臺灣博碩士論文加值系統

在漢方中，桑黃是一種珍貴的藥用真菌，指的是寄生於桑樹等植物上的多年生硬菇類的總稱。桑黃在國外研究中具有多種保健效果。桑黃之研究主要於韓國及日本，在台灣研究極少。原產地有日本、韓國等東南亞地區、澳洲、北美及中國等。
第一部份實驗以三種桑黃品系Phellinus igniarius (火木層孔菌)、Phellinus linteus (裂蹄木層孔菌)、Phellinus pini (松木層孔菌) 為材料，對其進行一般組成分、礦物質含量、機能性成分分析與安全性試驗。桑黃水萃取物安全性試驗以Ames test方法進行，Salmonella typhimurium TA98及TA100為試驗菌株，以直接致突變劑NQNO(-S9) 與間接致突變劑AFB1(+S9) 進行其抗致突變性試驗。結果顯示，一般組成分中，P. igniarius與P. linteus 水分含量15.95、15.2 % 較高，粗纖維含量P. pini 含量11.76 % 最高。礦物質含量P. linteus 鈣含量 11.81 % 最高，P. linteus與P. pini 鉀含量13.06 %、14.66 % 最高。機能性成份方面，P. igniarius、P. linteus與P. pini多醣的含量分別是4.98 %、2.54 %和3.82 %，P. igniariussβ-1,3-glucan的含量有4.34 mg/g dry weight，P. linteu為5.29 mg/g dry weight，P. pini含量為6.16 mg/g dry weight，SOD-like活性分別P. igniarius 119.05 U/ml、P. linteus 69.23 U/ml和P. pini 59.00 U/ml。三種品系桑黃於安全性試驗 Ames test 測
定下，在0.5~10 mg/plate 沒有毒性和致突變性。
第二部份實驗以桑黃品系P. pini (松木層孔菌) 其水萃取物利用動物模式針對四氯化碳 (CCl4) 誘導大鼠肝損傷，針對化學性肝損傷進行護肝功能之評估。主要分為正常對照組、負對照組 (CCl4 肝損傷組)、正對照組 (Silymarin治療對照組)、高劑量組100 mg/kg、中劑量組50 mg/kg、低劑量組 10 mg/kg。結果顯示，在血液中超氧歧化酶 (superoxide dismutase, SOD)、GOT (glutamate oxaloacetate transaminase)、GPT (glutamate pyruvate transaminase) 值皆有下降趨勢，並且在麩胱甘肽過氧化酶 (glutathione peroxidase, GPx) 及麩胱甘肽還原酶 (glutathione reductase, GRd) 的數據可看出其正對照組及實驗組高、中、低劑量皆可提升其含量，且明顯與負對照組有顯著性差異。從整體結果可知，P. pini萃取物中劑量組 50 mg/kg的護肝效果較佳，因此在護肝方面可以搭配其它具有護肝效果之素材，開發為保健食品。

In traditional Chinese herbal medicine, Sang-Hwang is a kind of valuable medicinal mushroom. It is a group of perennial hard mushrooms which are parasite on mulberry and so forth. At overseas researches, it has been found many kinds of health care effects. Most of Sang-Hwang researches have been studied in Korea and Japan, and extremely few researches in Taiwan. Its origin is distributed from Japan, Korea to China, Australia, and North America.
In chapter one, three kinds of Sang-Hwang strains, Phellinus igniarius、Phellinus linteus and Phellinus pini were used as material to analyze proximate composition, content of microelements and content of functional components. In antimutation test, Salmonella typhimurium TA98 and TA100 are experiment bacterial strains for testing Sang-Hwang water extracts by Ames test with direct mutation agent NQNO(-S9) and indirect mutation agent AFB1(+S9). The results showed that content of functional components, P. igniarius, P. linteus and P. pini polysaccharide content was 4.98 %, 2.54 % and 3.82 %, respectively. β- 1,3- glucan content of P. linteus was 5.29 mg/dw g, P. igniarius 4.34 mg/dw g, and P. pini 6.16 mg/dw g. In SOD-like activity, P. igniarius was 119.05 U/ml, P. linteus 69.23 U/ml and P. pini 59.00 U/ml. On the other hand, there were no toxicity and mutation reaction at 0.5~10 mg/plate for three kinds of Sang-Hwang.
In chapter two, efficiency evaluation of P. pini protecting liver was in animal mode aimed at rat chemistry liver damage inducing with carbon tetrachloride. It was divided into normal group, carbon tetrachloride group (CCl4 liver damage group), Silymarin treatment group, high dosage group 100 mg/kg, middle dosage group 50 mg/kg, and low dosage group 10 mg/kg. The results showed that superoxide dismutase (SOD), glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminas (GPT) all were downside. Through contents results of glutathione peroxidase (GPx) and glutathione reductase (GRd), the Silymarin treatment group and the experiment groups high, middle, low dosage had promoted effect, and significantly differenced from carbon tetrachloride group, obviously. From these whole knowable results, P. pini water extracts protecting liver effect were better in dosage 50 mg/kg than others. For this result, P. pini can combine with other protecting liver materials to develop as health food.

目次

摘要............................................................................................................i
Abstract.....................................................................................................iii
目次............................................................................................................v
圖次............................................................................................................x
表次...........................................................................................................xi


第一章、文獻回顧………………………………………………………1
一、桑黃簡介……………………………………………………………3
（一）Phellinus igniarius (L.:Fr.) Quél. 火木層孔菌......................3
1.生物活性…………………………………………...………4
(1) 動脈舒張……………………………………………….4
(2) 抗腫瘤功效…………………………………………….4
(3) 抗致突變功效………………………………………….4
（二）Phellinus linteus (Berk. et Curt.) Teng 裂蹄木層孔菌.........6
1.生物活性…………………………………………………...7
(1) 調節免疫功能……………………………….....…........7
(2) 抑制腫瘤生長及轉移………………………………….7
(3) 抗血管新生、抗發炎活性………………………….....8
(4) 護肝作用……………………......…………….…...........8
(5) 治療關節炎………………………......………………....8
(6) 誘導細胞凋亡………………......………………............8
（三）Phellinus pini (Brot.:Fr.) Ames 松木層孔菌……………….11
二、機能性成份…………………………………….…………………..14
β 1-3 Glucan………………………………………….…………..14
三、Ames Test……......……………….......……......……………….......17
（一）原理.………….......……......…......………….………..........17
（二）藥物轉化酵素 S9….......……......…......………………….18
四、水飛薊素 Silymarin….......……......…......………………………..18
五、四氯化碳 CCl4……......…......……………………………………..19

第二章、桑黃之一般組成、機能性成份含量分析及安全性評估…..22
摘要……………………………………………………………………..23
前言……………………………………………………………………..24
實驗架構………………………………………………………………..25
一、材料與方法………………………………………………………..26
（一）樣品……………………………………………………………..26
（二）樣品製備………………………………………………………..26
（三）試藥……………………………………………………………..26
（四）儀器…………………………………………………….………..28
二、一般組成分分析…………………………………………….……..30
（一）水分含量測定…………………………………….………..30
（二）粗蛋白含量測定………………………………….………..30
（三）粗脂肪含量測定……………………………….…………..31
（四）粗纖維含量測定……………………………….…………..31
（五）粗灰分含量測定………………………………….………..32
（六）無氮萃取物含量之測定……………………………………32
三、礦物質含量分析………………………………………….………..32
四、機能性成份分析…………………………………………….….….32
（一）總酚含量測定…………………………………….………..33
（二）類黃酮含量測定…………………………………….……..33
（三）粗多醣含量測定…………………………………………...34
（四）總醣含量測定……………………………………………...34
（五）β-1,3-Glucan 含量測定…………………….…………….35
（六）類超氧歧化酶活性分析……………………………………36
五、安全性試驗Ames test………………………………………………36
（一）毒性試驗……………………………………………………36
（二）致突變性試驗………………………………………………38
（三）抗致突變性試驗……………………………………………40
七、統計分析……………………………………………………………42
結果討論………………………………………………………………..43
結論……………………………………………………………………..47


第三章、餵食桑黃(Phellinus pini)萃取物對四氯化碳誘導肝損傷之護肝效果…………………………………………………………………..62

摘要……………………………………………………………………..63
前言……………………………………………………………………..64
實驗架構………………………………………………………………..65
一、材料與方法…………………………………………….…………..66
（一）樣品……………………………………………….………..66
（二）試藥……………………………………………….………..66
（三）儀器……………………………………………….………..67
二、試驗動物…………………………………………………….……..68
三、飼育管理……………………………………………….…………..68
四、護肝試驗……………………………………………………………69
（一）四氯化碳誘發肝損傷………………………………………69
（二）血液及肝臟樣品前處理……………………………………70
（三）血清轉胺酵素活性分析……………………………….…. 61
（四）抗氧化酵素活性分析……………………………………...61
（五）肝臟脂質過氧化分析………………………………………72
（六）麩胱甘肽相關酵素活性分析………………………………72
（七）蛋白質定量分析……………………………………………72
（八）肝組織切片染色觀察………………………………………72
五、統計分析……………………………………………………………73
結果討論………………………………………………………………..74
結論……………………………………………………………………..78
總結論…………………………………………………………………..88
參考文獻………………………………………………………………..89
















圖次
圖1-1、火木層孔菌....................................................................................5
圖1-2、桑黃對癌細胞作用機制...............................................................9
圖1-3、裂蹄木層孔菌.............................................................................10
圖1-4、松木層孔菌.................................................................................12
圖1-5、具有抗腫瘤活性的 β-(1,6) 分支的β-(1,3)-D-葡聚醣結構…15
圖1-6、投予β-glucan後產生的寄主免疫反應.....................................16
圖1-7、四氯化碳肝毒性的機制.............................................................20
圖1-8、四氯化碳代謝途徑.....................................................................21
圖3-1、動物於實驗期間之體重趨勢圖................................................81
圖3-2、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷之肝臟組織病理的影響...................................................................................87



















表次
表1、利用帶有Sarcoma 180腫瘤細胞的老鼠進行熱水萃取液的抗腫瘤性篩選.........................................................................................13
表2-1、桑黃之ㄧ般組成成份...............................................................48
表2-2、桑黃之礦物質含量...................................................................49
表2-3、桑黃水萃取之萃取率...............................................................50
表2-4、桑黃之總酚類及類黃酮化合物含量.......................................51
表2-5、桑黃之機能性成分含量...........................................................52
表2-6、Phellinus igniarius 萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之毒性試驗(±S9) ...................................................................53
表2-7、Phellinus linteus 萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之毒性試驗(±S9) .......................................................................54
表2-8、Phellinus pini 萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之毒性試驗(±S9)…………………………………………..……..55
表2-9、Phellinus igniarius萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之致突變性試驗(±S9)……………….……………………...56
表2-10、Phellinus linteus萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之致突變性試驗(±S9)…………………………………...…….57
表2-11、Phellinus pini萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之致突變性試驗(±S9)……………………………………..…..…58
表2-12、Phellinus igniarius 水萃取物對於NQNO(-S9)及AFB1(+S9)對鼠傷寒沙門桿菌TA98及TA100菌株之抗致突變性試驗……………………………..….……………………………..…59
表2-13、Phellinus linteus 水萃取物對於NQNO(-S9)及AFB1(+S9)對鼠傷寒沙門桿菌TA98及TA100菌株之抗致突變性試驗…………………………………………………..……………...60
表2-14、Phellinus pini 水萃取物對於NQNO(-S9)及AFB1(+S9)對鼠傷寒沙門桿菌TA98及TA100菌株之抗致突變性試驗……………………………………..………….…………….….61
表3-1、桑黃水萃取物對四氯化碳誘導急性肝損傷之大鼠體重及肝臟重量變化之影響.............................................................................80
表3-2、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷之血清轉胺酵素含量.............................................................................................82
表3-3、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷之SOD及Catalase活性影響……………………………………….…….….83
表3-4、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷之血清膽固醇含量…………………………………..………………….………..84
表3-5、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷紅血球之GPx及GRd活性影響……………………………………...………….85
表3-6、桑黃水萃取物對四氯化碳誘發大鼠急性肝損傷肝臟之GPx及GRd活性影響………………………………………….…………86

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