不同表現型牛樟芝的化學成分及免疫調節活性研究

Abstract

牛樟芝為一種高價值藥用真菌，其具有抗癌、抗氧化、降血糖、保肝、抗B型肝炎病毒、免疫調節及抗發炎等多種功效。自然界中除了常見的紅樟芝外，尚有甚少被研究的黃樟芝與白樟芝。本研究目的為確認不同表現型牛樟芝的基源，並分析其總三萜含量及組成分；另分析糙米培養之不同表現型牛樟芝的三萜類及多醣體含量，以及免疫調節活性。本研究共分析八種野外採集的不同表現型牛樟芝樣本。ITS區域定序結果確認這些不同表現型牛樟芝皆為牛樟芝物種，且已將其ITS序列登入NCBI的GenBank資料庫，其登入序號分別為MN947413、MN947415、 MN947416、MN947417、MN947418、MN947419、MK764936及MK764937。三萜類分析結果顯示，野外採集的紅樟芝、黃樟芝及白樟芝皆具有相同的三萜類組成分, 包括antcin A、antcin B、antcin C、antcin K、antcin G、antcin H 、dehydrosulfurenic acid、15-acetyldehydrosulfurenic acid及dehydroeburicoic acid，但紅樟芝的總三萜含量及其組成分的相對含量最高、黃樟芝次之、白樟芝最低。糙米培養之不同表現型牛樟芝中，紅樟芝與白樟芝的總三萜含量無明顯差異，且皆具有相似的三萜類組成分，唯有紅樟芝具有antcin C。多醣體物化性質分析結果表明，不同表現型牛樟芝多醣體皆具有相同的單醣組成，但兩者間葡萄糖、甘露糖、阿拉伯糖及岩藻糖含量卻有差異；此外，紅樟芝多醣體的分子量也較白樟芝小。雖然紅樟芝及白樟芝多醣體皆會抑制經LPS誘導之RAW264.7巨噬細胞的TNF-α及IL-6生成量，然而僅白樟芝多醣體能顯著抑制NO生成量。綜上所述，紅樟芝具有最高的總三萜含量；白樟芝多醣體的抗發炎活性較紅樟芝佳，說明多醣體物化性質差異會影響其抗發炎活性強弱。

Antrodia cinnamomea (AC) is a high value medicinal mushroom with various bioactivities such as anticancer, antioxidative, hypoglycemia, hepatoprotection, anti-hepatitis B virus, immunomodulation and anti-inflammation. In nature, besides the commonly known red AC phenotypes, there are also yellow and white AC phenotypes that are not well studied. The aims of this study were to authenticate the phylogenetic identity of different AC phenotypes, and analyze their triterpenoid contents and compositions, followed by investigating the triterpenoids and polysaccharides contents and immunomodulatory activities of different brown rice cultured AC phenotypes. Eight different wildly collected AC phenotypes were subjected to phylogenetic analysis to confirm their authenticity. Based on ITS sequence results, the eight different AC phenotypes were confirmed to be species AC, and their DNA sequence have been registered in NCBI GenBank with the following accession numbers: MN947413, MN947415, MN947416, MN947417, MN947418, MN947419, MK764936 and MK764937. Among the different AC phenotypes, results showed that red AC had the highest triterpenoid contents, followed by yellow AC and white AC. HPLC profiles indicated that different AC phenotypes had similar triterpenoid compounds, including antcin A, antcin B, antcin C, antcin K, antcin G, antcin H, dehydrosulfurenic acid, 15-acetyldehydrosulfurenic acid and dehydroeburicoic acid. For different AC phenotypes cultured on brown rice, results showed that red AC and white AC had similar triterpenoid contents, but their triterpenoid profiles were slightly different. Only antcin C was found in red AC. The polysaccharides of both red AC and white AC cultured on brown rice possessed similar monosaccharide compositions; however, the glucose, mannose, fucose, and arabinose contents between them are different. In addition, the molecular weight of red AC polysaccharides was much lower than white AC. The polysaccharides from red AC and white AC significantly inhibited the TNF-α and IL-6 production in LPS-induced RAW264.7 macrophage cells; however, only white AC polysaccharides was found to significantly inhibit NO production. In conclusion, the results indicate that among different AC phenotypes, red AC had the highest triterpenoid contents, and the differences in the physicochemical properties of white AC polysaccharides may lead to its better anti-inflammatory potency than red AC.