| 摘要: | 目前中藥影響細胞色素 P450 (Cytochrome p 450，CYP)的實驗大多聚焦在單一藥材萃取物或單一成分，然而中藥在使用時都是依照患者的體質或病症在方劑的基礎上進行藥材的疊加使用。因此了解藥材疊加對於CYP酵素的影響更貼近臨床使用。兩兩配伍的藥對是中藥組合的基礎，四君子湯為最常用的補氣方劑，因此研究目標為利用化學成分分析及對酵素活性與蛋白質表現量等方面討論四君子湯、異功散、六君子湯系列方劑與六種常用藥材之間的兩兩疊加對CYP3A4與CYP2C8兩種酵素的影響。在成分測定中，陳皮與甘草的總多酚與總黃酮含量較高。經過HPLC分析後，三種處方的化學成分組成為六種藥材的疊加，共煎後未出現明顯的新波峰。單味藥材中甘草、茯苓與陳皮，對於CYP3A4與CYP2C8時活性抑制較明顯 (CYP3A4 之 IC50分別為171.49 ?g/mL、526.71 ?g/mL、804.02 ?g/mL；CYP2C8 之 IC50分別為109.75 ?g/mL、1512.01 ?g/mL、1081.12 ?g/mL)；人參與白朮對於酵素活性影響不顯著；半夏於高劑量對CYP2C8活性有誘導潛力。方劑的部分，對CYP2C8的活性三種方劑的抑制率皆接近，對CYP3A4酵素活性，異功散與六君子湯些微高於四君子湯。CYP3A4酵素蛋白質表現量，人參、陳皮會明顯抑制CYP3A4表現，白朮則會誘導，茯苓、甘草與半夏的劑量依賴性較不明顯，四君子湯與六君子湯對CYP3A4酵素表現影響不明顯，異功散有較明顯的抑制表現。對小鼠同源酵素Cyp3a，只有白朮與甘草在較低劑量時會誘導表現，其餘藥材會抑制表現，四君子湯與異功散會抑制Cyp3a表現，六君子湯表現出了Cyp3a的誘導。五種藥材對CYP2C8蛋白質表現量影響皆不明顯，只有半夏在高濃度時有明顯的誘導作用，三種方劑都有些微的抑制效果，但劑量依賴性較不明顯。茯苓對Cyp2c有劑量依賴性的抑制表現，半夏無明顯影響，其餘四種藥材有誘導作用，方劑中只有四君子湯有誘導作用，其餘兩種為抑制表現。在藥對配伍部分探討了藥材兩兩配伍使用對CYP酵素活性和蛋白質表現量的影響，發現配伍後對活性抑制的效果主要是加成作用。表現量的結果顯示半夏在配伍其他藥材後對CYP3A4、CYP2C8與PXR表現量具有顯著的誘導作用。此外，陳皮作為CYP3A4的蛋白質表現抑制劑，其抑制效果在配伍其他藥材後會被緩解，並顯示出更高的酵素表現量。甘草在低劑量時可略微誘導CYP2C8表現，但配伍後則會顯著降低CYP2C8的表現量。中藥的使用涉及多種藥材的疊加，因此目前僅聚焦於單個藥材的中西藥交互作用研究是不夠完整的。未來應進一步驗證藥材疊加時對藥物動力學的影響，尤其是包含半夏、陳皮與甘草的組合，以提升用藥安全性。
Currently, most experiments on how traditional Chinese medicine (TCM) affects Cytochrome p 450 enzymes focus on single herbal extracts or single components. However, in clinical practice, TCM is used based on the patient's constitution or illness, with herbs being added to the basic formulas. Therefore, understanding the effects of herbal combinations on CYP enzymes is more clinically relevant. Two-herb combinations are the foundation of TCM formulas, with Si-Jun-Zi-Tang being the most commonly used formula for benefiting “Qi”. This study aims to utilize chemical composition analysis and enzyme activity and protein expression studies to discuss the effects of Si-Jun-Zi-Tang (SJZT), Yi-Gong-San (YGS), and Liu-Jun-Zi-Tang (LJZT) formulas and the pairwise combinations of six commonly used herbs on CYP3A4 and CYP2C8 enzymes.In the component analysis, tangerine peel (CP) and liquorice (GC) were found to have higher total polyphenol and total flavonoid content. HPLC analysis showed that the chemical composition of the three formulas was an overlay of the six herbs, with no significant new peaks observed after decoction.Regarding enzyme activity, GC, Poria (FL), and CP showed more significant inhibition on CYP3A4 and CYP2C8 (IC50 for CYP3A4 were 171.49 ?g/mL, 526.71 ?g/mL, and 804.02 ?g/mL, respectively; IC50 for CYP2C8 were 109.75 ?g/mL, 1512.01 ?g/mL, and 1081.12 ?g/mL, respectively). Panax Ginseng (RS) and Atractylodes (BZ) had no significant effect on enzyme activity, while Pinellia (BS) showed potential for inducing CYP2C8 activity at high doses. Among the formulas, the inhibition rates on CYP2C8 activity were similar, whereas for CYP3A4 activity, YGS and LJZT had slightly higher inhibition rates than SJZT.For CYP3A4 enzyme protein expression, RS and CP significantly inhibited CYP3A4 expression, while BZ induced it. The dose-dependency was not clear for FL, GC, and BS. SJZT and LJZT had no significant effect on CYP3A4 expression, whereas YGSn showed notable inhibition. For the mouse homolog enzyme Cyp3a, only BZ and GC induced expression at lower doses, while the other herbs inhibited it. SJZT and YGS inhibited Cyp3a expression, while LJZT induced it.The five herbs had no significant effect on CYP2C8 protein expression, except for BS, which showed notable induction at high concentrations. All three formulas had slight inhibitory effects on CYP2C8 expression, with no clear dose-dependency. FL showed dose-dependent inhibition of Cyp2c, while BS had no significant effect. The other four herbs induced Cyp2c expression, with only SJZT inducing expression among the formulas, while the other two inhibited it.The study on herb pairings explored the effects of pairwise combinations on CYP enzyme activity and protein expression, revealing that the inhibition effect on activity was mainly additive. The expression results showed that BS significantly induced the expression of CYP3A4, CYP2C8, and PXR when combined with other herbs. Additionally, the inhibitory effect of CP on CYP3A4 protein expression was alleviated when paired with other herbs, showing higher enzyme expression. GC slightly induced CYP2C8 expression at low doses but significantly reduced it when paired with other herbs.The use of TCM involves multiple herbs, making current research on single herb interactions with Western medicines incomplete. Future studies should further verify the effects of herbal combinations on pharmacokinetics, especially combinations including BS, CP, and GC, to enhance medication safety. |
