| 摘要: | 小燈籠草（Kalanchoe gracilis Hance）為景天科（Crassulaceae）燈籠草屬植物的全草。民間用於解熱、鎮痛、抗發炎等，為常用民間藥，其葉形如雞爪又名雞爪?帘峖W大返魂。
本研究以小燈籠草莖部的甲醇萃取物及經有機溶媒分劃得之正己烷層、氯仿層、乙酸乙酯層、正丁醇層及水層等萃取物。體外抗氧化進行，DPPH自由基清除能力試定(1,1-diphenyl-2-picrylhydrazyl)，還原力測定、Trolox等價抗氧化能力（Trolox equivalent antioxidant capacities, TEAC）分析、總多酚及類黃酮含量測定等試驗。細胞抗發炎，以LPS誘導RAW 264.7生成一氧化氮（nitric oxide, NO）與誘導型一氧化氮合成?﹛]Inducible nitric oxide synthases, iNOS）及cyclooxygenase (COX-2)表現等方式評估抗氧化活性。以肝癌G2細胞評估抑制增生之活性。
抗氧化活性評估結果顯示小燈籠草莖部之TEAC抗氧化能力以乙酸乙酯層為最佳，總還原力及DPPH自由基清除能力則以氯仿層為最佳。總黃酮及總多酚含量以乙酸乙酯層及氯仿層高於其他分層萃取物。細胞抗發炎方面，小燈籠草莖部氯仿層萃取物可明顯抑制LPS誘導RAW 264.7生成一氧化氮（nitric oxide, NO）與誘導型一氧化氮合成?﹛]Inducible nitric oxide synthases, iNOS）及cyclooxygenase (COX-2)表現。抑制Hep G2細胞增生試驗，以氯仿層萃取物抑制細胞增生作用最佳。抗氧化試驗採用之標準品eupafolin也有很好的抗氧化及抑制細胞增生能力。綜合上述結果可能與小燈籠草莖部總多酚含量有關，黃酮類eupafolin可能是重要的活性成分之一。
動物實驗部分，以醋酸扭體試驗及福馬林舔足試驗探討口服小燈籠草莖部甲醇萃取物之鎮痛作用，以λ-角叉菜膠誘發足蹠浮腫，探討口服小燈籠草莖部萃取物之抗發炎作用，並測定小鼠肝組織之超氧歧化?﹛]superoxide dismutase, SOD）、麩胱甘?汐棜??﹛]glutathione reductase, GRd）及麩胱甘?此L氧化?﹛]glutathione peroxidase, GPx）活性之變化，探討其抗發炎作用機轉。鎮痛實驗結果，小燈籠草莖部甲醇萃取物對於醋酸誘導扭體反應，有明顯減少扭體次數之作用，以及福馬林誘發舔足反應之後期有抑制小鼠的舔足時間。抗發炎實驗結果，小燈籠草莖部甲醇萃取物（0.5 g/kg與1.0 g/kg），對於λ-角叉菜膠誘發足蹠浮腫亦有明顯的抑制效果。λ-角叉菜膠誘導三小時後，小燈籠草莖部甲醇萃取物增加肝臟組織SOD及GRd活性及減少腫脹腳掌(malondialdehyde, MDA)濃度。小燈籠草莖部甲醇萃取物會減少λ-角叉菜膠誘導增加的Interleukin-1 beta (IL1-β), Tumor necrosis factor alpha (TNF-α)及NO濃度。
結果顯示小燈籠草莖部甲醇萃取物具有鎮痛及抗發炎作用。小燈籠草莖部甲醇萃取物抗發炎作用機轉可能與經由增加肝組織SOD及GRd活性及減少腫脹腳掌IL1-β，TNF-α及NO濃度而減少腫脹腳掌MDA濃度有關。
Kalanchoe gracilis Hance is a medicinal plant from the Crassulaceae family. It is called folklorically “Da-Huan-Hun” in Chinese. It is also referred to as “Ji Zhao Huang”, because the morphology of its leave is look-alike chicken claws. In Taiwan, it is a folk medicine commonly used by traditional Chinese medicine (TCM) practitioners for treating pain, fever and inflammation.
The aim of this study was to evaluate the antioxidant, anti-inflammatory and antiproliferative activities of the methanolic extract and fractions from the stem of K. gracilis by using DPPH radical scavenging test, reducing power, trolox equivalent antioxidant capacity (TEAC), total phenolic compounds and total flavonoid contents, LPS-induced nitric oxide (NO) production in RAW264.7 cell, and Hep G2 cell proliferation.
Among all fractions, Ethyl-Acetate fraction showed the highest TEAC activities, Chloroform fraction showed the highest Reducing power and DPPH radical scavenging activities. The Chloroform fraction also had highest contents of polyphenol and flavonoid. Chloroform fraction also decreased the LPS-induced NO production and expressions of iNOS and COX-2 in RAW264.7 cells. Chloroform fraction had the highest antiproliferative activity Eupafolin also had good pharmacology activity in the antioxidant, anti-inflammation and antiproliferation. Eupafolin might be an important bioactive compound in the stem of K. gracilis. The above experimental data indicated such efficacy may be mainly attributed to its polyphenolic compounds.
In this study, we also evaluated the analgesic effect of the methanol extract of K. gracilis stem (MKGS) in animal models by inducing writhing response with acetic acid and formalin test. The anti-inflammatory effect of MKGS was also estimated on mice with λ-carrageenan induced paw edema model. In order to investigate the anti-inflammatory mechanism of MKGS, we analyzed the activities of glutathione peroxidase (GPx) and glutathione reductase (GRd) in the liver, and the levels of interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α) , malondialdehyde (MDA) and nitric oxide (NO) in the edema paw. In the analgesic tests, MKGS (0.5 and 1.0 g/kg) decreased both the acetic acid-induced writhing response and the licking time in the late phase of the formalin test. In the anti-inflammatory test, MKGS (0.1, 0.5 and 1.0 g/kg) decreased paw edema at the third, fourth, fifth and sixth hours after λ-carrageenan had been administrated. Furthermore, MKGS increased the activities of SOD and GRd in liver tissues and decreased MDA level in the edema paws three hours afterλ-carrageenan was injected. MKGS also affected the levels of IL-1β, TNF-α and NO induced by λ-carrageenan.
All these results suggested that MKGS possessed analgesic and anti infla- mmatory effects. The anti-inflammatory mechanism of MKGS might be related to the lowering of MDA level in the edema paw via increasing the activities of SOD and GRd in the liver, as well as the decreases in the levels of TNF-α and NO, and IL-1β, and the production of MDA in inflamed tissues. |
