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Title: 草魚和蛇膽汁中膽酸鹽和膽醇鹽組成及其在大白鼠中之保健和毒作用探討
Studies on compositions of bile salt and bile alcohol in the grass carp and snake bile juice, and their healthy function and toxicity in the rats
Authors: Yen-Hung Yeh
葉彥宏
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: ;毒性;草魚;
bile;toxicity;grass carp;snake
Date: 2003
Issue Date: 2011-06-30T07:39:50Z
Abstract: 我國民間有人相信吞食草魚膽、鯉魚膽、蛇膽和雞膽等動物膽汁具有明目、清熱解毒和止咳平喘等功效,但在臨床上卻發現民眾因生食草魚膽汁和鯉魚膽汁中毒而導致急性腎衰竭現象。國內亦曾發生長期攝食蛇膽而導致腎衰竭之案例。由於動物膽汁之成分,如膽醇鹽和膽酸鹽等皆有毒性,而且因動物不同其膽汁組成分亦不同,因此所引起之腎衰竭及其他症狀,如胃腸和神經方面症狀亦有所不同。其中草魚膽汁之真正致毒成分;以前國內外一直懷疑是膽醇和膽酸鹽所致,本研究室已證實係由膽醇鹽5□-cyprinol sulfate所致;並已開發了高效能液相層析法 (HPLC) 同時分析十數種魚類膽醇鹽和膽酸鹽之成分,亦證明不同魚種,其膽汁中之膽醇鹽和膽酸鹽組成不同,也發現蛇膽和草魚膽之膽酸鹽組成和毒作用截然不同。由於陸上動物膽汁廣被國人食用或製成中藥劑,而且蛇膽干亦曾引起使人致死事件,因此本研究利用已開發之HPLC方法,首先分析草魚及不同蛇膽汁之膽酸鹽成分,得知草魚膽汁主要以5□-cyprinol sulfate為主,雨傘節 (Bungarus multicinctus) 膽汁主要以cholic acid和chenodeoxycholic acid為主;眼鏡蛇 (Naja naja atra) 膽汁主要以cholic acid和chenodeoxycholic acid為主;龜殼花 (Trimeresurus mucrosquamutus) 膽汁主要以cholic acid和taurodeoxycholate為主;赤尾青竹絲 (Trimeresurus stejnegeri) 膽汁主要以cholic acid、chenodeoxycholic acid和taurodeoxycholate為主。其次,分析其他陸上動物後,發現豬、牛和鵝膽汁主要以cholic acid 和chenodeoxycholic acid 為主,羊膽汁主要以cholic acid和 deoxycholic acid為主。另外,並分析由中藥房採購安宮牛黃丸、六神丸、牛黃清心丸和三鞭至寶丸等中藥劑後,發現其中並無含有膽酸鹽之成分。 另外,為知草魚膽汁中的毒性物質對於大白鼠的短期毒性,乃以24隻Wistar系大白鼠,分成4組,每隔3天分別以口胃管餵予生理食鹽水、草魚膽汁 (40 mg/0.6 ml)、5□-cyprinol (40 mg/0.6 ml) 和5□-cyprinol sulfate (40 mg/0.6 ml) 飼養期間為19天。在短期毒性試驗中,草魚膽汁所產生的症狀和5□-cyprinol sulfate相似。為了解pentoxifylline (PTX) 對食用草魚膽汁所引起大白鼠中毒之治療情形,以18隻Wistar系大白鼠,分成3組,每隔3天分別以口胃管餵予生理食鹽水、5□-cyprinol sulfate (40 mg/0.6 ml) 和5□-cyprinol sulfate (40 mg/0.6 ml),飼養期間為19天,第19天以PTX (12 mg/0.6 ml saline) 腹腔注射連續4天。結果發現PTX對於草魚膽汁引起之腎衰竭有治療之作用,對於肝臟損傷則沒有治療作用。 利用以HPLC分析所得草魚和眼鏡蛇之膽汁酸組成份,以標準品調配成草魚膽汁之組成和草魚膽汁之主要成份5□-cyprinol sulfate,對大白鼠進行短期毒性試驗。即以24隻Wistar系大白鼠,分成4組,每隔3天分別以口胃管餵食生理食鹽水、5α-cyprinol sulfate/ chenodeoxycholic acid (9:1) (40 mg/0.6 ml)、5α-cyprinol sulfate (40 mg/0.6 ml) 和chenodeoxycholic acid (40 mg/0.6 ml),飼養期間為19天。在短期毒性試驗中,以標準品調配的草魚膽汁所產生之症狀和5□-cyprinol sulfate相似。另以標準品調配成眼鏡蛇膽汁酸組成份,對大白鼠進行短期毒性試驗。即以24隻Wistar系大白鼠,分成4組,每隔3天分別以口胃管餵予生理食鹽水、一顆蛇膽之組成份 [cholic acid (19.5 mg), chenodeoxycholic acid (20 mg), deoxycholic acid (2.7 mg), lithocholic acid (4.8 mg), taurochenodeoxycholate (8.7 mg) 和 glycolithocholate (7.2 mg)/3 ml] × 1、二顆蛇膽之組成份及三顆蛇膽之組成份,飼養期間為19天,結果發現,隨著劑量的增加,會產生蓄積性的毒作用,引起肝、腎功能的病變。 為了解草魚膽汁對肝功能是否有補強的影響,將48隻Wistar系大白鼠分成兩大組,分別是對照組和肝傷害組,各大組再分4小組,每隔3天分別以口胃管餵予cholic acid (10 mg/0.6 ml)、cholic acid (20 mg/0.6 ml) 和5α-cyprinol sulfate (10 mg/0.6 ml),飼養期間為29天。在短期毒性試驗中,以acetaminophen腹腔注射 (150 mg/kg, i.p) 連續4天。結果發現在第5和17天其肝功能指標AST、ALT 和 ALP 有明顯之上升的情況,在第29天時對照組和肝傷害組之肝功能指標之間並無差異性,顯示草魚膽補肝並無顯著之功效。 為了解草魚膽汁是否有補強明目之功能的影響,將48隻Long-evans系大白鼠分成兩大組,分別是對照組和糖尿病組,各大組再分4小組,每隔3天分別以口胃管餵予cholic acid (10 mg/0.6 ml)、cholic acid (20 mg/0.6 ml) 和5α-cyprinol sulfate (10 mg/0.6 ml),飼養期間為29天。在短期毒性試驗中,以streptozocin腹腔注射 (60 mg/kg, i.p)。在飼養第29天時分析其血液和眼球內維生素A的含量,發現對照組和糖尿病組的眼睛功能指標之間確有差異性,但草魚膽並不會增進維生素A之生合成,顯示草魚膽對於明目並無顯著之功效。
Some Chinese people believe that eating the animal gallbladder, such as bile of grass carp (Ctenopharyngodon idellus), snake and chicken, may improve their visual acuity. However, poisoning cases have occasionally occurred when people ingested animal bile, especially grass carp bile. Among those cases, the major symptom of victims was acute renal failure. Disturbance of liver, gastrointestinal tract and central nervous system were also noted. The major toxic material of grass carp bile juice is 5α-cyprinol sulfate. On the other hand, the toxicity and component of snake and grass carp bile juice are different. In the mean time, the determination method for bile acids in fish bile juices was developed. It was found that high performance liquid chromatography (HPLC), with a reversed ODS column and a buffer system of 0.3% ammonium carbonate/acetonitrite (73:27, v/v) 10 min→(68:32) 10 min→(50:50) 10 min, clearly separated 14 bile acids. Based on the analyses of above HPLC method, it was found that the major component of the bile juices of the bile juice from different snake were as follows:cholic acid and chenodeoxycholic acid in Bungarus multicinctus and Naja naja atra;cholic acid and taurodeoxycholate in Trimeresurus mucrosquamutus;cholic acid、chenodeoxycholic acid and taurodeoxycholate in Trimeresurus stejnegeri. The porcine, cattle and goose bile juice was mainly cholic acid and chenodeoxycholic acid, the sheep bile juice was mainly cholic acid and deoxycholic acid, other 4 Chinese medicines were not found to contain any species bile acids. To compare the short-term toxicity of toxic components of grass carp bile juice (GCBJ) in rats. Twenty-four male Wistar rats were divided into 4 groups and treated orally every 3-days with 40 mg each of freeze-dried GCBJ powder, 5α-cyprinol and 5α-cyprinol sulfate for 19 days. The grass carp bile juice exhibited significant toxicity, and the short-term toxicity of 5α-cyprinol sulfate and GCBJ powder was similar to each other. To investigate the effect of pentoxifylline (PTX) on the toxicity of grass carp bile juice in rats, 18 male Wistar rats were divided into 3 groups and treated orally every 3-days with 40 mg each of 5α-cyprinol sulfate and 5α-cyprinol sulfate for 19 days. PTX also counteracted the renal failure induced by grass carp bile. However, the liver injury caused by grass carp bile was not prevented by PTX. To elucidate the toxic effect of grass carp bile acids, a synthetic bile acid mixture was prepared mimicking the bile acid composition of a grass carp bile juice and used in rats. Attempts are made to elucidate the effect of bile acid chenodeoxycholic acid on the toxicity of bile alcohol 5α-cyprinol in rats. Twenty-four male Wistar rats were divided into 4 groups and treated orally at 3-days periodic treatment with each 40 mg each of 5α-cyprinol sulfate/chenodeoxycholic acid (9:1), 5α-cyprinol sulfate and chenodeoxycholic acid for 19 days. After treated with 5α-cyprinol sulfate/chenodeoxycholic acid (9:1), 5α-cyprinol sulfate and chenodeoxycholic acid, the toxicity of 5α-cyprinol sulfate/chenodeoxycholic acid (9:1) and 5α-cyprinol sulfate was similar to each other, and the toxic effect of chenodeoxycholic acid was less. In bile juice of snake gallbladder, cholic acid and chenodeoxycholic acid were predominant components in 3 snake species. To elucidate the toxic effect of snake bile acids, a synthetic bile acid mixture was prepared mimicking the bile acid composition of a snake Naja naja atra bile juice. Twenty-four male wistar rats were divided into 4 groups and treated orally at 3 days periodic treatment with saline solution (control group) and different doses (1-3 folds) of the bile acid mixture. It indicated that short-term toxicity of snake Naja naja atra bile acids was significant in rats. To investigate the remedial effect of grass carp bile acid on liver function, male wistar rats were used as experimental animals. The liver damaged rats induced by acetaminophen (150 mg/kg, i.p) for 4 days. The experiment was divided into two groups (with or without acetaminophen). Each group (24 rats) were divided into four subgroups (6 rats). The rats in each subgroup were treated orally at 3-days periodic treatment with saline solution (control group), 10 mg of cholic acid, 20 mg of cholic acid, and 10 mg of 5α-cyprinol sulfate, respectively. Fresh diets and drink water were available and ad libitum for 29 days. The activities of AST, ALT, and ALP in the plasma of rats treated with acetaminophen group in the 5 and 17 days were significantly increased. But after 29 days, the activities of AST, ALT and ALP in the rats if acetaminophen group were different from those of control group. The bile acid and bile alcohol did not seem to ameliorate the damage of liver function induced by acetaminophen. To investigate the remedial effect of grass carp bile acid on vision function, male Long-evans rats were used as experimental animals. The diabetic rats induced by streptozocin (60 mg/kg, i.p) and experiment was divided into two group (with and without diabetic). Each group (24 rats) was then divided into four subgroups (6 rats). The rats in each subgroups were treated orally at 3-days periodic treatment with saline solution (control group), 10 mg of cholic acid, 20 mg of cholic acid, 10 mg of 5α-cyprinol sulfate, respectively. Fresh diets and drink water were available and ad libitum for 29 days. The levels of vitamin A in the plasma and eyes of rats treated without and with diabetic treatment after 29 days were significantly different. It meant that bile acid and bile alcohol did not seem to ameliorate the vision function of eyes.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G00000JENOB
http://ntour.ntou.edu.tw/ir/handle/987654321/14562
Appears in Collections:[食品科學系] 博碩士論文

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