| 摘要: | 最近以中草藥或健康食品來治療或預防疾病的觀念則越來越受到重視。探討防風通聖散(Bofutsushosan; BOF)及大柴胡湯(Daisaikoto; DAI)兩種傳統中草藥方劑對國人保健應有其重要的意義。民間多認為防風通聖散(BOF)及大柴胡湯(DAI)具有保肝、減肥及降血脂等效能。但是防風通聖散(BOF)及大柴胡湯(DAI)兩種方劑之免疫調節的活性與其保健功效目前並不清楚，因此值得深入探討。本研究主要研究目的以LPS-誘導小鼠(Raw 267.4)細胞探討BOF及DAI之免疫調節活性及其分子作用機制，含TLR-4、MyD88、MAPK及NF-κB等途徑之訊息傳遞反應；並以動物實驗模式探討BOF及DAI進行免疫調節反應包含細胞吞噬活性、細胞激素及抗體量測定分析之影響。另外，也透過細菌逆轉突變性分析(Ames test)及30天毒性試驗對BOF及DAI進行安全性評估。研究結果發現脂多醣(LPS)誘導後，以25~50 μg/mL的防風通聖散及大柴胡湯的乙醇萃取物對Raw264.7細胞能有效的促進細胞增生(P<0.05)。在管外及管內實驗結果發現以LPS誘導後，防風通聖散及大柴胡湯的乙醇萃取物均會明顯降低IL-1β、IL-6及IL-10之產生量。另外，大柴胡湯的乙醇萃取物對小鼠巨噬細胞會降低IFN-γ之表現量。細胞以LPS誘導後加入防風通聖散及大柴胡湯乙醇萃取物均會降低TLR4、MyD88及TRAF6 mRNA之表現量(p<0.05)及抑制細胞核NF-κB之表現，但對MAPK訊息分子之傳遞路徑不影響。在30天毒性試驗，結果發現高劑量防風通聖散及大柴胡湯乙醇萃取物均會減少小鼠腎相對體重及降低小鼠增加體重，而血清生化測定結果發現餵養25 mg/kg大柴胡湯的乙醇萃取物可顯著降低血糖，總膽固醇及BUN濃度。另外， Ames試驗結果發現防風通聖散及大柴胡湯乙醇萃取物對沙門氏菌(TA100)均無致突變性。由Ames試驗及生化檢測結果發現防風通聖散及大柴胡湯乙醇萃取物均無毒性。防風通聖散及大柴胡湯乙醇萃取物會增加淋巴細胞之增生作用及對小鼠巨噬細胞能顯著的增加CD40之表現。
There is growing interest in the use of traditional herbal medicines and health foods for the treatment and prevention of diseases. Herbal medicines, such as Bofutsushosan (BOF) and Daisaikoto (DAI) possess hepatoprotective, anti-obesity, and hypolipidemic activities among other therapeutic properties. However, their immuno-regulatory effects remain unclear, which warrant research investigation. In the second year, we examine the immunomodulatory activities of BOF, DAI, and their respective active compounds to clarify their effects on molecular signaling cascades, such as TLR-4, MyD88, MAPKs, and NF-κB pathways in LPS-induced mouse macrophage cells (Raw 264.7). In addition, we examine the effects of BOF and DAI on the immune responses (i.e. phagocytosis, cytokine production) in animal systems. We also evaluate the safety of BOF and DAI through bacterial reverse mutation assay (Ames test) as well as the 30-day toxicity test in mice. Results showed that BOF and DAI ethanol extracts significantly prevented LPS-induced cell cytotoxicity in murine macrophage (Raw264.7) cells. 10 to 50 μg/mL BOF and DAI ethanol extracts effectively increased the cell proliferation in a dose-dependent pattern. BOF and DAI ethanol extracts significantly down-regulated IL-1β, IL-6 and IL-10 levels in vitro and in vivo. In addition, our data showed that DAI ethanol extract treated-group effectively inhibited LPS-induced IFN-γ production in murine macrophages. 50 μg/mL BOF and DAI ethanol extracts significantly blocked TLR4, MyD88 and TRAF6 mRNA levels (P<0.05), and nuclear factor-κB (NF-κB) expression, but did not affect the MAPK pathway. 50 mg/kg BOF and DAI ethanol extracts treated-mice had decreased kidney relative weight and decreased body weight gain in the 30-day toxicity test. Results showed that 25 mg/kg DAI ethanol extract treated-group had significantly lower serum glucose, total cholesterol, and BUN levels compared to the control group in the biochemical assay. In the Ames test, BOF and DAI ethanol extracts displayed no mutagenicity toward the strain (Salmonella typhimurium TA100) tested and no remarked toxic effects were found in the Ames test and biochemical assay. Furthermore, BOF and DAI ethanol extracts enhanced lymphocyte proliferation and increased the expression of CD40 in murine macrophages. |
