Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/51810
標題: 靈芝與柳松菇之抗氧化性質和其對腫瘤細胞之毒性及柳松菇之抗致突變性質
Antioxidant Properties and Their Cytotoxic Activities on Tumor Cells of Ganoderma tsugae and Agrocybe cylindracea and Antimutagenic Properties of A. cylindracea
作者: 蔡淑瑤
Tsai, Shu-Yao
關鍵字: 靈芝;Ganoderma tsugae;柳松菇;抗氧化性質;抗致突變性;癌細胞毒性;Agrocybe cylindracea;antioxidant properties;antimutagenic properties;anti-tumor activity
出版社: 食品科學系
摘要: 
本研究係針對成熟靈芝(Ganoderma tsugae Murrill,松杉靈芝)(六星期採收)及靈芝茸子實體(二星期採收)、菌絲體和濾液與柳松菇(Agrocybe cylindracea Murrill)子實體、菌絲體和濾液進行一般成分分析,再將甲醇及熱水萃取所得萃取物,進行抗氧化性質和天然的抗氧化成分之測定及評估其對腫瘤細胞之影響。同時對柳松菇子實體、菌絲體和濾液熱水萃取物評估其抗致突變性。
靈芝、靈芝茸、菌絲體和濾液甲醇萃取物在抗氧化性質方面,靈芝和靈芝茸甲醇萃取物有較佳抗氧化力。還原力部分,靈芝、靈芝茸、菌絲體和濾液甲醇萃取物在 5 mg/ml時,分別為 0.50、0.93、1.05和 0.95。靈芝和靈芝茸甲醇萃取物有較佳的清除DPPH自由基活性,於 5 mg/ml時,分別為 88.39 和 93.81%。螫合亞鐵離子能力方面,於 10 mg/ml時,依序為靈芝茸(91.16%)~ 濾液(93.31%)>菌絲體(85.87%)>靈芝(66.21%)。
靈芝和靈芝茸熱水萃取物之抗氧化力較佳,在 20 mg/ml 時,分別為 78.50 和78.20%。靈芝、靈芝茸、菌絲體和濾液熱水萃取物,在還原力部分於 5 mg/ml時,分別為 1.08、1.04、0.95 和 1.12。在清除 DPPH 自由基方面,菌絲體之清除效果最佳(20 mg/ml,91.23%)。在清除羥自由基方面,於 20 mg/ml時,依能力大小為靈芝(72.27%)~ 靈芝茸(73.69%)>菌絲體(55.88%)>濾液(45.96%)。
柳松菇子實體甲醇萃取物具有極佳抗氧化力(5 mg/ml,90.03%)。在還原力及清除DPPH自由基能力部分,依序為子實體>濾液>菌絲體。螫合亞鐵離子能力部分,在 5 mg/ml時,子實體、菌絲體及濾液分別為 90.56、84.58 及 96.34%。
柳松菇子實體、菌絲體及濾液熱水萃取物在抗氧化力部分,於 20 mg/ml時,依序菌絲體(81.63%)>子實體(63.55%)>濾液(56.78%)。在還原力部分,在 20 mg/ml時,依序為濾液(1.14)>子實體(1.02)>菌絲體(0.86)。在清除DPPH自由基方面,在 10 mg/ml時,依序為濾液(73.53%)>菌絲體(68.19%)>子實體(65.89%)。
在抗氧化成分分析上,總多酚類化合物是所有試驗樣品中含量最多的天然抗氧化成分。
在抗致突變部分,實驗使用 Ames test 方法,以 Salmonella typhimurium TA97、TA98、TA100 及 TA102試驗菌株,使用直接致突劑 NQNO(不加S9混合物)與間接致突劑 B[a]P(加S9混合物)來探討柳松菇子實體、菌絲體及濾液熱水萃取物抗致突變性。結果顯示,所使用之樣品在 0.05、0.1、0.5、1及5 mg/plate劑量範圍下,不具毒性和致突變性,且柳松菇子實體、菌絲體及濾液熱水萃取物皆具抗致突變性,其中以柳松菇子實體熱水萃取物在 TA97 對 B[a]P 抗突變效果最佳。
在腫瘤細胞毒性試驗,以 MTT 染色法觀察 C6 、 Hep 3B 和 HL-60 癌細胞生長情形。靈芝、靈芝茸、菌絲體和濾液甲醇萃取物部分,以靈芝及靈芝茸對 HL-60 癌細胞株有較佳抑制癌細胞生長能力(在 0.2 mg/ml,癌細胞存活率分別為 31.86及16.50%)。而在熱水萃取物部分,對 C6 、 Hep 3B 和HL-60癌細胞株生長之抑制能力,隨著濃度增加而增加,且抑制能力大小依序為靈芝>靈芝茸>濾液>菌絲體。
柳松菇子實體、菌絲體及濾液甲醇萃取物對 C6、 Hep 3B 和HL-60癌細胞生長情形,隨著劑量增高存活率有些微下降,其中三者之間對癌細胞株存活率沒有差異。熱水萃取物部分,對 C6、Hep 3B 和 HL-60癌細胞生長情形,隨著劑量增高存活率有下降趨勢。

This research used mature and baby Ling chih, mycelia and fermentation filtrate of Ganoderma tsugae Murrill and fruit bodies, mycelia and filtrate from the submerged culture of Agrocybe cylindracea Murrill to study the proximate composition, antioxidant properties and anti-tumor activity of methanolic and hot water extracts. The antimutagenic activity of hot water extracts from A. cylindracea in the form of fruit bodies, mycelia and filtrate from submerged culture.
Antioxidant properties of G. tsugae were studied in the form of methanolic extracts from mature and baby Ling chih, mycelia and fermentation filtrate; the methanolic extracts from mature and baby Ling chih showed high antioxidant activities. The reducing powers of methanolic extracts were 0.50, 0.93, 1.05 and 0.95 at 5 mg/ml for Ling chih, baby Ling chih, mycelia and filtrate, respectively. The methanolic extracts from Ling chih and baby Ling chih scavenged 1,1-diphenyl-2-picrylhydrazyl radicals by 88.39 and 93.81% at 5 mg/ml, respectively. At 10 mg/ml, chelating effect were in the order of filtrate (93.31%) ~ baby fruit bodies (91.16%) > mycelia (85.87%) > mature fruit bodies (66.21%).
The hot water extracts from Ling chih and baby Ling chih antioxidant activities by 78.50 and 78.20% at 20 mg/ml, respectively. The reducing powers of hot water extracts were 1.08, 1.04, 0.95 and 1.12 at 5 mg/ml for Ling chih, baby Ling chih, mycelia and filtrate, respectively. The hot water extracts from mycelia showed good ability of scavenged 1,1-diphenyl-2-picrylhydrazyl radicals. At 20 mg/ml, Scavenging effect on hydroxyl radicals were in the order of Ling chih (72.27%) ~ Baby Ling chih (73.69%) > mycelia (55.88%) > filtrate (45.96%).
The methanolic extracts from A. cylindracea fruit bodies showed an excellent antioxidant activity (90.03%) at 5 mg/ml. At 5 mg/ml, reducing powers were in the order of fruit bodies (0.99) > filtrate (0.50) > mycelia (0.35). At 5 mg/ml, scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radicals were in the similar order of fruit bodies (93.78%) > filtrate (74.19%) > mycelia (61.80%). At 5 mg/ml, all methanolic extracts showed excellent chelating effects and were 90.56, 84.58 and 96.34% for fruit bodies, mycelia and filtrate, respectively.
The antioxidant activity of hot water extracts were in the order A. cylindracea mycelia (81.63%) > fruit bodies (63.55%) > filtrate (56.78%). At 20 mg/ml, reducing powers were in the order of filtrate (1.14) > fruit bodies (1.02) > mycelia (0.86). The hot water extracts from A. cylindracea at 10 mg/ml, scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radicals were in the order of filtrate (73.53%) > mycelia (68.19%) > fruit bodies (65.89%).
Total phenols were the major naturally occurring antioxidant components found in all samples.
The hot water extracts from A. cylindracea fruit bodies, mycelia and filtrate have been tested for their antimutagenic properties against direct-acting mutagen of NQNO (without S9 mix) and indirect-acting mutagen of B[a]P (add S9 mix), using the Salmonella typhimurium tester strains TA97, TA98, TA100 and TA102. Five different dosage were used, which were 0.05, 0.1, 0.5, 1.0, 5.0 mg/plate. All the tested extracts had no toxicity and mutagenicity. The hot water extracts from A. cylindracea fruit bodies, mycelia and filtrate showed best antimutagenicity effect. The hot water extracts from A. cylindracea fruit bodies showed excellent effect against B[a]P toward TA97.
In the anti-tumor activity test, the inhibitory effects on C6, Hep 3B and HL-60 tumor cell lines were studied using MTT test. The methanolic extracts from Ling chih and baby Ling chih had the highest inhibitory activity for HL-60 tumor cell line (31.86 and 16.50% inhibition at 0.2 mg/ml).The inhibitory effects on C6, Hep 3B and HL-60 tumor cell lines of hot water extracts increased with the increasing concentration of hot extracts. The order of inhibitory activity of hot water extracts is mature Ling chih > Baby Ling chih > filtrate > mycelia.
The inhibitory effects on C6, Hep 3B and HL-60 tumor cell lines of methanolic extracts from A. cylindracea fruit bodies, mycelia and filtrate slowing increased with the increasing concentration of hot water extracts. The order of inhibitory activity of methanolic extracts is similar. The inhibitory effects on C6, Hep 3B and C6 tumor cell lines of hot water extracts from A. cylindracea fruit bodies, mycelia and filtrate increased with the increasing concentration of hot extracts.
URI: http://hdl.handle.net/11455/51810
Appears in Collections:食品暨應用生物科技學系

Show full item record
 

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.