Zinc-dependent and independent actions of hydroxyhydroquinone on rat thymic lymphocytes

Coffee contains hydroxyhydroquinone (HHQ). HHQ is one of the by-products released during bean roasting. Therefore, it is important to elucidate the bioactivity of HHQ to predict its beneficial or adverse effects on humans. We studied zinc-dependent and independent actions of commercially procured synthetic HHQ in rat thymocytes using flow cytometric techniques with propidium iodide, FluoZin-3-AM, 5-chloromethylfluorescein diacetate, and annexin V-FITC. HHQ at 1050 µM elevated intracellular Zn²⁺ levels by releasing intracellular Zn²⁺. HHQ at 10 µM increased cellular thiol content in a zinc-dependent manner. However, HHQ at 30-50 µM reduced cellular thiol content. Although the latter actions of HHQ (30-50 µM) were suggested to increase cell vulnerability to oxidative stress, HHQ at 0.3-100 µM significantly protected cells against oxidative stress induced by H₂O₂. The process of cell death induced by H₂O₂ was delayed by HHQ, although both H₂O₂ and HHQ increased the population of annexin V-positive living cells. However, HHQ at 10-30 µM promoted cell death induced by A23187, a calcium ionophore. HHQ at 10-30 µM exerted contrasting effects on cell death caused by oxidative stress and Ca²⁺ overload. Because HHQ is considered to possess diverse cellular actions, coffee with reduced amount of HHQ may be preferable to avoid potential adverse effects.