馬兜鈴酸腎症在純系小鼠的確立與柴胡在此腎炎模型的藥效評估

許光陽
中國藥學會
馬兜鈴酸腎病變 (aristolochic acid nephropathy, AAN)。其受損部位侷限於近曲小管，除了明顯的間質纖維化，腎小管萎縮，還有輕微細胞浸潤的情形。本研究藉由長期投予純系小鼠低劑量aristolochic acid (AA) 成功誘發與人類AAN相似的腎炎，並評估柴胡 (Bupleuri Radix) 及saikosaponin A, C, D等對AAN的改善效果。純系小鼠C3H/He (6weeks, males) 連續28天飲水給予AAs後停藥，恢復正常飲用水觀察14天，對照組在實驗期間投予正常飲用水。藥效評估實驗依前述方法給予28天AAs後，連續14天經口投予柴胡濃縮劑 (BR) 150, 300, 450 mg/kg，柴胡成分saikosaponin A, C, D，延胡索、葛根湯，對照組則給予蒸餾水。測定尿蛋白，N-acetyl-beta-D-glucosaminidase (NAG) 與血中尿素氮 (blood urea nitrogen, BUN)，來評估小鼠腎功能；腎組織使用PAS染色觀察病理組織改變，及進行免疫螢光染色 (巨噬細胞，TGF-β，MMPs) 以辨識損傷部位之特異性抗原進行分析。 利用上述的方法，期待能夠確立馬兜鈴酸腎病變模型，同時可評估柴胡藥效在此腎炎模型。關鍵字: 馬兜鈴酸腎病變，柴胡，巨噬細胞，乙型轉型生長因子，基質金屬蛋白分解酶
關鍵字：馬兜鈴酸腎病變；柴胡；巨噬細胞；乙型轉型生長因子；基質金屬蛋白分解酶

Establishment of the aristolochic acid nephropathy in inbred mice and effect of Bupleuri Radix on the nephritis

Kuang-Yang Hsu
The Pharmaceutical Society of China Taiwan
Aristolochic acid nephropathy (AAN) is a unique type of nephropathy, which was characterized by an extensive interstitial fibrosis with rare cell infiltration and tubular atrophy. We tried to establish a chronic interstitial fibrosis model of AAN in inbred mice and investigated the impact of Bupleuri Radix, saikosaponin A, C, D on AAN. Aristolochic acid (AA) dissolved in distilled water were administered orally to C3H/He mice (6 weeks, male) as drinking water ad libitum for 4 weeks, control group were treated with distilled water. In the second experiment, they were administered orally with Bupleuri Radix extract (BR) (150, 300, 450 mg/kg), saikosaponin A, C, D , Corydalis Yanhusuo Rhizoma and Gee Gen Tong ,once a day for 14 days. The control group was treated with distilled water. Urinary protein, urinary N-acetyl-beta-D-glucosaminidase (NAG), blood urea nitrogen (BUN), blood glucose were determined. Renal tissues were served to histological examination (PAS stain and immunofluorescence staining). The antibodies, including murine F4/80 macrophage, TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9) was chosen to recognize the specific antigens, which deposited in injury sites. Aristolochic acid nephropathy was established in inbred mice, and the toxicity effect of Bupleuri Radix on the nephritis. Key word: Aristolochic acid nephropathy (AAN), Bupleuri Radix, macrophage, TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9)
關鍵字：Aristolochic acid nephropathy (AAN)；Bupleuri Radix；macrophage；TGF-β(transforming growth factor-β)；MMP-9 (matrix metalloproteinase-9)