小葉桑根抗氧化及抗發炎活性之研究

Study of anti-oxidative and anti-inflammatory bioactivity of root of Morus australis

張哲愷(Che-Kai Chang)

指導教授 ： 曾翠華

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2014年)

https://doi.org/10.6834/CSMU.2014.00024

摘要

小葉桑 (Morus australis Poir)是桑科植物，過去被使用於治肺熱咳嗽、關節炎、風濕病的治療，有文獻指說其莖與根含有酪胺酸酶抑制劑，具有美白的活性，但抗氧化及抗發炎等功效及其作用機制，尚未闡明。我們初步對小葉桑的根與莖微波萃取並比較其組成，發現小葉桑根微波萃取物(MA)具有較多的皂苷類與多酚類成分，推測可能具有保健預防的特性。首先我們對MA進行HPLC分析，證實含有morusin以及oxyresveratrol等多酚類成分，除此之外還有萜類如Oleanolic acid，並且做了試管實驗發現其具有清除DPPH及NO的能力。接著利用脂多糖 (LPS)誘發老鼠巨噬細胞的模式(RAW-264.7 cell)，發現MA可抑制LPS誘導的nitrite、PGE2、IL-6、TNF-α、ROS等發炎指標，在蛋白質分析上也看到相同的結果，且MA這些作用是抑制STAT1與MAPK路徑有關。進一步動物實驗，以先餵食MA處理再使用CCl4 誘使小鼠發炎，經7週由組織切片上發現低濃度的MA可以促進肝組織的細胞分裂，而高濃度不僅可以促進細胞分裂，更能減少CCl4所造成的細胞發炎與壞死。另一方面也發現MA也可抑制CCl4所誘發小鼠肝組織的脂質過氧化與IL-6的分泌，進而從蛋白質分析上也可看到可抑制CCl4誘導的iNOS、COX-2表現，並使HO-1表現增加。綜合以上可確定MA具有抗氧化、抗發炎的作用進而有保肝的效果。針對MA裡的主成分morusin、oxyresveratrol、oleanolic acid進行細胞實驗分析，發現皆可抑制LPS所誘導RAW 264.7細胞nitrite的分泌，且對morusin深入研究發現其也能抑制LPS誘導的PGE2、IL-6、TNF-α的釋放與ROS的累積。在蛋白質分析上，也擁有抑制LPS誘導的iNOS、COX-2表現，並促進HO-1表現，因此morusin也具有抗氧化、抗發炎的效果。
以上結果顯示，MA具有抗氧化、抗發炎的作用，且其作用認為與其富含多酚類與三萜類有關。

關鍵字

脂多糖 ； 前列腺素E2 ； HO-1

並列摘要

Morus australis Poiris a mulberry plant,which has been used to treat hyperactivity cough, arthritis, rheumatism in the past. The literature reports Morus australis stems and roots containing tyrosinase inhibitors, which showes whitening effect. But its anti-oxidation and anti-inflammatory effects has not been well elucidated. First, we study of Morus australis root and stem by microwave extraction and compare its composition. It showed microwave extract of Morus australis root (MA) contained more saponin and phenolic components, which may exhibit health prevention properties. In the present study, we used a macrophages model of mouse (RAW-264.7 cell) to observe whether MA possesses antioxidant and anti-inflammatory potential.Second, we analyzed by HPLC for MA,confirmed that it does havepolyphenols: morusin and oxyresveratrol and other ingredients such as oleanolic acid, a triterpenoid. In the preliminary study, it showed MA have ability to scavenge DPPH and NO. In cell experiments, MA inhibited LPS-induced nitrite, PGE2, IL-6, TNF-α and ROS production in RAW-264.7 cells. In the protein analysis, it showed MA inhibited LPS-induced expression of iNOS, COX-2, but promoted expression of HO-1.The signal pathway is associated with STAT-1 and MAPK pathway.In animal experiments, we fed MA and used CCl4 inducing inflammation in mice and found MA not only reduced CCl4 inducing cellular inflammation but also promoted liver cell division in tissue sections. MA also inhibited lipid peroxidation and accumulation of IL-6 induced by CCl4 in mouse liver induce. By protein analysis of liver tissue, MA inhibited iNOS, COX-2 and promotes HO-1 expression in analysis of proteins.Based on the above it implicates MA exhibits hepatic protection effect by its anti-oxidant and anti-inflammatory property. In addition, the main ingredients of the MA such as morusin, oxyresveratrol, oleanolic acid showed inhibitory effect in the LPS-induced NO production in RAW264.7 cells. Further research found morusin also inhibited LPS-induced production of PGE2, IL-6, TNF-α and ROS in RAW264.7 cells. In the protein analysis,morusin also suppressed LPS-induced iNOS, COX-2, but promoted expression of HO-1.
In conclusion, MA possesses antioxidant and anti-inflammatory effect, which is related with MA containing polyphenols and triterpenoids.

並列關鍵字

Lipopolysaccharide ； PGE2 ； HO-1

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