Targeting thioredoxin-1 with siRNA exacerbates oxidative stress injury after cerebral ischemia/reperfusion in rats

Neuroscience. 2015 Jan 22:284:815-823. doi: 10.1016/j.neuroscience.2014.10.066. Epub 2014 Nov 8.

Abstract

Reactive oxygen species and their detrimental effects on the brain after transient ischemia/reperfusion (I/R) have been implicated in the pathogenesis of ischemic reperfusion injury. Thioredoxin-1 (Trx-1) is an endogenous antioxidant protein that has neuroprotective effects. We hypothesized that Trx-1 plays a crucial role in regulating cerebral I/R injury. To be able to test this, 190 Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion (tMCAO) with Trx-1 siRNA (small interference RNA) injected 24 h prior to ischemia. At 24 h after tMCAO, we measured neurological deficits, infarct volume, and brain water content, and found that neurological dysfunction, brain infarct size, and brain edema were worse in the Trx-1 siRNA group than in the control group. Oxidative stress was evaluated by measuring superoxide dismutase activity and malondialdehyde level. The levels of Trx-1 and its cofactor, peroxiredoxin (Prdx), were significantly decreased after Trx-1 down-regulated. However, there is no significant difference in the Prdx mRNA level after administration of Trx-1 siRNA. In contrast, Prdx-SO3 protein levels were significantly increased in the Trx-1 siRNA group. We also investigated the specific role of nuclear factor erythroid 2-related factor 2 (Nrf2) in Trx-1 induction by knocking down Nrf2. Nrf2 siRNA injection decreased Trx-1 mRNA and protein expression. Our results suggest that the exacerbation of brain damage was associated with enhanced cerebral peroxidation in brain tissues. Moreover, these results revealed that Trx-1, which is more likely regulated by Nrf2, exerts a neuroprotective role probably through maintaining the reduction activity of Prdx1-4.

Keywords: nuclear factor erythroid 2-related factor 2; oxidative stress; peroxiredoxin; thioredoxin-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / pathology
  • Brain / physiopathology*
  • Brain Edema / pathology
  • Brain Edema / physiopathology
  • Brain Ischemia / pathology
  • Brain Ischemia / physiopathology*
  • Disease Models, Animal
  • Gene Knockdown Techniques
  • Infarction, Middle Cerebral Artery
  • Male
  • Malondialdehyde / metabolism
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidative Stress / physiology*
  • Peroxiredoxins / metabolism
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / administration & dosage
  • Random Allocation
  • Rats, Sprague-Dawley
  • Reperfusion Injury / pathology
  • Reperfusion Injury / physiopathology*
  • Superoxide Dismutase / metabolism
  • Thioredoxins / genetics
  • Thioredoxins / metabolism*

Substances

  • NF-E2-Related Factor 2
  • Nfe2l2 protein, rat
  • RNA, Messenger
  • RNA, Small Interfering
  • Txn1 protein, rat
  • Malondialdehyde
  • Thioredoxins
  • Peroxiredoxins
  • Superoxide Dismutase