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  • 學位論文

D型甲硫胺酸對順鉑誘發大鼠毒性作用之影響

Effects of D-methionine on cisplatin-induced toxicity in rats.

指導教授 : 歐珠琴
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摘要


順鉑(cis-diamminedichloroplatinum(II), Cisplatin)是臨床上廣泛使用的化療藥物,但cisplatin嚴重的副作用是導致使用受限的原因之一,這些嚴重的副作用包含腎毒性、腸胃毒性、神經毒性、耳毒性等器官損傷。D型甲硫胺酸(D-methionine)為含硫的必需氨基酸,具有抗氧化能力,且不影響cisplatin的抗腫瘤活性。D-methionine降低cisplatin造成耳毒性的相關研究已廣泛被探討,但對腎毒性及腸胃毒性的研究相當的少。因此本研究想要探討D-methionine是否可降低cisplatin所誘導的毒性傷害。首先以CCK-8偵測D-methionine與cisplatin共同培養對人類腎臟近端腎小管細胞(HK-2細胞)的毒性作用,以西方墨點法及RT-PCR分析cleaved caspase 7之蛋白表現量及AQP1之mRNA表現量,結果顯示,D-methionine可增加HK-2細胞之存活率、AQP1之mRNA表現量及降低cleaved caspase 7之蛋白表現量。動物實驗部分,將大鼠隨機分配成三組(每組7隻):(1)控制組,每週腹腔注射0.9% NaCl並每天灌食一般飲用水(2)cisplatin組,每週腹腔注射5 mg/kg cisplatin並每天灌食一般飲用水(3)cisplatin + D-methionine組,每週腹腔注射5 mg/kg cisplatin並每天灌食300 mg/kg D-methionine。於施打三劑cisplatin後犧牲。實驗結果發現,cisplatin + D-methionine組與cisplatin組相比,顯著增加平均體重、攝食量、平均紅血球容積(MCV)、血小板數量及血鉀濃度(p<0.05);顯著降低平均紅血球血色素濃度(MCHC)腎臟重量/體重比值、腎臟脂質過氧化產物-丙二醛(MDA)的生成(p<0.05),但無法減輕血清肌酐(serum creatinine)及尿素氮(BUN)之上升。本研究顯示,D-methionine可減緩cisplatin所誘導的體重減輕及改善厭食,因此D-methionine具有成為cisplatin輔助藥物之潛力。

並列摘要


Cisplatin is a widely used chemotherapy drug. However, the side effects of chemotherapy such as nephrotoxicity, gastrointestinal toxicity, neurotoxicity, ototoxicity limited its used. Early studies indicated that D-methionine is sulfur-containing essential amino acid, which exerts antioxidative activity and does not affect antitumor activity. Some reports have showed that D-methionine mitigates cisplatin-induced ototoxicity. While the research of the effects D-methionine on gastrointestinal toxicity and neurotoxicity remains unknown, we examined in this study whether D-methionine can decrease cisplatin-induced side effects or not. CCK-8 assay was used to determine the cytotoxicity of D-methionine co-culture with cisplatin on human renal proximal tubular cells (HK-2 cells). The western blot and RT-PCR were applied to determine cleaved caspase 7 and AQP1 expression, respectively. In animal study, rats were divided into three groups as follows: 1) saline control group, 2) cisplatin alone group (5 mg/kg/week, intraperitoneally once a week for 3 weeks) 3) combine D-methionine (300 mg/kg/day, once a day) with cisplatin group. The results showed that chronic cisplatin-administered rats significantly decrease in body weight, food intake, WBC (white blood count), MCV (mean corpuscular volume), platelet, serum potassium concentration, uric acid and catalase activity of rat kidney. In addition, the MCHC (mean corpuscular hemoglobin concentration), serum sodium concentration, serum blood urea nitrogen (BUN) and creatinine levels, kidney/body weight ratio and renal malondialdehyde (MDA) production were increased in cisplatin-treated rats. Histological analysis revealed that large numbers of leukocytes infiltrated in proximal tubules. Vacuolization, swelling and hyaline casts were also observed in renal tubules. D-methionine administration could significantly increase body weight, food intake, MCV (mean corpuscular volume), platelet, serum potassium concentration and significantly decrease kidney/body weight ratio and renal malondialdehyde production in rat kidney. Unfortunately, higher levels of serum blood urea nitrogen and creatinine in cisplatin-treated rats could not retard through D-methionine administration. Our results indicate that D-methionine could protect against cisplatin-induced weight loss and avoid anorexia. Therefore, we suggest that D-methionine may be potential adjuvants of cisplatin.

參考文獻


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