBO2 protocol at 6- and 8-months-old); C (mice given HBO2 protocol at 8-month-old); D (control group).Our results show that: (1) HBO2 decreases the percentage of positive proteinuria (>2+) in groups A and B and increases their survival rate; (2) HBO2 reduces the anti-dsDNA IgG antibodies titer in groups A and B; (3) HBO2 changes the distribution of lymphocytes in groups A and B (the percentage of CD3+ [T lymphocyte] is decreased, the CD19+ [B lymphocyte] is increased, the subset of T lymphocytes [CD4+] is decreased); (4) groups A and B have less anti-dsDNA IgG immuno-complex deposition and less brightness in the glomerulus.
HBO2 improves the disease expression of (NZBxNZW) F1 in groups A and B mice given the HBO2 protocol at early (3-month-old) and middle (6-month-old) stages of lupus development. Group C mice given the HBO2 protocol at the late stage (8-month-old) of disease development showed no significant beneficial effect.
目錄
頁次
目錄------------------------------------------------------------------------ Ⅰ
表次------------------------------------------------------------------------ Ⅳ
圖次------------------------------------------------------------------------ Ⅴ
中文摘要----------------------------------------------------------------- Ⅶ
英文摘要----------------------------------------------------------------- Ⅸ
第一章 序言------------------------------------------------------------ 1
第一節 高壓氧氣治療-------------------------------------------- 4
壹 高壓氧氣治療之作用機轉---------------------------- 5
貳 高壓氧氣治療之適應症-------------------------------- 6
參 高壓氧與免疫系統--------------------------------------- 6
肆 高壓氧與自體免疫疾病-------------------------------- 8
第二節 全身性紅斑性狼瘡------------------------------------ 10
壹 定義---------------------------------------------------------- 10
貳 流行病學--------------------------------------------------- 11
參 致病因子--------------------------------------------------- 12
肆 病理機轉--------------------------------------------------- 12
伍 SLE的治療與預後------------------------------------- 15
第三節 全身性紅斑性狼瘡之實驗動物模式---------- 16
第二章 實驗目的---------------------------------------------------- 18
第三章 材料與方法------------------------------------------------- 18
第一節 實驗動物------------------------------------------------ 18
第二節 動物高壓艙--------------------------------------------- 19
第三節 實驗動物之分組-------------------------------------- 19
第四節 高壓氧氣處置----------------------------------------- 19
第五節 實驗參數------------------------------------------------ 20
壹 尿液蛋白質濃度測定---------------------------------- 20
貳 自體抗體anti-dsDNA IgG濃度測定------------- 20
參 周邊血液及脾臟淋巴細胞組成分析------------- 22
肆 存活率------------------------------------------------------ 23
伍 腎臟組織病理切片------------------------------------- 23
第六節 數據分析------------------------------------------------ 24
第四章 實驗結果---------------------------------------------------- 25
第一節 高壓氧對存活率之影響--------------------------- 25
第二節 高壓氧對腎臟功能之影響------------------------ 25
第三節 高壓氧對自體抗體anti-dsDNA IgG製造之
影響----------------------------------------------------- 26
第四節 高壓氧對周邊血液與脾臟淋巴細胞分佈之
影響----------------------------------------------------- 27
第五節 高壓氧對腎絲球免疫複合物沈積情形之影
響--------------------------------------------------------- 29
第五章 討論----------------------------------------------------------- 30
第六章 未來方向---------------------------------------------------- 36
第七章 參考資料---------------------------------------------------- 57
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