| 摘要: | 台灣末期腎臟病(尿毒症)的發生率、盛行率皆高居世界前兩名，造成健保沉 重的負擔。馬兜鈴酸(aristolochic acid, AA)被認為是1993 年比利時馬兜鈴酸腎 病變(aristolochic acid nephropathy, AAN)事件中，造成減重婦女罹患AAN 及泌 尿道癌的主因，目前中西醫對AAN 尚無成熟的治療方法。臨床案例指出服用 大劑量馬兜鈴屬中草藥關木通會導致AAN，關木通於老鼠會致癌。本計畫擬以 關木通造成的小鼠AAN 為腎病動物模型，來探討健保前四大治療腎病的中醫 方劑(濟生腎氣丸、六味地黃丸、豬苓湯及知柏地黃丸)與單味藥(車前子、黃耆、 杜仲、丹參)對AAN 小鼠之治療效果與其作用機轉。 兩年計畫如下，第一年將研究健保四大中醫治腎病方劑對關木通致急性與慢 性小鼠AAN 的療效評估與其作用機轉。將藉助具高靈敏度活體即時性生物冷 光影像報導系統的NF-kB/FVB 基因轉殖鼠，加速監測連續暴露關木通7 天(或 12 週)所造成的急性(或慢性)AAN 發炎反應，並依冷光強度即發炎情形將老鼠 進行公平合理的分組，再以一倍臨床劑量的方劑連續治療AAN 小鼠7 天(或12 週)，以觀察中醫常用治療腎病方劑對AAN 小鼠之治療效果；觀察指標如下： 以周邊血彗星試驗法測量DNA 股斷裂情形，以微核測試法檢測遺傳毒性，以 血清生化指標(血尿素氮、血肌酐)、尿液分析(尿蛋白、尿肌酐)、腎組織切片(HE、 PAS、Masson 染色)、與免疫組織染色(中性?鏈內切?、轉化生長因子β1、結 締組織生長因子、纖維連結蛋白)來評估老鼠的腎毒性；部分12 週關木通暴露 FVB 野生型小鼠將飼養至瀕臨死亡，以觀察老鼠的存活率與腫瘤生成情形；每 一組小鼠(急性AAN 組、慢性AAN 組、腫瘤生成觀察組)犧牲後將進一步利用 基因晶片技術，分析AA 的靶器官腎臟與膀胱組織的基因表達圖譜變化情形， 以探討中醫方劑的療效作用機轉。第二年將探討健保前四大中醫治腎病單味 藥、與第一年實驗結果中具最佳療效的方劑之單味藥，對關木通致急性與慢性 AAN 小鼠的療效評估與其作用機轉，研究方法如同第一年。本計畫預計完成健 保前四大治療腎病的中醫方劑與單味藥對AAN 小鼠的療效評估與其可能作用 機轉，此結果可為臨床醫師應用此些中醫方劑與單味藥治療腎病提供實驗依 據，非常值得給中醫師作為臨床用藥參考。
Taiwan has the highest incidence and prevalence rates of Chronic kidney disease in the world. Aristolochic acid (AA), a known rodent carcinogen, is considered as a causative factor found in Aristolochia fangchi that causes urothelial carcinomas in patients with Aristolochic acid nephropathy (AAN). So far, there is no mature method to treat AAN in Western Medicine and Chinese Medicine. Caulis aristolochiae manshuriensis has been reported to cause AAN in patients and will induce AAN as well as carcinomas in rodents. In this study, we will use the noninvasive real-time NF-κB bioluminescence imaging technique and toxicogenomics to study the therapeutic mechanism of the top four most commonly used Chinese Medicine formulas for kidney disease in National Health Insurance Database and single drug on aristolochic acid nephropathy in mice. x In the first year of the study, the top four most commonly used Chinese Medicine formulas for kidney disease in National Health Insurance Database (Jisheng Shenqi Wan, Liuwei Dihuang Wan, Zu Ling Tang, Zhibai Dihuang Wan) will be studied for their therapeutic effects of on Caulis aristolochiae manshuriensis -induced nephrotoxicity and carcinogenicity in mice. The gene expression profile at target organs will be obtained by analyzing the function of these differentially expressed genes using Gene Ontology Consortium analysis and hierarchical method. The parameters of renal and liver function (the level of blood urea nitrogen, serum creatinine, urine creatinine, AST, ALT), the histopathology (HE, PAS, Masson staining) and immunohistochemistry staining (NF-kB and Neutral endopeptidase, transforming growth factorβ1, connective tissue growth factor, fibronectin) on renal and liver tissue sections in exposed mice will be examined. In the second year, the top four most commonly used Chinese Medicine single drug (Plantago asiatica L. etc) for kidney disease in National Health Insurance Database will be studied for their therapeutic effects of on Caulis aristolochiae manshuriensis-induced nephrotoxicity and carcinogenicity in mice. These data may provide a better understanding on relationship between Caulis aristolochiae manshuriensis-induced inflammation and cancer and the therapeutic effects of TCM on AA- and Caulis aristolochiae manshuriensis-induced AAN. These data will provide experimental evidences and guidelines for TCM doctors to treat kidney disease. |
