連翹乙醇萃取物之抗氧化及抗發炎活性

Antioxidant and anti-inflammatory activities of Forsythia suspensa ethanol extract

楊鎮鴻(Yang Cheng-Hung)

指導教授 ： 吳明昌

國立屏東科技大學/國際學院/食品科學國際碩士學位學程/碩士(2018年)

https://doi.org/10.6346/THE.NPUST.FS.001.2018.E11

摘要

連翹為亞洲常用的中草藥之一，為木犀科連翹屬植物的果實，富含許多化合物特別是連翹苷A，被證實具有抗菌，抗病毒，抗發炎及抗氧化等作用。因此本研究中，利用乙醇萃取物測試連翹抗氧化力並在人體真皮層細胞中測試萃取物之美白和抗發炎能力，經由初步分析後，連翹在65 %乙醇中浸泡24小時後並經減壓濃縮及冷凍乾燥萃取物有最佳抗氧化力，故本實驗以65 %乙醇為萃取濃度，在抗氧化結果顯示，連翹具有強力的清除DPPH及螯合亞鐵離子能力，且隨著濃度的上升抗氧化能力隨之上升，Nrf2為對氧化還原反應敏感之轉錄因子，此研究中發現連翹萃取物不但在體外具有強烈抗氧化作用，同時對細胞內也可促進Nrf2的活化，在美白及抗發炎的實驗則是以人類皮膚真皮層中的纖維母細胞 (CCD966SK) 並以脂多醣刺激，根據研究顯示纖維母細胞為人類皮膚中生成膠原蛋白的主要細胞，因此根據推斷促進纖維母細胞的生長，膠原蛋白也隨之增加進而促進皮膚的美白效果，在細胞存活率試驗中，結果顯示隨著連翹萃取物濃度增加，纖維母細胞也隨之增加，但在濃度250 (g/mL)下細胞有明顯減少，猜測因濃度太高使得連翹對細胞具有毒性反應，在後續實驗中，隨著連翹的濃度增加，具有抑制環氧化酶 (COX-2)蛋白表現作用，在其他促發炎因子介白素 (IL-6)、腫瘤壞死因子 (TNF-α) 及核因子活化B細胞κ輕鏈增強子 (NF-κB) 等皆具有抑制蛋白分泌作用，結果顯示，連翹在人體的細胞上具有抗發炎的作用，美白試驗的結果顯示，膠原蛋白的生成在脂多醣的刺激並無減少趨勢，然而在連翹萃取物的作用下有成長30% 的效果另外也具有抑制酪胺酸酶生成的作用，這將可作為往後美白及傷口感染治療產品開發參考。

關鍵字

連翹 ； 酪胺酸酶 ； 發炎 ； 抗氧化 ； 膠原蛋白 ； 纖維母細胞

並列摘要

Forsythia suspensa (Thunb.) Vahl, a commonly used supplement and alternative medicine, is a common Chinese medicine for gonorrhea, pyrexia and ulcer. It is reported to exhibit antipyretic and detoxification effects and has anti-bacterial, anti-virus and anti-inflammatory as well as high antioxidant activity. This study is to investigate the anti-inflammation of human fibroblasts of skin cells CCD966SK and the anti-oxidation capacity and inhibition tyrosinase effect by the extract of FS. The antioxidative activities were evaluated by various assays including DPPH radical, ferrous ions chelating test and TEAC assay. The results show 12, 19, 20, 33, 78 and 81% of scavenging DPPH radicals, 66, 79, 82, 116, 146 and 149% of chelating activity on ferrous iron with the increase of concentration enhance the ability. Nrf2 is a transcription factor sensitive to redox reaction. In this study, it has been found that FS extract not only has strong antioxidant activity in vitro but also up-regulated the expression of Nrf2 in CCD966SK. The fibroblasts of skin cell line CCD966SK were stimulated with LPS cocultured with ethanol extractions of FS. This experiment showed that the increase of the concentration of FS extract significantly reduced the formation of IL-6. FS extract was applied to of human skin cells CCD966SK to see whether it has a toxic effect on the cells in the cell survival test. It was found that with the concentration 250 g/mL the cell survival rate was decreased. Western blot analysis showed that FS extract significantly decreased the expression of inflammation factor and this effect was concentration dependent. The production of collagen in the stimulation of LPS did not reduce the trend. However, the effect of FS extract reduced growth effect and also has the role of inhibition of tyrosinase production.
The results showed that FS extract has significant antioxidative effect and ferrous iron chelating ability. The increase of concentration antioxidant capacity increase and inhibition tyrosinase ability also capacity has increased follow the concentration of extracts. The FS extract significantly decreased the expression of IL-6, NF-κB, TNF-α and COX-2 induced by LPS. In addition, FS extract can also increase the synthesis of collagen in fibroblasts.

並列關鍵字

Forsythia suspensa ； inflammation ； antioxidant ； collagen ； IL-6 ； fibroblast

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