薄葉牛皮消抗氧化、抗菌與抗發炎活性之研究

Studies of Antioxidant, Antimicrobial and Anti-inflammatory activities of Cynanchum taiwanianum Yamazaki

李後易(Hou-I Lee)

指導教授 ： 楊季清 博士 ； 王子慶 博士

國立屏東科技大學/農學院/食品科學系所/博士(2012年)

https://doi.org/10.6346/NPUST.2012.00081

摘要

本研究目的係檢測薄葉牛皮消(Cynanchum taiwanianum Yamazaki) 抗氧化、抗菌性和抗發炎之效用。薄葉牛皮消屬蘿摩科 (Asclepiadaceae)，具有抗發炎、抗血栓及用於治療皮膚病、痢疾、便秘、肝炎、腎炎、白血球過多症、高血壓等功能。目前已知有五種主要成分化學立體結構包括乙醯苯 (acetophenones), 2,6-dihydroxyacetophenone (6), cynandione 1 (1), cynandione B-D (2-4), 及 cynanchone 5 (5)。由抗氧化能力分析結果薄葉牛皮消粉末以水、甲醇、乙醇60℃恆溫萃取15 min之萃取液，分析其DPPH (1, 1-dipheny-2-picryhydrazyl) 清除自由基能力、FRAP (Ferric reducing antioxidant potential assay)還原力、Total phenol 抗氧化能力表現最佳；乙醇萃取液在 25、40、60、80、100℃萃取均佳，甲醇萃取效果亦佳；水萃取液測 DPPH、FRAP 在 25、40、60、80、100℃均有穩定抗氧化能力效果。另以水、甲醇、乙醇各別在 60℃ 恆溫萃取 5、15、30、45 min 測 DPPH、FRAP、Total phenol 抗氧化能力表現；DPPH 在 25℃ 萃取液抗氧化能力亦佳，水萃取液 60℃ 15 min效果最好；還原力 (Reducting power) 結果，薄葉牛皮消粉末在低溫水或乙醇短時間加熱抗氧化能力為佳；總酚結果，以溫水服用宜佳，若以乙醇浸泡或加熱 (60℃ 15 min) 抗氧化能力均佳。抗菌活性篩選方面，以 Salmonella typhimurium 洋菜膠培養基 (Nutrient agar, NA)，使用 Streptomycin 0.1 g/mL、Ampicillin 0.1 g/mL 或薄葉牛皮消 0.1 g/mL測試，薄葉牛皮消粉末0.1 g/mL對 Salmonella typhimurium 有抑菌作用；再以Bacillus subtilis 洋菜膠培養基，使用Streptomycin 0.1 g/mL、Ampicillin 0.1 g/mL 或薄葉牛皮消 0.1 g/mL測試，發現薄葉牛皮消 0.1 g/mL 對 Bacillus subtilis 有抑菌效果。薄葉牛皮消粉末水和乙醇萃取物對鼠腎臟細胞株（NRK-52E）因IL-1β 所誘發之發炎作用之影響方面:水萃取物和乙醇萃取物不會影響NRK-52E 細胞生存力；同時水萃取物和乙醇萃取物對 NRK-52E 細胞在 IL-1β 誘發發炎之狀況下也不會影響 NRK-52E 細胞生存力。水萃取物和乙醇萃取物對 NRK-52E 細胞在 IL-1β 誘發發炎之狀況下可降低 NO 之生成量，結果顯示薄葉牛皮消粉末水和乙醇萃取物在以 IL-1β 誘發NRK-52E 細胞株發炎之狀況下，具有效抑制 NO、PGE2之生成，及原因可能與其顯著降低 p-IkB、NF-kB、iNOS、COX-2 蛋白質之表現有關。由三項實驗證明薄葉牛皮消具有抗氧化、抗菌性及對大鼠腎臟細胞抗發炎之效果。

關鍵字：薄葉牛皮消、抗菌、抗氧化、抗發炎、誘導型一氧化氮合成
酶、環氧化酶2、鼠腎臟細胞 (NRK-52E)。

關鍵字

薄葉牛皮消 ； 抗氧化、抗菌 ； 抗發炎 ； 誘導型一氧化氮合成 ； 環氧化酶2 ； 鼠腎臟細胞 (NRK-52E)

並列摘要

The major objective of this study was to antioxidation, antimicrobial activities and anti-inflammatory, Asclepiadaceae of Cynanchum taiwanianum Yamazaki (C.T). anti-inflammatory antithrombotic and mainly used as medicine for the treatment of skin disease, diarrhea, constipation, hepatitis, leukemia, hypertension herbs of effect, five components the stereochemistry include acetophenones, 2-6 dihydroxy-acetophenone (6), cynandione 1 (1), cynandione B-D (2-4), cynanchone 5 (5), has been known compound from C.T. The assay result of antioxidant effect shows following: to use the C.T powder extracts the essence by water, methanol and ethanol at 60℃ constant temperature for 15 min to examine the antioxidant ability of 1, 1-dipheny-2-picryhydrazyl (DPPH) Scavenging free radical effect, ferric reducing antioxidant potential assay (FRAP) reducting power and total phenol are the best; the ethanol extracts liquid well at 25, 40, 60, 80 and 100℃. Furthermore, it shows that has a study antioxidant effect result to examine DPPH , FRAP with the water essence at 25, 40, 60, 80 and 100℃, it means that the C.T should be taken with water. In addition, to use the water extracts, methanol and ethanol on 60℃ constant temperature in 5, 15, 30 and 45 min to examine the antioxidant effect of DPPH, FRAP and Total phenol; the DPPH extrats has a good antioxidant effect at the room temperature, and the effect upon water extrats at 60℃ for 15 min is the best; it has good antioxidant effect when the C.T powder heats in low-temperature water or the short time of ethanol; Total phenol should be good to take with the warm water. If soak or heat (at 60℃ for 15 min ) with the ethanol will has a good antioxidant effect. Screening of antimicrobial activity, to use 0.1 g/mL Streptomycin, Ampicillin and C.T powder to test with the Salmonella typhimurium in nutrient agar (NA), the result shows 0.1 g/mL C.T powder has antimicrobial function on Salmonella typhimurium; And then to use 0.1 g/mL Streptomycin, Ampicillin and C.T powder to test the Bacillus subtilis in NA, it shows the C.T has antimicrobial effect on Bacillus subtilis. The effect upon the Cynanchum taiwanianum water extracts (CTWE) and Cynanchum taiwanianum ethanol extracts (CTEE) to rat kidney cell (NRK-52E) cause the inflammation function from IL-1β. The water extracts and ethanol extracts will not affect the cell viability of NRK-52, and either to NRK-52E inducing under the inflammatory from IL-1β. Furthermore, the water extracts and ethanol extracts will reduce the products of NO under the inflammation function from IL-1β. These results indicate that Cynanchum taiwanianum ethanol extracts diminished IL-1 β -induced obviously reduces protein expression of reason possible and it p-IkB, NF-kB, iNOS and COX-2 (Cyclooxygenase 2) as well as subsequent production of NO and PGE2. It proved that has antioxidant, antimicrobial and anti-inflammatory of the C.T by three experiments.
Keywords: Cynanchum taiwanianum Yamazaki (C.T), Antioxidant,
Antimicrobial, Anti-inflammatory, iNOS, COX-2, NRK-52E cells.

並列關鍵字

Cynanchum taiwanianum Yamazaki (C.T) ； Antioxidant, Antimicrobial ； Anti-inflammatory ； iNOS ； COX-2 ； NRK-52E cells

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