應用化學光方法測定毛地黃毒X誘發SK-Hep-1 細胞增產之過氧化氫   全文下載

Determination of Digitoxin-induced Elevated Hydrogen Peroxide Levels in SK-Hep-1 Cells Using a Chemiluminescent Method

細胞內過氧化氫的過度增產被視為細胞凋亡的信息傳遞之上游事件。因此研發靈敏度高的過氧化氫分析法用以定量和細胞死亡相關的檢體有其重要性。吾等在此報告以魯米諾（胺基苯二醯胼）化學發光法可定量分析1 至0.1 μM（最具線性關係）之過氧化氫而該法的變異係數介於5.7% 至21.7%間。吾等假設人類肝癌細胞株SK-Hep-1 投與毛地黃毒?時可能產生過氧化氫。此項假說由毛地黃毒?處理細胞後其培養液中所含過氧化氫濃度增加和藥物劑量成正比而証實。綜合言之，因其高靈敏度，此化學發光法有潛力為定量檢體中因為毛地黃毒?服用過量中毒時引發之過氧化氫。

Drug-induced intracellular hydrogen peroxide overproduction is proposed as an upstream event in apoptosis signaling. It is therefore important to develop a sensitive method to quantify hydrogen peroxide in biological samples undergoing cell death. We report herein a luminol-based chemiluminescent assay with a suitable detection range of best linearity from 1 to 0.1 μM hydrogen peroxide with a CV range of 5.7% to 21.7% for intraday measurement at the detection range. We hypothesized that digitoxin-treated human hepatoma SK-Hep-1 cells produce hydrogen peroxide to cause cytotoxicity, which was verified by measuring the spent media by our chemiluminescent method in a dose-dependent manner. Taken together, our sensitive luminol-based chemiluminescent assay has potential use in the quantification of trace amounts of digitoxin overdose-induced hydrogen peroxide in biological samples.