i want to induce hypoxia invivo (RAT or MICE), to check its effect on nrf2 mediated regulation of some target proteins in heart. and i want to use atleast two methods to induce hypoxia to get confirmed and stable results. can anyone help me with the exact protocol for this? for chemically induced hypoxia , please also give me the dose/concentration of reagent/chemical used to induce hypoxia.
any help and guidance are highly appreciated. Thanks
The redox sensitive transcription factor Nrf2 is activated by hypoxia/reoxygenation (whole body in vivo exposure) or local ischemia/reperfusion injury through temporary occlusion of blood flow to an organ under anaesthesia. The times required for each of these procedures varies dependent on the organ of interest. However, there is plenty of literature on this if you use Nrf2 and hypoxia as key words in Pubmed. The following paper pertains to the brain, but with whole animal exposure to hypoxia various organs are likely to show activation of Nrf2. These authors report on the chemical activation of Nrf2 as well.
https://www.ncbi.nlm.nih.gov/pubmed/28298215
Good luck with your experiments.
thank you for your response Pearson.
i want to induce hypoxia in heart. one option is using hypoxic chambers with (6% O2, etc), other is LCA ligation,to mimic ischemia/hypoxia. i was interested if there is some dug/chemical that could induce hypoxia in heart, what concentration should i use for that?
plus i want to know that if the method is used to induce hypoxia in brain, can we use the same conditions ( i.e 6%O2 for 1hr or concentration of a reagent ) to induce hypoxia in heart?
The link to the paper on brain hypoxia refers to a drug that can stimulate the whole body if delivered systemically. Naturally, exposing rats to chronic hypoxia gas also makes the heart hypoxic. Note that for mice brief exposure to 6-8% oxygen is better tolerated than in rats, so gas levels may need to be titrated with trial and error.
you have mentioned that ''hypoxia activate nrf2'' but some papers show the opposite,
https://www.readbyqxmd.com/read/28128361/protective-effect-of-butin-against-ischemia-reperfusion-induced-myocardial-injury-in-diabetic-mice-involvement-of-the-ampk-gsk-3%C3%AE-nrf2-signaling-pathway
what should i follow now?
I am sorry but my reading of this paper is not the opposite, Nrf2 is a redundant part of a complicated antioxidant pathway. Of course there are multiple pathways involved, but if you examine added factors such as hyperglycemia it naturally becomes more complicated. Nonetheless, the role has not really changed.
I suggest that you review all the known effects/roles of Nrf2 in antixodant pathways to understand the subtle differences between studies, However, in my mind these studies do not contradict each other. More study of these mechanisms would help you to understand this.
sorry i shared the wrong paper. i read some other papers and that had confused me so much. now i have understood it. thanks again for your worthy explanation and help.
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