- 學位論文
人參與水楊酸鈉在順氯氨鉑引發之腎毒性於純系小鼠的藥效評估及人參成分的含量分析
Effects of ginseng and sodium salicylate on cisplatin-induced nephrotoxicity in inbred mice and quantitative analysis of ginsenosides in ginseng extract
摘要
順氯氨鉑 (cis-diamminedichloroplatinum (Ⅱ), CDDP) 是目前廣泛用於治療固體癌的化學治療藥物之一。此藥物的藥效良好，然而藥物引起的腎毒性卻成為臨床上限制其使用的主要原因。本研究目的即為評估人參及水楊酸鈉 (sodium salicylate, S) 於CDDP所引起之腎炎的預防效果，並進一步利用高壓液相層析法 (high performance liquid chromatography, HPLC) 定量所使用之人參中人參皂苷的含量。 實驗以六週齡雌鼠 (BALB/c mice) 為實驗動物，採取腹腔內注射方式連續五天給予CDDP 5 mg/kg/d以引發腎炎。在給予CDDP的前五天開始經口投予人參濃縮劑 (ginseng extract, GE) 250 mg/kg/d、S 100 mg/kg/d或合併GE+S作為預防藥物。在人參含量分析方面，實驗選用Inertsil ODS-2 column (4.6x150 mm, particle size 5 μm)，移動相為acetonitrile : H2O = 20 : 80 (ginsenoside Rg1) 及acetonitrile : H2O = 30 : 70 (ginsenoside Rb1與Rd)，測定波長為203 nm。 實驗結果顯示GE及S對於尿中N-acetyl-β-D-glucosaminidase (NAG)、肌酸酐 (urine creatinine)、尿蛋白 (urine protein) 與血中尿素氮 (BUN) 皆有不同程度的改善效果；腎組織損傷相較於對照組也有減緩的趨勢。在免疫螢光染色方面，TNF-α (tumor necrosis factor-α) 的量明顯減少，而p21及PCNA (proliferating cell nuclear antigen) 的表現則有不同程度的增加。在含量分析部份，實驗中使用之人參濃縮劑中人參皂苷含量經HPLC定量結果依序為Rb1＞Rg1＞Rd。 綜合實驗結果可推論，人參濃縮劑及水楊酸鈉透過其抑制發炎反應，阻止細胞週期的進展並促進DNA修復等作用，因此具有腎臟保護的效果。
並列摘要
Cisplatin (cis-diamminedichloroplatinum (Ⅱ), CDDP) is one of the most commonly used antineoplastic agents in the treatment of various solid tumors. However, the full clinical utility of CDDP is limited because of its dose-related nephrotoxic side effects. The purpose of this study was to evaluate the preventive effects of ginseng extract (GE) and sodium salicylate (S) on CDDP-induced nephrotoxicity in bred mice. Besides, this study also did quantitative analysis by high performance liquid chromatography (HPLC) to comfirm the quantity of ginsenosides in GE. In this study, six-week-old female BALB/c mice were administered with 5 mg/kg/day of CDDP intraperitoneally for 5 days. Preventive drugs including 250 mg/kg of GE, 100 mg/kg of sodium salicylate, and combination of GE and S were given orally once daily from 5 days before CDDP administration respectively. Besides, in this quantitative analysis, ginsenosides have been separated and identified on an Inertsil ODS-2 column (4.6 x 150 mm, particle size 5 μm) with elution using acetonitrate and water (acetonitrile：H2O = 20 : 80 for ginsenoside Rg1；acetonitrile : H2O = 30 : 70 for ginsenoside Rb1 and Rd) as the mobile phase. The temperature was maintained room temperature, and detection wavelength was 203 nm. The treatment groups showed improvements in urine N-acetyl-β-D-glucosaminidase (NAG), urine creatinine excretion, urine protein and blood urea nitrogen (BUN) at different levels. Furthermore, the treatment groups ameliorated CDDP-induced renal morphological damages, diminished TNF-α deposited in injury tissues, and increased the expression of p21 and PCNA in renal cells as well. The result of HPLC revealed that GE contained more ginsenoside Rb1 than Rg1 and Rd. Our findings demonstrated that GE and S attenuate CDDP-induced nephrotoxicity probably by inhibiting TNF-α expression and promoting cell cycle arrest to repair DNA damage.