含馬兜鈴酸類藥材其單味製劑與複方製劑於動物實驗之毒性研究

李水盛
台灣大學
中藥材的誤用，例如因為將減肥藥成份之一的防己科的粉防己(Stephania tetrandra)置換成馬兜鈴科的廣防己(Arsitolochia fangchi)而在歐洲導致在服用這類減肥藥的年輕婦人身上產生快速的腎臟間質細胞纖維化，稱為中藥性腎病變，是造成中藥中毒的主要原因之一。雖然Stephania tetrandra 與Arsitolochia fangchi皆可用來製備"防己"，但只有馬兜鈴科的 A. fangchi含有馬兜鈴酸，而馬兜鈴酸正是被認為造成此病變之主要成分。雖然已有許多關於馬兜鈴酸之研究，如腎毒性、致基因毒性和致癌性等，但卻鮮少人去研究經常被使用之馬兜鈴科中藥材或含馬兜鈴科藥材複方的毒性。因此在本計劃中我們將比較並探討馬兜鈴科植物馬兜鈴以及含馬兜鈴之複方藥材補肺阿膠湯之亞急性毒性並分析藥材及藥物處理動物內馬兜鈴酸及其代謝物之含量。在亞急性毒性測試中我們將觀察餵食這些藥材所造成之臨床表徵，血液、尿液生化值之變化與器官組織病理之改變，以作為肝、腎毒性之指標。而在化學成分及代謝物分析方面，為準確控制中藥材的給予量，本研究將以HPLC-UV定量實驗藥材中之aristolochic acid I及aristolochic acid II。至於其他含量較低的馬兜鈴酸衍生物，如aristolochic acid III、7-hydroxyaristilochic acid I、aristolochic acids IVa、aristolic acid等成分，則將以LC-MS進行定性分析。另外，本研究將取研究動物血清，定量其中之aristolochic acid I及aristolochic acid II及馬兜鈴酸代謝物aristolactam I，將個體吸收及代謝可能造成之變異考量在內。又因為馬兜鈴酸在動物模式及人體內的活化會受到一些酵素的表現量及活性的影響，在本計劃中將取研究動物肝及腎之cytosolic fraction，針對前述三種主要參與活化的酵素(CYP1A1、CYP1A2及NQD1)利用SDS-PAGE及Western blotting進行定量分析，觀察其表現量與腎毒性之相關性。透過化學及代謝分析，我們可以更進一步探討馬兜鈴酸及其代謝物與肝腎毒性之關聯性。 從以上實驗之結果，除了能進一步瞭解含馬兜鈴酸之藥材是否會造成與馬兜鈴酸一樣之毒性，我們更期盼提供一些安全性評估的資訊作為日後使用及管制中藥之參考。
關鍵字：中藥腎病；馬兜鈴酸

CCMP96-RD-044

Lee Shoei-Sheng
National Taiwan University
Misusage of medicinal herbs was considered one of the reasons that caused the intoxication in traditional Chinese medicine (TCM) administration. Inadvertently replacement of Stephania tetrandra by Arsitolochia fangchi, because both herbs are used as TCM under the same name of FangChi, has caused rapidly progressive interstitial renal fibrosis, named Chinese herbs nephropathy (CHN), in young women on a slimming regimen. Aristolochic acid, a constituent of A. fangchi but not S. tetrandra, was found in slimming regimen and was considered as the major component that caused the toxicity. Although much has been studied on the nephrotoxicity, mutagenecity and carcinogenesis of pure aristolochic acid, little information regarding to the commonly used medicinal herbs of Aristolochiaceae or the compound remedy containing Aristolochiaceae herb. Toxicity and chemical analysis of Aristolochiae Fructus (Ma-Do-Ling) and the compound remedy containing Aristolochiaceae herb Bu-Fe-Ya-Gia-Tong will be studied in this project. The subacute toxicity will be assessed by their ability to create changes in the clinical signs, blood and urine chemistry and gross anatomic examination after oral administration of the crude extract of the selected medicinal herbs. In order to accurately control the amount of herbs given to study animals.
關鍵字：Chinese herbs nephropathy；aristolochic acid