馬兜鈴酸是一種由植物本身所產生的一種致癌毒素，主要常見於中草藥及一些減肥食品中，馬兜鈴酸的毒性不只引起人類的腎臟病變，亦會導致泌尿系統的癌症產生。國際上已公認它是一種致癌物，我們欲藉由HL-60細胞株(人類前骨髓性白血病細胞株)來探討馬兜鈴酸所造成的細胞毒性以及基因毒性之分子機制。我們藉由MTT assay觀察到以不同濃度的馬兜鈴酸處理HL-60細胞株24小時，細胞的存活率會隨濃度的增加而存活率相對下降。若將HL-60細胞株曝露在馬兜鈴酸下處理一小時，以DCFH-DA作為分子探針可偵測到有ROS的產生，並且若經過ROS抑制劑處理，馬兜鈴酸所誘發的ROS含量即有下降的趨勢。而抽取HL-60細胞株全蛋白來探討馬兜鈴酸與p38磷酸化的關係，則發現p38 的活化會隨著馬兜鈴酸所處理的時間以及劑量的增加，而磷酸化程度也與之遽增。此外，若將ROS抑制劑與馬兜鈴酸共同處理細胞，則可以發現p38蛋白磷酸化有受到抑制的現象。根據以上發現，我們認為馬兜鈴酸所導致之ROS以及MAPK pathway的產生與馬兜鈴酸的基因毒性及細胞毒性
Aristolochic acid (AA) is a carcinogen which produced from the Aristolochia spp.. It is found primarily in Chinese herbals and slimming remedies. Aristolochic acid has been associated with the development of a novel nephropathy, designated aristolochic acid nephropathy (AAN), and urothelial cancer in AAN patients. Treatment of various concentration of AA from 25 to 200 μM, a significant decrease to 60% in cell viability was observed when HL-60 cultures were incubated with AA for 24 h at a concentration of 200 μM. AA induces Reactive oxygen species (ROS) was also detected using DCFH-DA as a probe in HL-60. Western blotting showed that AA can activate the protein phosphorylation╱dephosphorylation in HL-60 cell. The results showed that treatment of various concentration of AA induces in a dose-dependent increase in p38 kinase phosphorylation. Treatment of HL-60 with ROS inhibitor and AA, a p38 protein kinase phosphorylation was reduced. According to our results, AA induced ROS and p38 protein kinase phosphorylation was associated with its genotoxicity and cytotoxicity.
