Identification of ATTM as a novel H2S donor and investigation of its protective effect on HaCaT skin cells |
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Authors: | MENG Fu-hui CHEN Li XU Shi XIAN Ming ZHANG Hui LI Jian-hua DONG Qi YANG Chun-tao |
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Institution: | 1. Department of Physiology, Guangzhou Medical University, Guangzhou 511436, China;
2. Department of Chemistry, Wa-shington State University, WA Pullman 99164, USA |
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Abstract: | AIM: To investigate the ability of a metal complex ammonium tetrathiomolybdate (ATTM) to release H2S and its cytoprotective effect on an oxidative injury model. METHODS: Released H2S was absorbed in a reaction flask from ATTM dissolved in the cell medium. Staining with dichlorodihydrofluorescein diacetate or rhodamine 123 followed by photofluorography was conducted for the observation of reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) levels, respectively. Cell viability and release of lactate dehydrogenase (LDH) from the cells were measured with commercial kits. RESULTS: Similar to another H2S donor GYY4137, ATTM had an ability to release H2S in the cell medium in a dose-dependent manner. Treatment of human skin HaCaT cells with ATTM at concentrations of 25~400 μmol/L didn't significantly alter cell viability. Exposure of the cells to ultraviolet rays or a ROS donor H2O2 increased the intracellular ROS levels. Treatment with 400 μmol/L H2O2 significantly reduced the viability of HaCaT cells (P<0.01). However, before the treatment with H2O2, pretreatment with ATTM at 100 and 200 μmol/L markedly prevented the H2O2-induced cell injury (P<0.01). In addition, the treatment with H2O2 triggered ΔΨm loss (P<0.01) and LDH release from the cells (P<0.01). Prior to suffering from H2O2 injury, the preconditioning with 200 μmol/L ATTM significantly improved ΔΨm levels (P<0.05) and attenuated LDH release from the cells (P<0.01).CONCLUSION: ATTM is capable of releasing H2S and protecting human skin cells against oxidative injury. |
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Keywords: | HaCaT cells Hydrogen sulfide Ammonium tetrathiomolybdate Oxidative stress |
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