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Synthetic Oligodeoxynucleotides Containing Multiple Telemeric TTAGGG Motifs Suppress Inflammasome Activity in Macrophages Subjected to Oxygen and Glucose Deprivation and Reduce Ischemic Brain Injury in Stroke-Prone Spontaneously Hypertensive Rats.

Zhao J, Mou Y, Bernstock JD, Klimanis D, Wang S, Spatz M, Maric D, Johnson K, Klinman DM, Li X, Li X, Hallenbeck JM - PLoS ONE (2015)

Bottom Line: Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells.In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion.Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Jinan Central Hospital affiliated with Shandong University, 105 Jiefang Road, Jinan, Shandong, 250013, P. R. China; Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT
The immune system plays a fundamental role in both the development and pathobiology of stroke. Inflammasomes are multiprotein complexes that have come to be recognized as critical players in the inflammation that ultimately contributes to stroke severity. Inflammasomes recognize microbial and host-derived danger signals and activate caspase-1, which in turn controls the production of the pro-inflammatory cytokine IL-1β. We have shown that A151, a synthetic oligodeoxynucleotide containing multiple telemeric TTAGGG motifs, reduces IL-1β production by activated bone marrow derived macrophages that have been subjected to oxygen-glucose deprivation and LPS stimulation. Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells. In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion. These findings clearly suggest that the modulation of inflammasome activity via A151 may contribute to a reduction in pro-inflammatory cytokine production by macrophages subjected to conditions that model brain ischemia and modulate ischemic brain damage in an animal model of stroke. Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.

No MeSH data available.


Related in: MedlinePlus

A151 reduced IL-1β and caspase 1 maturation, and the expression of NLRP3 and iNOS in BMDM subjected to LPS and OGD.(A) A151 reduced mature IL-1β in supernatants. (B) A151 reduced mature caspase 1 in supernatants. (C) A151 did not influence caspase 11 in cell lysates or supernatants. (D) A151 reduced NLRP3 in cell lysates. (E) A151 reduced iNOS in cell lysates. Data are presented as mean ± SEM from three replicates representative of three independent experiments (**, p < 0.05 compared with control or C151 treatment).
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pone.0140772.g002: A151 reduced IL-1β and caspase 1 maturation, and the expression of NLRP3 and iNOS in BMDM subjected to LPS and OGD.(A) A151 reduced mature IL-1β in supernatants. (B) A151 reduced mature caspase 1 in supernatants. (C) A151 did not influence caspase 11 in cell lysates or supernatants. (D) A151 reduced NLRP3 in cell lysates. (E) A151 reduced iNOS in cell lysates. Data are presented as mean ± SEM from three replicates representative of three independent experiments (**, p < 0.05 compared with control or C151 treatment).

Mentions: The effect of A151 on IL-1β expression and maturation was explored further by western blot analysis. A151 treatment of oxygen and glucose deprived BMDM reduced the levels of mature IL-1β in cell culture supernatants (Fig 2A). Being that the inflammasome is a multiprotein complex and a key regulator of IL-1β production, we studied the regulatory potential of A151 on the expression/protein levels of additional inflammasome components. A151 reduced mature caspase-1 (Fig 2B) and NLRP3 (Fig 2D), but did not affect ASC, AIM2, NLRP1 or NLRC4 (data not shown). Further it is known that IL-1β can induce the expression of iNOS [48] and that iNOS can influence stroke-induced cellular damage [49]. We therefore analyzed iNOS levels and found that A151 reduced the levels of iNOS expression (Fig 2E). It is important to note that in murine systems caspase-11 in part controls IL-1β secretion via the potentiation of caspase-1 activation and can thus induce caspase-1-independent pyroptosis downstream of non-canonical NLRP3 inflammasome activators. We thus sought to check the expression and maturation of caspase-11. Of note, A151 did not influence the levels of pro- and/or mature caspase-11 in cell lysates or supernatants (Fig 2C).


Synthetic Oligodeoxynucleotides Containing Multiple Telemeric TTAGGG Motifs Suppress Inflammasome Activity in Macrophages Subjected to Oxygen and Glucose Deprivation and Reduce Ischemic Brain Injury in Stroke-Prone Spontaneously Hypertensive Rats.

Zhao J, Mou Y, Bernstock JD, Klimanis D, Wang S, Spatz M, Maric D, Johnson K, Klinman DM, Li X, Li X, Hallenbeck JM - PLoS ONE (2015)

A151 reduced IL-1β and caspase 1 maturation, and the expression of NLRP3 and iNOS in BMDM subjected to LPS and OGD.(A) A151 reduced mature IL-1β in supernatants. (B) A151 reduced mature caspase 1 in supernatants. (C) A151 did not influence caspase 11 in cell lysates or supernatants. (D) A151 reduced NLRP3 in cell lysates. (E) A151 reduced iNOS in cell lysates. Data are presented as mean ± SEM from three replicates representative of three independent experiments (**, p < 0.05 compared with control or C151 treatment).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4608557&req=5

pone.0140772.g002: A151 reduced IL-1β and caspase 1 maturation, and the expression of NLRP3 and iNOS in BMDM subjected to LPS and OGD.(A) A151 reduced mature IL-1β in supernatants. (B) A151 reduced mature caspase 1 in supernatants. (C) A151 did not influence caspase 11 in cell lysates or supernatants. (D) A151 reduced NLRP3 in cell lysates. (E) A151 reduced iNOS in cell lysates. Data are presented as mean ± SEM from three replicates representative of three independent experiments (**, p < 0.05 compared with control or C151 treatment).
Mentions: The effect of A151 on IL-1β expression and maturation was explored further by western blot analysis. A151 treatment of oxygen and glucose deprived BMDM reduced the levels of mature IL-1β in cell culture supernatants (Fig 2A). Being that the inflammasome is a multiprotein complex and a key regulator of IL-1β production, we studied the regulatory potential of A151 on the expression/protein levels of additional inflammasome components. A151 reduced mature caspase-1 (Fig 2B) and NLRP3 (Fig 2D), but did not affect ASC, AIM2, NLRP1 or NLRC4 (data not shown). Further it is known that IL-1β can induce the expression of iNOS [48] and that iNOS can influence stroke-induced cellular damage [49]. We therefore analyzed iNOS levels and found that A151 reduced the levels of iNOS expression (Fig 2E). It is important to note that in murine systems caspase-11 in part controls IL-1β secretion via the potentiation of caspase-1 activation and can thus induce caspase-1-independent pyroptosis downstream of non-canonical NLRP3 inflammasome activators. We thus sought to check the expression and maturation of caspase-11. Of note, A151 did not influence the levels of pro- and/or mature caspase-11 in cell lysates or supernatants (Fig 2C).

Bottom Line: Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells.In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion.Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Jinan Central Hospital affiliated with Shandong University, 105 Jiefang Road, Jinan, Shandong, 250013, P. R. China; Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT
The immune system plays a fundamental role in both the development and pathobiology of stroke. Inflammasomes are multiprotein complexes that have come to be recognized as critical players in the inflammation that ultimately contributes to stroke severity. Inflammasomes recognize microbial and host-derived danger signals and activate caspase-1, which in turn controls the production of the pro-inflammatory cytokine IL-1β. We have shown that A151, a synthetic oligodeoxynucleotide containing multiple telemeric TTAGGG motifs, reduces IL-1β production by activated bone marrow derived macrophages that have been subjected to oxygen-glucose deprivation and LPS stimulation. Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells. In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion. These findings clearly suggest that the modulation of inflammasome activity via A151 may contribute to a reduction in pro-inflammatory cytokine production by macrophages subjected to conditions that model brain ischemia and modulate ischemic brain damage in an animal model of stroke. Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.

No MeSH data available.


Related in: MedlinePlus