人參皂苷Rg1與N-乙醯半胱胺酸對於順氯氨鉑引發腎毒性之純系小鼠腎組織中甲基乙二醛及D-乳酸的影響

Effects of ginsenoside Rg1 and N-acetylcysteine on methylglyoxal and D-lactate in cisplatin-induced nephrotoxicity kidney tissues in inbred mice

蔡牧承(Mu-Cheng Tsai)

指導教授 ： 林本元

臺北醫學大學/藥學院/藥學系(碩博士班)/碩士(2014年)

摘要

本研究利用順氯氨鉑(CDDP)誘導腎毒性模型，偵測使用人參皂苷Rg1 (ginsenoside Rg1, GS Rg1)及N-乙醯半胱胺酸(N-acetylcysteine, NAC)後腎組織中甲基乙二醛(MG)與D-乳酸之濃度，以評估MG及D-乳酸作為組織損傷指標之可能性。實驗使用30隻六週齡雌性BALB/c小鼠隨機分為五組。除Normal組外，其餘四組在第六天利用腹腔注射CDDP 5 mg/kg/day誘導腎損傷。此外，自第一天起連續十天分別投與NAC 450 mg/kg/day、GS Rg1 5 mg/kg/day及共同投與兩藥物，Normal與Control則給與0.1 mL之d.d. water作為對照。動物犧牲後，取其腎組織均質並進行螢光衍生化處理再利用螢光高效液相層析進行分析。實驗結果顯示Control組MG和D-乳酸含量相較Normal組有顯著增加(MG：64.6 ± 22.0 vs. 284.1 ± 327.3 μg/mg, p < 0.01；D-乳酸：11.22 ± 1.52 vs. 20.74 ± 16.21 μmol/g, p < 0.01)。而在給與GS Rg1、NAC或GS Rg1 + NAC其腎組織中MG和D-乳酸均有下降的趨勢，並且在GS Rg1 + NAC組別D-乳酸顯著降低到10.96 ± 2.85 μmol/g。此外，在組織學切片的觀察發現損傷程度下降的趨勢與組織中的D-乳酸濃度下降趨勢具有良好的相關性。因此，我們認為組織中D-乳酸之含量可反應出腎損傷的程度，並且D-乳酸相較MG可做為組織損傷的指標。

關鍵字

D-乳酸 ； 甲基乙二醛 ； 人參皂苷Rg1 ； N-乙醯半胱胺酸 ； 順氯氨鉑誘導之腎毒性

並列摘要

In this study, we investigated the effect of ginsenoside Rg1 (GS Rg1) and N-acetylcysteine (NAC) on the concentration of methylglyoxal (MG) and D-lactate and evaluated the MG and D-lactate as indicators in cisplatin-induced nephrotoxicity kidney tissues.
We divided 30 six-week-old female BALB/c mice into five groups. Then, we used cisplatin (5 mg/kg/day) intraperitoneally to induce nephrotoxicity for 5 day except for the Normal group at the 6th day. Besides, the mice were orally administered GS Rg1 5 mg/kg/day, NAC 450 mg/kg/day and GS Rg1+NAC for 10 days, respectively. At the 10th day, the mice were sacrificed and the kidney tissue was homogenized and centrifuged. The supernatants were derivatized with fluorescent reagent 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) or 5,6-diamino-2,4-
dihydroxypyrimidine (DDP) for D-lactate and MG determination.
The results showed that the levels of MG and D-lactate increased significantly in the Control group compared with the Normal group. (MG: 64.6 ± 22.0 vs. 284.1 ± 327.3 μg/mg, D-lactate: 11.22 ± 1.52 vs. 20.74 ± 16.21μmol/g, p < 0.01) Administration of GS Rg1, NAC or GS Rg1+NAC revealed a decreasing tendency of both the levels of MG and D-lactate. Particularly, the D-lactate level significantly decreased to 10.96 ± 2.85 μmol/g in the GS Rg1+NAC group. In addition, it had good co-relationship between the D-lactate level and the histological outcome. As a result, we considered that the D-lactate level reflects the tissue injuries and the D-lactate level is a better indicator to tissue injuries than MG.

並列關鍵字

D-lactate ； methylglyoxal ； ginsenoside Rg1 ； N-acetylcysteine ； cisplatin-induced nephrotoxicity

參考文獻

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