順氯氨鉑引發的腎毒性在純系小鼠的確立與柴胡在此腎炎模型的藥效評估
中文摘要
臨床上，化學治療藥物順氯氨鉑 (cisplatin, CDDP) 常用於治療固體癌，其引起的腎毒性
，常是限制臨床使用的因素之一。本研究利用低劑量 CDDP 連續投予小鼠，引發腎炎 模型，以柴胡及 pentoxifylline (PTX) 作為預防藥物，評估其在此腎炎之預防效果。
實驗以 6 週齡雌鼠 (BALB/c mice, female) 投予不同劑量 CDDP (1.25, 2.5, 5 mg/kg/d) 及 分別投予 CDDP 1, 3, 5 天探討投予次數引起腎毒性的強度，以確立小鼠 CDDP 腎炎模 型。藉由事先投予柴胡濃縮劑 (BR 150, 300, 450 mg/kg/d) 及其純成分 saikosaponin A, C, D 、化學藥品 PTX, BR300 併用 PTX 來評估藥物對 CDDP 引起腎毒性的預防效果。實 驗以 N-acetyl-b-D-glucosaminidase (NAG) 、尿蛋白、 urinary creatinine (Ucr.) 與 BUN 評 估腎功能；其腎組織以 PAS 染色觀察病理組織及免疫螢光染色辨別其損傷部位之特異 性抗原。
實驗結果顯示，投予不同劑量與次數，以 CDDP 5 mg/kg/d 連續投予 5 天損傷最為嚴重
，尿液中 NAG 、尿蛋白與血清中 BUN 值增加， Ucr. 排除降低；組織學觀察到腎小管 有明顯的萎縮、脫落與圓柱體沈積，細胞浸潤較少。免疫螢光染色發現投予 CDDP 會 增加 TNF-a , cyclin D1 的表現。此外，投予 3 天 CDDP 即發現腎功能下降，組織學上 觀察到腎小管損傷。免疫螢光染色 TNF-a, cyclin D1 的表現，僅次於 5 天；但 p21 的表 現在投予 1 天時最強， 3 天次之， 5 天則明顯減弱。
投予藥物中，柴胡及其純成分 saikosaponin A, C, D 、化學藥品 PTX 及 BR300 併用 PTX 皆可改善腎功能，組織學上腎小管損傷有減輕的情形。藥效評估以 BR 300 mg/kg 預防 效果較好，純成分則以 saikosaponin D 最佳。
英文摘要
Cisplatin (CDDP) is an effective antineoplastic agent in the treatment of various solid tumors. The clinical utility of CDDP is limited because of its nephrotoxicity. We tried to establish CDDP induced nephrotoxicity in inbred mice and investigated the i mpact of Bupleuri Radix (BR) and pentoxifylline (PTX) on the model.
BALB/c mice (6 weeks, female) were intraperitoneally administered with 1.25, 2.5, 5 mg/kg/d of CDDP for 5 days (on day 1 t o day 5) to determine the experimental dose. Then, CDDP 5mg/kg/d were administered intraperitoneally to determine the relat ionship between the injection times (on day 1, 3, 5) and the degree of nephrotoxicity.
In the second experiment, CDDP (5 mg/kg/d) was given to mice on day 1 to day 5. The mice were administered orally with B upleuri Radix extract (BR) (150, 300, 450 mg/kg/d), saikosaponin A, C, D, PTX, BR300 combined with PTX once daily for 1 0 days (on day -5 to day 5). The control group was treated with distilled water.
Urinary N-acetyl-b-D-glucosaminidase (NAG), urinary protein, urinary creatinine (Ucr.) and blood urea nitrogen (BUN) were determined. Renal tissues were served to histological examination (PAS stain and immunofluorescence staining). The antibod ies including TNF-a, p21, cyclin D1 was chosen to recognize the specific antigens, which deposited in injury sites.
The results showed that animals administered with 5 mg/kg/d of CDDP for 5 days having the most severe injury. Elevation of NAG, urinary protein, BUN and decrease of Ucr were observed. The histological examination showed tubular atrophy, existe nce of cast and a little interstitial infiltration.
Immunofluorescent study revealed CDDP increased the deposition of TNF-a, cyclin D1 in kidneys. Besides, deterioration of k idney function and pathologic alterations were observed on day 3. The deposition of TNF-a, cyclin D1 on day 3 were increase d maximally to day 5 gradually. The expression of p21 increased maximally on day 1, but decreased on day 5.
The mice administered orally with BR300, saikosaponin A, C, D, PTX and BR300 combined with PTX ameliorated kidney d amage.
BR300 showed the strongest protective effect against CDDP induced nephrotoxicity among the BR groups. Saikosaponin D s howed the strongest protective effect among pure compound groups.
