Tyloxapol被報導可以用來預防巨噬細胞受內毒素引發刺激活化作用，但對於Tyloxapol與巨噬細胞所產生的細胞內反應卻沒有完整的研究，因此在我們的研究中將探討巨噬細胞與tyloxapol作用後，所引發的各種細胞內反應，以及tyloxapol對受內毒素刺激活化之巨噬細胞的抗氧化作用。我們利用流式細胞儀來測定細胞內之變化，包括：ROS的含量、粒線體膜電位的變化、細胞大小與複雜度、及分析細胞週期，同時也偵測tyloxapol對由內毒素引發活化之巨噬細胞所產生的抗氧化作用。結果顯示細胞內H2O2含量，隨著tyloxapol劑量增加而下降，而細胞內O2−含量隨著tyloxapol劑量增加而漸漸上升，另外tyloxapol也會使粒線體膜電位呈現不穩定狀態，在tyloxapol為高劑量且培養時間為4小時，tyloxapol更會引發細胞產生凋亡作用。當tyloxapol為低劑量且培養時間小於2小時，tyloxapol才能對細胞產生抗氧化作用。在tyloxapol為低劑量時，巨噬細胞內O2−及H2O2之變化量可回復至與沒有加入tyloxapol之巨噬細胞相當狀態。我們同時也研究加入tyloxapol之Hela子宮頸癌細胞之細胞內各種變化情形，以便了解tyloxapol產生之效應是否僅針對特定細胞。我們的研究可促使更了解tyloxapol界面活性劑對各種細胞之分子作用機轉。 Tyloxapol is reported to prevent macrophages from reacting to endotoxin. However, the intracellular responses that tyloxapol induces in macrophages are still not fully explored. Hence, the objective of this study was to evaluate the intracellular events in macrophages treated with tyloxapol and assess the antioxidant properties of tyloxapol in endotoxin-activated macrophages. Using flow cytometry, we examined intracellular responses in macrophages: reactive oxygen species (ROS) content, mitochondria membrane potential, and cell cycle profiles.We also assessed the antioxidant properties of tyloxapol in endotoxin-activated macrophages. Kinetic hydrogen peroxide production tended to decline with increasing doses. Tyloxapol produced a progressive increase followed by a decline in superoxide anion production in macrophages with increasing doses. Tyloxapol also caused unstable fluctuations in mitochondrial membrane potential. Apoptosis had developed at higher doses after 4 h of incubation time. After 2 h of tyloxapol-pretreatment, tyloxapol acted as an antioxidant only at lower doses. Most tyloxapol-pretreated cells at lower doses fully recovered from the changes in superoxide anion and hydrogen peroxide production. We also investigate the intracellular events in Hela cell to evaluate cell-dependent properties induced by tyloxapol.Our findings contribute to a better understanding of the molecular action of tyloxapol in various cells such as macrophages and cancer cells.
