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DIDS protects against neuronal injury by blocking Toll-like receptor 2 activated-mechanisms.

ABSTRACT: Using an in vitro ischemia model (ischemic solution; IS model) that induces penumbral cell death, we examined the effect of 4,4'-diisothio-cyanostilbene-2,2'-disulfonic acid (DIDS) on cell injury/death and underlying molecular mechanisms. Propidium iodide (PI) uptake was used to quantify cell death in organotypic hippocampal slice cultures. A 24-h IS exposure caused a fivefold increase in mean PI fluorescence intensity. DIDS, dose-dependently (1-4000 microM), reduced the IS-induced PI uptake in hippocampal CA1 neurons with an IC(50) of 26 microM. This protective effect of DIDS was reversible and effective even 6 h following the onset of IS treatment. Gene expression profiling studies indicated that among approximately 46,000 transcripts tested, the most significantly up-regulated gene by IS was interleukin-1beta (IL-1beta) which was also the most significantly down-regulated gene when DIDS was added to the IS-treated slices. The addition of a recombinant IL-1 receptor antagonist (100 microg/mL) or neutralizing IL-1beta antibody significantly attenuated the IS-induced cell death, indicating that the up-regulation of IL-1beta with IS treatment contributed to the IS-induced cell death. Toll-like receptor 2 (TLR2), another gene that was significantly up-regulated by IS and suppressed by DIDS, was studied to determine whether it was related to the IL-1beta up-regulation. Indeed, this was the case as the IS-induced IL-1beta up-regulation was abolished in TLR2-/- mouse brain slices. Furthermore, the IS-induced cell death was significantly reduced in TLR2-/- when compared with that in wild-type slices, indicating that TLR2 is functionally upstream of IL-1beta in this IS model. We conclude that (i) IS up-regulates TLR2 expression and augments TLR2 signaling, causing over-expression of IL-1beta which leads to cell death and (ii) DIDS blocks IS-induced neuronal injury, at least partially, by suppressing the TLR2 pathway.

SUBMITTER: Yao H

PROVIDER: S-EPMC2782475 | biostudies-literature | 2009 Feb

REPOSITORIES: biostudies-literature

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DIDS protects against neuronal injury by blocking Toll-like receptor 2 activated-mechanisms.

Yao Hang H Felfly Hady H Wang Juan J Zhou Dan D Haddad Gabriel G GG

Journal of neurochemistry 20081210 3

Using an in vitro ischemia model (ischemic solution; IS model) that induces penumbral cell death, we examined the effect of 4,4'-diisothio-cyanostilbene-2,2'-disulfonic acid (DIDS) on cell injury/death and underlying molecular mechanisms. Propidium iodide (PI) uptake was used to quantify cell death in organotypic hippocampal slice cultures. A 24-h IS exposure caused a fivefold increase in mean PI fluorescence intensity. DIDS, dose-dependently (1-4000 microM), reduced the IS-induced PI uptake in ...[more]

PMID: 19077053

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Project description:Peroxisome proliferator-activated receptor-? is significantly down-regulated in circulating leukocytes from children with sepsis. Peroxisome proliferator-activated receptor-? null (Ppara) mice have greater mortality than wild-type mice when subjected to sepsis by cecal ligation and puncture. We sought to characterize the role of peroxisome proliferator-activated receptor-? in sepsis and to identify the mechanism whereby peroxisome proliferator-activated receptor-? confers a survival advantage.Prospective randomized preclinical study.Laboratory investigation.Male C57Bl/6J and Ppara mice (B6.129S4-Ppara/J), aged 12-16 weeks.Bone marrow chimeric mice were generated and subjected to cecal ligation and puncture. Survival was measured for 7 days. Separate groups of nontransplanted mice underwent cecal ligation and puncture and were euthanized 24 hours later for plasma and tissue analyses.Ppara mice had dramatically reduced survival compared with wild-type mice irrespective of the peroxisome proliferator-activated receptor-? status of the bone marrow they received (3% vs 63%; p < 0.0001). No difference in survival was observed between Ppara mice that received wild-type versus Ppara marrow or in wild-type mice receiving wild-type versus Ppara marrow. In septic, nontransplanted mice at 24 hours, Ppara mice had elevated cardiac troponin levels compared with wild-type mice. Cardiac histologic injury scores were greater in Ppara versus wild-type mice. Expression of transcription factors and enzymes related to fatty acid oxidation in the heart were profoundly down-regulated in both wild-type and Ppara mice, but more so in the Ppara mice.Peroxisome proliferator-activated receptor-? expression in nonhematopoietic tissues plays a critical role in determining clinical outcome in experimental polymicrobial sepsis and is more important to survival in sepsis than hematopoietic peroxisome proliferator-activated receptor-? expression. Cardiac injury due to inadequate energy production from fatty acid substrate is a probable mechanism of decreased survival in Ppara mice. These results suggest that altered peroxisome proliferator-activated receptor-?-mediated cellular metabolism may play an important role in sepsis-related end-organ injury and dysfunction, especially in the heart.

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DIDS protects against neuronal injury by blocking Toll-like receptor 2 activated-mechanisms.

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DIDS protects against neuronal injury by blocking Toll-like receptor 2 activated-mechanisms.

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