Hi all,
I want to investigate the effects of elevated ROS levels in a cell culture model. Therefore I need to scavange ROS whereas other parameters remain untouched. I already tried TEMPOL and Mito-TEMPO but both induced cell death of nearly all cells, even in otherwise untreated cells.
Does anybody know a ROS scavanger that does not influence cell viability?
As Nagendra states, toxicity is dose dependent. We have tesed N-Acetylcysteine, what also becomes toxic with higher concentration. We do research on molecular hydrogen, what is the perfect ROS scavenger. But it is difficult to keep at constant concentration, if you do not have a special incubator and a permanent H2 source.
In this publication (please see below) it is stated that sodium pyruvate, catalase, tiron and d-mannitol were non-toxic to four different cell lines. Maybe this article can give you some ideas how to set up your experiment.
Cheers, Christian
Article Responses of Solid Tumor Cells in DMEM to Reactive Oxygen Sp...
sodium pyruvate, catalase, tiron and d-mannitol also cause cell death that totally depends on dose of these chemicals. We have reported that also in our previous manuscript.
Please check our paper shown by Prof. Dr. Christian Q. Scheckhuber
As Nagendra states, toxicity is dose dependent. We have tesed N-Acetylcysteine, what also becomes toxic with higher concentration. We do research on molecular hydrogen, what is the perfect ROS scavenger. But it is difficult to keep at constant concentration, if you do not have a special incubator and a permanent H2 source.
There's a large body of literature on cell death of dopaminergic neurons of the substantia nigra pars compacta (as related to Parkinson's disease) that are attributed to ROS. That may provide some ideas for you on protective anti-ROS treatments and mechanisms. I can't remark on cell culture. But in acute rat brain slice, under the influence of complex I inhibitors, N-acetylcysteine has shown some neuroprotective effect. There are lines of inquiry into uncoupling proteins that seem to be important in a dopamine neurons ability (or inability) to handle ROS properly (see Surmeier et al, Liss and Roeper et al).
Depends on how you want your antioxidant to work. Trolox is a soluble form of vitamin E and is rather good.
N-acetylcysteine raises GSH levels and helps the whole GSH antioxidant pathways.
The EUK (-8 and 134) superoxide/catalase memitics are very good and are commercially available (I make my own and have some).
Treatment with antioxidants is only half the story, you should also always include a positive control in the form of a stressor. I typically use paraquat to increase superoxide/peroxide or add glucose oxidase to increase steady state peroxide levels.
RRR-alpha-tocopherol is non-toxic and prevents lipid peroxidation. In addition to this lipid soluble antioxidant, NAC can also be used as water-soluble antioxidant (must carefully adjust pH of stock solution to pH 7.4). Ascorbate and urate are also nontoxic water soluble antioxidants.
In my hand both SOD and NAC worked fine. I have tried many others but they showed the best effect. We need to determine the stress level as mitochondria and ER can induce ROS. NAC can reduce both of them. Thanks, and good luck in your research.
Thanks to all of you, for your answers. I now tried trolox in my cell culture and got quite good results. So thanks for the recommendation.
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