| 摘要: | 「龍膽瀉肝湯」為中藥固有成方中最常被使用於治療肝臟疾病的方劑。本研究計畫旨在以大白鼠動物實驗配合微陣列晶片，初步探討長期服用「龍膽瀉肝湯」後對肝臟代謝酵素之影響。
經以每天三次連續灌食1gm/kg劑量之「龍膽瀉肝湯」達七天後，取出動物肝臟經一序列操作過程分離肝臟細胞微粒體，並萃取其中RNA，在經純化製成RNA樣品供生物晶片分析測定之用。晶片試驗採用ABC Rat UniversoChip 8K-1微陣列晶片，RNA試樣經螢光標誌(labeling)後與晶片上之cDNA雜交(hybridization)， 再以GenePix 4000B掃瞄[影像波長：635 nm (Treatment)及532 nm (Reference)]，最後以GenePix Pro 3.0.5.56專家軟體分析掃瞄結果。
由比對治療組與對照組在晶片上的基因表現，平均值比率大於2而具有基因表現調高(up-regulation)者共有約75種，其中較具生理意義的有：beta-globin、peptidylarginine deiminase type IV、hemoglobin alpha chain、hemoglobin alpha subunit、lysozyme、Phosphatidylinositol 4-kinase、phosphatidylinositol-4-phosphate 5-kinas、Thymosin beta-10、Topoisomerase II、TPA、Cathepsins、Chloride channel protein (p64)、farnesyl pyrophosphate synthase、ubiquitin-homology domain protein (PIC1)、cytoplasmic beta actin、UDP-glucose dehydrogenase及UDP-glucuronosyltransferase等。平均值比率小於0.5而具有基因表現調降(down-regulation)者共有約30種，其中較具生理意義的有：creatine kinase、Heparin-binding growth associated protein、NADH-ubiquinone oxidoreductase 及Syntaxin 2等。
有關長期服用「龍膽瀉肝湯」導致基因表現up-regulation 或down-regulation與生理活性之關連性有待繼續進一步探討，然而在本研究中並未發現對細胞色素P450各種代謝酵素有顯著的影響。
“Long-Dan-Shei-Gan-Tan”, a traditional Chinese herbal formulation, has been used for the treatment of hepatic-related diseases in the orient countries for more than thousand years. This study was aimed to primarily demonstrate the influence of long-term use of this formulation on the hepatic enzymes in rats by microarray biochip analysis.
The preparation was given orally trice a day with the dosage of 1 gm/kg for consecutive seven days. Livers were dissected and the microsomes were prepared by a series of separating processes. RNA samples were extracted and purified from microsomes, and then the fluorescence-labeled probes were hybridized with the immobilized cDNAs on the rat chips (ABC Rat UniversoChip 8K-1) and scanned with GenePix 4000B scanner ( and 532 were used for treatment and reference, respectively). The final images were eventually analyzed with the GenePix Pro 3.0.5.56 software.
The gene expressions of the “treatment” groups were compared with those of the “control” groups and an approximate of 80 genes were found up-regulated (Ratio of Means >2), among them, the genes with significant physiologic functions were beta-globin, peptidylarginine deiminase type IV, hemoglobin alpha chain, hemoglobin alpha subunit, lysozyme, Phosphatidylinositol 4-kinase, phosphatidylinositol-4-phosphate 5-kinase, Thymosin beta-10, Topoisomerase II, TPA, Cathepsins, Chloride channel protein (p64), farnesyl pyrophosphate synthase, ubiquitin-homology domain protein (PIC1), cytoplasmic beta actin, UDP-glucose dehydrogenase and UDP-glucuronosyltransferase; an approximate of 30 genes were found down-regulated (Ratio of Means <0.5), among them, the genes with significant physiologic functions were creatine kinase, Heparin-binding growth associated protein, NADH-ubiquinone oxidoreductase and Syntaxin 2.
The impacts on the pharmacological actions of long-term use of “Long-Dan-Shei-Gan-Tan” that leading to up-regulated or down-regulated gene expression should be further studied. However, the genes involved to cytochrome P450 enzymes were not found modified by the present formulation. |
