臺灣博碩士論文加值系統

烏腳病是一種肢體末端血管的慢性疾病，主要的症狀包括肢體顏色變黑、潰瘍與肢體末端發生壞疽等，主要發生於台灣的西南部一帶，而腐植酸 (humic acid) 被認為是造成烏腳病的原因之一。在當地的井水中發現腐植酸含量高達 200 ppm，當地居民平均每日攝食腐植酸 400 mg。以放射性同位素追蹤動物食用腐植酸後之殘留結果，發現經過 24 小時之後，動物體內仍會有約 60% 的腐植酸殘留，將自烏腳病盛行區域的井水中純化出之腐植酸注入小鼠中，則會有相似烏腳病的臨床症狀發生，如尾巴與腳的顏色變黑等。由於烏腳病患都屬於家庭經濟較差、飲食營養較缺乏之人家，且到目前為止，以飲食方式攝食腐植酸，導致烏腳病產生之動物模式尚未出現，因此本研究於第一部分配合營養因子施以 Wistar 雄性大白鼠高濃度 (1,000 ppm) 的腐植酸，取其血液進行分析。分析結果顯示，將腐植酸加入飼料中餵食大鼠八週之後，腐植酸對大鼠之毒作用並不明顯，但飲食中缺乏蛋白質會造成血液中紅血球、血紅素及血比容數值下降，以及血漿中三酸甘油酯、白蛋白及尿素氮含量下降，此一結果與烏腳病患的血液分析結果相似，顯示烏腳病患其血液生化指標的改變，可能主要是飲食中蛋白質不足，以致營養缺乏，再加上攝食腐植酸所導致的；除此之外，許多研究亦證實腐植酸對人類紅血球細胞會造成氧化壓力等傷害，因此在第二部分的實驗中取腐植酸 2,000 ppm，以灌食或腹腔注射的方式，配合不同酪蛋白含量的飼料，餵養大鼠四週之後犧牲，分析其紅血球、血漿及肝臟細胞內抗氧化物質與脂質過氧化之表現，結果顯示，腐植酸會對細胞造成氧化壓力，且若生物體蛋白質攝取量不足，則會加劇腐植酸之影響，對生物體造成較嚴重之傷害，以腹腔注射處理之大鼠其表現與相同條件下以灌食方式之大鼠無異；另外取大鼠的紅血球加入具有溶血活性之皂素 (0.05‰ 與 0.1‰)，反應結果顯示，攝食 20% 酪蛋白之大鼠其紅血球幾乎不受皂素影響，但若攝食 10% 或 0% 酪蛋白之大鼠，與皂素反應後其紅血球溶解情形均增加，顯示其對皂素之耐受性較差，然而腐植酸對紅血球之耐受性則沒有明顯影響。顯示飲食中蛋白質缺乏可能使紅血球變為較脆弱，一旦腐植酸存在時，則會提高氧化壓力，使紅血球受損而出現溶血。由實驗結果推測，烏腳病的真正致病因子，可能是蛋白質缺乏，導致紅血球脆弱易受腐植酸等氧化傷害而形成烏腳病。

Blackfoot disease is a chronic, peripheral vascular occlusive disease and prevalent in southern Taiwan. The major symptoms include discoloration, ulceration, and gangrenous of the extremities. Humic acid is considered to be one of the pathogens of blackfoot disease. The researchers treated mice using intra-abdominal injection with humic acid, and the symptoms which like blackfoot disease appeared. However, the animal model of that humic acid induces blackfoot disease by food and drink has not been yet built. Furthermore, most of the patients with blackfoot disease are poor and undernourished. In the first, part of this study the relationships between nutritional factor and blackfoot disease and the toxic expression of humic acid after ingestion were investigated. The rats were fed the diet with different content of casein (20%, 10%, 0%) and humic acid (1,000 ppm), then the blood of rats were analyzed.
The results showed that the level of red blood cells, hemoglobin, hematocrit, triglyceride, albumin and blood urea nitrogen were decreased when the content of casein was scanty, these phenomenons were similar to those of blackfoot disease patients, but humic acid did not significantly affect the body weight and biochemical index of rats.
Many reports indicated that humic acid cause the oxidative stress and injure red blood cells of human. To investigate the effects of oxidative stress when humic acid was ingested and the relationship between oxidative stress and nutritional factor, the rats were fed the diet with different content of casein (20%, 10%, 0%) and treated with humic acid by oral administration and intraperitoneal injection, raspectively. Then the blood and liver of rats were analyzed. The results showed that the activity of catalase and SOD and the ratio of GSH/GSSG decreased, and the levels of TBARS in liver and plasma increased when humic acid was present, and lacking protein may make it more serious and make red blood cells fragile. These results indicated that lacking protein seems to be the major etiologic factor of blackfoot disease, and humic acid provides the oxidative stress to induce blackfoot disease.

中文摘要---------------------------------------------------1
英文摘要---------------------------------------------------3
壹、 文獻整理
一、烏腳病 (blackfoot disease）----------------------------6
二、砷與烏腳病之關係---------------------------------------9
三、腐植酸與烏腳病之關係----------------------------------11
貳、酪蛋白對腐植酸於大鼠毒作用之影響
一、前言--------------------------------------------------17
二、材料與方法--------------------------------------------19
三、結果--------------------------------------------------26
四、討論--------------------------------------------------29
圖表------------------------------------------------------31
參、酪蛋白對腐植酸於大鼠氧化傷害之影響
一、前言--------------------------------------------------44
二、材料與方法--------------------------------------------45
三、結果--------------------------------------------------52
四、討論--------------------------------------------------56
圖表------------------------------------------------------59
參考文獻--------------------------------------------------67
附錄------------------------------------------------------74
謝辭------------------------------------------------------75

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