利用馬兜鈴酸腎症腎炎模型評估綠茶及(+)-catechin的藥效

陳世銘
台北醫學大學
近年來發生許多馬兜鈴酸腎病變 (aristolochic acid nephropathy, AAN)的案例，其原因是誤用含馬兜鈴酸的植物。腎臟受損部位侷限於近曲小管，有明顯的細胞浸潤、腎小管萎縮，以及間質纖維化的情形。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，並評估綠茶(green tea, GT)、(+)-catechin (CAT)對AAN的改善效果。
純系小鼠C3H/He (6weeks, males) 連續56天飲水給予3.0 μg/mL AA，之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56天AA後，連續14天經口投予GT 75, 150, 300 mg/kg，CAT 25, 50, 100 mg/kg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，並進行免疫螢光染色，包括：巨噬細胞，乙型轉型生長因子(TGF-β)，基質金屬蛋白分解酶(MMP-9)，甲型腫瘤壞死因子(TNF-α)，以辨識損傷部位之特異性抗原。
關鍵字：馬兜鈴酸腎病變，綠茶，(+)-catechin，巨噬細胞，乙型轉型生長因子，基質金屬蛋白分解酶，甲型腫瘤壞死因子

Effect of Green Tea and (+)-catechin on aristolochic acid-induced nephropathy in inbred mice

Shih-Ming Chen
Taipei Medical University
Many cases about aristolochic acid nephropathy (AAN) have been identified in recent years. The reason is that people mistaken the herb that contains aristolochic acid (AA). AAN is a unique type of nephropathy, which was characterized by extensive cell infiltration, tubular atrophy, and interstitial fibrosis. We established a chronic interstitial fibrosis model of AAN in inbred mice and investigated the efficacy of green tea (GT), (+)-catechin (CAT) on AAN.
AA will be dissolved in distilled water (3μg/ml) and as drinking water ad libitum to C3H/He mice (6 weeks, male) for 56 days, the control group will be administered to distilled water. In the second experiment, they will be administered orally with GT (75, 150, 300 mg/kg), CAT (25, 50, 100 mg/kg) once daily for 14 days. The control group will be administered to distilled water.
Urinary protein (UP), urinary N-acetyl-beta-D-glucosaminidase (NAG), blood urea nitrogen (BUN), blood glucose will determine. Renal tissues will serve 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), TNF-α (tumor necrosis factor-α) will be chosen to recognize the specific antigens, which deposited in injury sites.
All animals will be treated with AA developed increased in UP, NAG, BUN, and blood glucose. In the histological examination, we will observe tubular atrophy, interstitial infiltration and fibrosis in the typical AAN. In the immunofluorescence stain assay, macrophage, TGF-β, MMP-9, and TNF-α will localize in the renal tissue.
關鍵字：Aristolochic acid nephropathy (AAN), green tea, (+)-catechin, macrophage, TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9), TNF-α (tumor necrosis factor-α)